TO STUDY THE ANTIOXIDANT AND ANTI-INFLAMMATORY PROPERTIES OF OIL FROM LEAVES OF CAMELLIA SINENSIS L. IN EXPERIMENTAL ANIMAL MODELS

TO STUDY THE ANTIOXIDANT AND ANTI-INFLAMMATORY PROPERTIES OF OIL FROM LEAVES OF CAMELLIA SINENSIS L. IN EXPERIMENTAL ANIMAL MODELS

Meghamala Mandal, Shrabanti Pyne, Supriya Bhowmik, Deblina Giri, Koushik Das * and Jayasree Laha

Department of Nutrition, Belda College, Belda, Paschim Medinipur, West Bengal, India.

ABSTRACT: The present study has been conducted to search out the anti-oxidative and anti-inflammatory potential of leaves of Camellia sinensis L. oil (Tea oil, TO).  Forty-eight rats were included for 14 days experiment namely: Group I was normal to control, Group II was vehicle control, and Group III to VIII rats were subjected to paracetamol at the dose of 15 mg/ kg body weight/day to induce oxidative stress and inflammation (OS and I rats). Group IV to VII rats orally received TO at the dose of 100, 200, 400, 800 mg/kg, respectively and Group VIII received olive oil at 400 mg/kg. Paracetamol causes a significant (p < 0.05) increase in plasma urea, creatinine, glutamine-oxalic-transaminase, glutamate-pyruvate-transaminase, alkaline-phosphatase, interleukin-18 and tissue-injury-molecule-1, malondialdehyde and degeneration of hepatic cells in case of only OS and I rats. But treatment with TO in different doses showed a significant (p < 0.05) decrease above said parameters. We observed a significant (p < 0.05) decrease in superoxide dismutase, catalase, interleukin-10 levels in OS and I rats and also treatment with TO showed a significant (p < 0.05) increase in superoxide dismutase, catalase, interleukin-10 level in group IV to VII compared with only group III rats. Tea oil shows potent antioxidative activities against high-dose paracetamol-induced oxidative stress and inflammation by regulating the levels of pro-inflammatory and anti-inflammatory cytokines, whereas the most effective dose TO was 400 mg/kg.

Keywords: Inflammation, Oxidative stress, Paracetamol, Tissue injury molecule, C-reactive protein

INTRODUCTION: In our previous study, Camellia sinensis L. oil (Tea oil or TO) was used for the determination of fatty acid profile, anticancer activity using MCF-7 cell lines, antimicrobial activities, and antioxidative activities in-vitro.

When we found that TO contains mostly linolenic omega-3 fatty acid and it has remarkable anticancer potential, antimicrobial and antioxidative potential ¹. In the present study, an approach has been followed to determine the anti-oxidative and anti-inflammatory potential of TO on an experimental animal model.

Oxidative stress plays a pivotal role in excessive production of cytokines, producing tissue injury molecules (TIM), and thus leads to inflammation due to tissue injuries and instigating lipid peroxidation ². If oxidative stress and inflammation remain unattended at this stage, it may lead to certain diseases such as atherosclerosis, diabetes, heart disease, liver and kidney disease even cancer ^{3, 4}. On the other hand, inflammatory cells start liberating various types of reactive species like superoxide, and hydrogen peroxide, which lead to exaggerated oxidative stress. At the same time, several reactive species may initiate intracellular signaling for increasing pro-inflammatory cytokines. Thus it can be said that oxidative stress and inflammation are closely interlinked with pathophysiological events ⁵. Various studies have been published so far show the anti-oxidative and anti-inflammatory effect of plant oil anti-oxidative and anti-inflammatory effects of plant oil against paracetamol (N- acetyl- para- aminophenol, APAP) induced oxidative stress and inflammation ⁶. Lavender oil (Lavendulla angustifolia) ², tea tree oil (Melaleuca alternifolia) ⁷, Trachydium royeli plant oil ⁴ and hinoki oil (Chamaecyparis obtuse) ⁸ possess a strong anti-oxidative and anti-inflammatory activities. Treatment with ethyl acetate fraction of methanolic extract of Camellia sinensis on streptozotocin-induced diabetic rats could significantly recover oxidative stress ⁹. The present study is based on the fact that no experimental animal studies on the therapeutic efficacy of TO extract obtained from Camellia sinensis L. have been performed by any researcher till today. For this achievement, the research method has been designed to evaluate the anti-oxidative and anti-inflammatory potential of TO on oxidative stress and inflammation (OS and I) created by inducing high dose APAP in the rat model.

MATERIALS AND METHODS:

Collection of Tea Leaves and Preparation of Tea Oil (TO): Fresh tea leaves were collected from heritage tea garden, Gopali, Indian Institute of Technology, Kharagpur, (Geographical extension between 22 °28' 59" North to 22 °29' 19" North latitude and 87 °30' 20" East to 87 °30' 35" East), Paschim Medinipur District of West Bengal, India. The leaves were identified by a taxonomist (CNH/28/2018), and the voucher specimen was deposited in the Department of Botany, Raja Narendra Lal Khan Women's College (Autonomous) for future reference. The oil content was extracted from Camellia sinensis L. leaves by the Soxhlet apparatus using the solvent extraction method according to the American oil chemists’ society (AOCS) method. At first, 1000g Camellia sinensis L. leaves were soaked in warm water at 60ºC for 5 minutes and discarded the water to remove the water-solving compounds from the tea leaves. Then the wet tea leaves were dried at 40 ± 1°C in an incubator, the dried parts were crushed in a mixer grinder, and fine dust (800 gm) was collected for the experimentation. Dried and finely powdered dust (800 gm) was dissolved in hexane: isopropanol (3:2; v/v) on an air-tight glass jar and allowed for shaking in incubation at 37°C for 24 hours. After incubation, this solution was filtered through Whatman No.1 filter paper, and the filtrate was collected. Then the filtrate was evaporated by a rotary evaporator at 40°C. The collected part was content total oil ¹.

Chemicals: Major biochemical parameters were measured using diagnostic kits like urea, creatinine, glutamine oxalic transaminase (GOT), glutamate pyruvate transaminase (GPT), and alkaline phosphatase (ALP), C-reactive protein (CRP) purchased from Agappe Hills, Ernakulam, Kerala, India. We used Enzyme-Linked Immunosorbent Assay (ELISA), kits like interleukin-18 (IL-18), tissue injury molecule-1 (TIM-1), and interleukin- 10 (IL-10) purchased from Thermofisher Scientific, Invitrogen Bio Services India Pvt. Ltd. Bangalore, India. Paracetamol (neomol®) was purchased from a local market, Medinipur. All other chemicals like pyrogallol, tris buffer, trichloroacetic acid (TCA), thiobarbituric acid (TBA) and hydrochloric acid (HCL), sucrose, hydrogen peroxide (H2O2), ethanol, paraffin wax, hematoxylin, eosin, xylene, potassium dihydrogen phosphate (KH₂PO₄), sodium hydrogen phosphate (Na₂HPO₄) were purchased from SRL, India, and MERCK, India, sd FINE-CHEM Limited, India, HiMedia Laboratories Pvt. Ltd. Mumbai, India, and Crest Biosystems Goa, India.

Instruments: Biochemical parameters like urea, creatinine, GOT, GPT, ALP, CRP were measured by Semiautoanalyser (AGAPPE), antioxidant enzyme parameters like superoxide dismutase (SOD), catalase (CAT), and oxidative stress markers malondialdehyde (MDA) were measured by absorbance UV-VIS double beam Spectrophotometer (systronics, India), inflammatory markers were measured by ELISA plate reader (Thermofisher Scientific), TO was extracted using by Electrical Blender (Philips, India), soxhlet apparatus (Yoma, India), Microcentrifuge (Remi, India), BOD incubator, Incubator with shaker (Indian Instruments Ltd.). Microtome (MRM-RM, Safire, Coimbatore, Tamil Nadu), Inverted Biological Microscope (Magnus Opto Systems India Pvt. Ltd., Noida, Uttar Pradesh, India) for histopathological studies. All materials weighted by Digital weight balance (Accuracy-0.1mg) Adhair Dutta and Sons.

Selection of Animals and Care: Adult male albino outbred Wistar rats weighing 150 -170 g were obtained from the Centre for Translational Animal Research, Bose Institute, Kolkata. The animals were acclimatized to laboratory conditions for 2 weeks before experimentation. Animals were housed in three rats/cages in a temperature-controlled room (22± 20C) with 12–12 h dark-light cycles (8.00–20.00 h light, 20.00– 8.00 h dark) at a humidity of 50 ±10%.

All experimental procedures on animals follow the guidelines on the regulation of scientific experiments on animals stated by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), 2018 and our Institutional Animal Ethics Committee (IAEC) (13/IAEC (05)/RNLKWC/2019) ¹⁰.

Experimentally Induction of Oxidative Stress and Inflammation: Oxidative stress and inflammation (OS and I) were prompt through intraperitoneal injection of 0.2 ml freshly prepared paracetamol (N- acetyl- para- aminophenol, APAP) (15 mg/kg body weight) for 14 days. APAP (15 mg/kg b.w.) was prepared by mixing 2 ml 300 mg paracetamol (neomol®) with 2 ml normal saline water (v/v). At the induction time, vehicle control rats were injected with 0.2 ml carboxymethyl cellulose (CMC) alone.

Experimental Design: In this experiment, 48 rats were divided into 8 groups comprising 6 animals each. The TO was dissolved in 2 % CMC in distilled water and administered orally for 14 days.

Group I (normal control)- rats received a normal diet and water for 14 days.

Group II (vehicle control)- rats were given a normal diet and 0.2 ml of CMC for 14 days. Group III, IV, V, VI, VII, VIII (under OS and I)- rats were subjected to a normal diet and paracetamol at the dose of 15 mg/ kg body weight/ day for 14 days,

Group IV, V, VI, VII (treatment with TO)- rats orally received tea oil at the dose of 100, 200, 400, 800 mg/kg body weight/ day dissolved in CMC, respectively for 14 days,

Group VIII (treatment with olive oil)- orally administered of olive oil at the dose of 400 mg/kg body weight/ day dissolved in CMC for 14 days.

Animal Sacrificed and Plasma and Organ Collection: This experimental design was continued for 14 days. On the 15th day, the animals were sacrificed to collect blood samples from the aorta for haematological and biochemical analyses of renal plasma markers, hepatic functional markers, antioxidant enzyme markers, oxidative stress markers, and inflammatory cytokines. After blood collection, the specific organs like liver, kidney, and spleen were dissected, and weighed. Samples of organs were used for different biochemical analyses and some portions of organs were fixed in 10% formalin solution for 24 hours and then transferred into 70% alcohol for histopathological examination. The collected blood was centrifuged at 3000 r/min, and plasma fractions were separated for biochemical parameter assessment.

Estimation of Plasma Renal Markers:

Plasma Urea and Creatinine Level: The Plasma urea (mg/dl) and creatinine (mg/dl) levels were measured according to our earlier laboratory-established method by the semi-auto analyzer (AGAPPE, India) using standard AGAPPEE kit ^{11, 12}.

Estimation of Hepatic Functional Marker:

Biochemical Estimation of GOT, GPT and ALP Level in Plasma: For the assessment of hepatic function in plasma, we measured GOT, GPT, and ALP levels by our earlier established protocol by the semi-auto analyzer (AGAPPE, India) using kit manual protocol ^{11, 13}.

Estimation of Antioxidant Enzymes Activity and Lipid Peroxidation Levels:

Biochemical Assay of SOD Activity in Liver Tissue: The liver tissues were homogenized in ice-cold 100 mMTris-cacodylate buffer to give a tissue concentration of 50 mg/mL and centrifuged at 10,000 rpm for 20 mins at 4°C. The SOD activity of the supernatant was estimated by measuring the percentage of inhibition of the pyrogallol autooxidation by SOD according to our established method. One unit of SOD was defined as the enzyme activity that inhibited the autooxidation of pyrogallol by 50% ^{11, 12}.

Biochemical Assay of CAT Activity in Liver Tissue: CAT was measured biochemically in tissue supernatant by our laboratory-established method, and absorbance reading was noted using a spectrophotometer at 240 nm. For the evaluation of CAT in the liver tissues were homogenized separately in 0.05 M Tris Hydrochloric acid (HCl) buffer solution (pH-7.0) at a tissue concentration of 50 mg/mL. These homogenates were centrifuged separately at 10,000 rpm at 4°C for 10 minute ^{11, 12}.

Lipid Peroxidation Level: Lipid peroxidation in terms of malondialdehyde level was estimated according to our earlier studies using the thiobarbituric acid assay. The liver tissues were homogenized separately at a tissue concentration of 50 mg/mL in 0.1 M of ice-cold phosphate buffer (pH = 7.4). These homogenates were centrifuged at 10,000 rpm at 4°C for 5 min. Supernatants were used for the estimation of MDA. The MDA present within the sample was calculated using the extinction coefficient of 1.56 × 105 M/cm and expressed as the unit of nM/mg of tissue ¹⁴.

Estimation of Pro-inflammatory Markers:

Levels of IL-18: IL-18 (pg/dl) was measured by ELISA plate reader by our laboratory established method as per information provided in the kit manual (Thermofisher Scientific, Invitrogen Bioservices India Pvt. Ltd) ^{15, 16}.

Levels of TIM-1: TIM-1 (pg/dl) of plasma was measured by using a commercially available standard kit (Thermofisher Scientific, Invitrogen Bioservices India Pvt. Ltd) with standard protocol by ELISA (Thermofisher Scientific) plate reader in our laboratory ¹².

In brief, samples and standard were piped in 96 well plates that were pre-coated with specific antibodies for TIM-1. In the next step, wells were washed, and a biotinylated antibody specific for TIM-1 was added and incubated for 1 hour. After that, the wells were washed, and streptavidin- HBP solution was added and followed by washing, and the solution was read at 450 nm using an ELISA plate reader.

Levels of CRP: CRP (mg/L) level of plasma was measured by a semi-auto analyzer (AGAPPE, India) using the information provided in the kit manual (AGAPPE, India) ¹⁷.

Estimation of Anti-inflammatory Markers:

Levels of IL-10: The quantitative determination of cytokines like IL-10 (pg/dl) of plasma was performed by a commercially available standard kit (Thermofisher Scientific, Invitrogen Bioservices India Pvt. Ltd) with standard protocol by ELISA (Thermofisher Scientific) plate reader in our laboratory. Samples and standards were pipetted into the wells that been have pre-coated with specific antibodies for IL- 10. In the next step, the wells were washed and enzyme-linked antibodies specific for IL-10 were added to wells and incubated. Then, the wells were aspirated and washed. The process was repeated three times. After the last washing, aspiration was performed to remove any remaining wash buffer. The plate was inverted against clean paper towels and the solution was read at 450 nm using a microplate reader ².

Histopathological Studies: Liver tissues were embedded in paraffin wax and 5 µm sections were prepared with a rotary microtome. These thin sections were stained with hematoxylin and eosin (H and E), mounted on glass slides, and observed for pathological changes using an Inverted microscope ¹⁶.

Statistical Analysis: Data were expressed as mean ± SE (n=6). One-way analysis of variance (ANOVA) followed by multiple two-tail t-tests to detect inter-group differences and bars with different superscripts (*, **) differ from each other significantly (p< 0.05)¹⁸.

RESULTS:

Bodyweight (b.w.) and Somatic Index of Liver, Kidney, and Spleen: The present study reveals that body weight significantly (p < 0.05) increased at the end of the experiment in normal control, vehicle control, OS, and I rats treated with TO at different doses of 100, 200, 400 and 800 mg/kg of b. w./day dissolved in CMC as well as when treated with olive oil at the dose of 400 mg/kg b.w./ day in comparison with only OS and I group. The introduction of high-dose APAP decreases the body weight compared to a normal control group. We have important findings that there are no significant (p > 0.05) changes in the weight of organs like the liver, kidney, and spleen in all groups of rats Table 1.

TABLE 1: EFFECTS OF TEA OIL ON BODY WEIGHT AND RELATIVE ORGAN WEIGHT ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTAR RATS

Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-test. *significant difference (p < 0.05) vs control (Group I); ** significant difference (p < 0.05) vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400, and 800 mg/kg of b. w./day dissolved in CMC respectively, Group VIII- APAP-induced OS and I + co-administration of olive oil at 400 mg/kg of b. w./ day. ↑- % of body weight increased, ↓- % of body weight decreased

Effects of Tea Oil on the Levels of Urea in Plasma: The level of urea in plasma is found to increase significantly (p < 0.05) in high dose APAP induced OS and I rats than in the normal controls rats. However, with the addition of TO and olive oil in high dose APAP induced OS and I, the level of urea in plasma significantly (p < 0.05) decreased in comparison with only APAP- induced OS and I rats. Surprisingly the addition of TO (at the dose of 400 mg/kg b.w./ day dissolved in CMC) not only decreases the level of urea in plasma but also reverts to the normal control value at a certain dose Fig. 1.

FIG. 1: EFFECTS OF TEA OIL ON UREA LEVEL IN PLASMAON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05)vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400 and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

Effects of Tea Oil on the Levels of Creatinine in Plasma: The same observations were found when the level of creatinine in plasma was determined that the level of creatinine significantly (p < 0.05) increased in the case of APAP-induced OS and I rats than the normal control rats.

But the administration of TO (at the dose of 100, 200, 400, and 800 mg/kg of b. w./day dissolved in CMC, respectively), significantly (p < 0.05) decreased the levels of creatinine in plasma than the only APAP- induced OS and I rats Fig. 2.

FIG. 2: EFFECTS OF TEA OIL ON CREATININE LEVEL IN PLASMA ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05) vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400, and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at the dose of 400 mg/kg of b.w./ day.

Effects of Tea Oil on the Levels of GOT, GPT in Plasma: The activities of plasma GOT, GPT levels were significantly (p < 0.05) higher in only APAP-induced OS and I rats than the normal control rats. But the addition of TO (at the dose of 100, 200, and 800 mg/kg of b. w./ day dissolved in CMC, respectively) and olive oil (at the dose of 400 mg/kg of b.w./ day) helped to decrease the level of plasma GOT and GPT than the only OS and I rats. Results show that the threshold value achieved for the addition of TO (at the dose of 400 mg/kg b.w./ day dissolved in CMC) decreases the GOT and GPT levels, reaching the value of normal control Fig. 3, 4.

FIG. 3: EFFECTS OF TEA OIL ON GOT LEVEL IN PLASMAON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-test.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05) vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400 and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

FIG. 4: EFFECTS OF TEA OIL ON GPT LEVEL IN PLASMAON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05)vs OS and I (Group III).

Effects of Tea Oil on the Levels of ALP in Plasma: The activities of plasma ALP levels were significantly (p < 0.05) reduced in addition to TO (at the dose of 100, 200, 400, and 800 mg/kg of b. w./ day dissolved in CMC respectively) and olive oil (at the dose of 400 mg/kg of b.w./ day) treated rats in comparison with only OS and I rats. ALP plasma level was significantly (p < 0.05) higher in OS and I rats than the normal control rats Fig. 5.

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400 and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

FIG. 5: EFFECTS OF TEA OIL ON ALP LEVEL IN PLASMAON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-test.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05) vs OS and I (Group III).

Effects of Tea Oil on the activities of SOD Level in Liver Tissue: Administration of TO (at the dose of 100, 200, 400 and 800 mg/kg of b.w./ day dissolved in CMC, respectively) on APAP- induced OS and I rats and administration of olive oil (at the dose of 400 mg/kg of b.w./day), normal and vehicle control rats, we observed significant (p < 0.05) elevation of SOD level in liver tissue than the APAP-induced OS and I rats. The efficacy of this enzyme was significantly (p < 0.05) decreased in liver tissue in APAP-induced OS and I rats in comparison to normal control rats. APAP- induced OS and I with co-administration of TO (at the dose of 100, 200, 400, and 800 mg/kg of b.w./ day dissolved in CMC, respectively) to the rat’s significant alteration of SOD level than the APAP- induced OS and I rats and the level of SOD activity was reverting to normal control value by the addition of TO (at the dose of 400 mg/kg of b.w./ day dissolve in CMC) Fig. 6.

FIG. 6: EFFECTS OF TEA OIL ON SOD IN LIVER TISSUE ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05)vs control (Group I); **Significant difference (p <0.05)vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400, and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

Effects of Tea Oil on the activities of CAT Level in Liver Tissue: Similar findings we observed that TO (at the dose of 100, 200, 400, and 800 mg/kg of b.w./ day dissolved in CMC, respectively) treated on APAP- induced OS and I rat the activities of CAT in liver tissue were significantly (p < 0.05) increased in comparison with only APAP-induced OS and I rats.

In APAP-induced OS and I rat, the activities of this enzyme were significantly (p < 0.05) reduced than the normal control rats. There was a significant alteration of CAT level when the administration of TO (at the dose of 100, 200, 400, and 800 mg/kg of b.w./ day dissolved in CMC, respectively) in rats, in comparison to APAP- induced OS and I rat Fig. 7.

FIG. 7: EFFECTS OF TEA OIL ON CAT IN LIVER TISSUE ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05)vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400, and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

Effects of Tea Oil on Lipid Peroxidation Level: The lipid peroxidation level was significantly (p < 0.05) higher in liver tissue in APAP-induced OS and I rats in comparison with normal and vehicle control rats, APAP- induced OS and I treated with TO, and olive oil rats. But in APAP- induced OS and I rats that received TO at various doses, the MDA level was significantly (p < 0.05) decreased in the liver, and the MDA level was seen to be similar to the TO (at the dose of 400 mg/kg b.w./ day dissolve in CMC) treated rats in comparison with normal control rats Fig. 8.

FIG. 8: EFFECTS OF TEA OIL ON MDA LEVEL IN LIVER TISSUE ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05)vs control (Group I); **Significant difference (p <0.05)vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400, and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

Effects of Tea Oil on IL-18 Level in Plasma: The present study showed that the pro-inflammatory cytokines like IL-18 were significantly (p < 0.05) increment in plasma in only APAP-induced OS and I rats compared with normal as well as vehicle control rats. Administration of TO on APAP- induced OS and I group and administration of olive oil, this IL-18 level was gradually decreased in plasma in comparison with only APAP- induced OS and I rats and APAP- induced OS and I TO (at the dose of 400 mg/kg b.w./ day dissolve in CMC), IL-18 level was similar to the normal control rats Fig. 9.

FIG. 9: EFFECTS OF TEA OIL ON IL-18 LEVEL IN PLASMA ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05) vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400, and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

Effects of Tea Oil on TIM-1 Level in Plasma: The levels of TIM-1 were significantly (p < 0.05) higher in plasma in APAP- induced OS and I rats compared with normal control rats. Administration of TO at different doses on APAP-induced OS and I rats and also the administration of olive oil this TIM-1 level was gradually reduced in plasma to compare with APAP-induced OS and I group, and this TIM-1 level was found to be similar to TO (at the dose of 400 mg/kg b.w./ day dissolve in CMC) treated rats than the normal control rats Fig. 10.

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400 and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at 400 mg/kg of b.w./ day.

FIG. 10: EFFECTS OF TEA OIL ON TIM-1 LEVEL IN PLASMA ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-test.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05) vs OS and I (Group III).

Effects of Tea Oil on CRP Level in Plasma: The CRP level in APAP- induced OS and I rats was significantly (p < 0.05) increased to compared with normal control and other TO-treated APAP-induced OS and I groups. Administration of TO on APAP- induced OS and I rats and administration of olive oil, this CRP level was gradually decreased in plasma to compared with only APAP- induced OS and I rat Fig. 11.

FIG. 11: EFFECTS OF TEA OIL ON CRP LEVEL IN PLASMA ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05)vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400 and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at the dose of 400 mg/kg of b.w./ day.

Effects of Tea Oil on Anti-inflammatory Cytokines (IL-10) Level in Plasma: The present study were showed that the anti-inflammatory cytokines like IL-10 was significantly (p < 0.05) decreased in plasma in APAP- induced OS and I rats when compared with group I normal control rats.

Administration of TO at different doses on APAP- induced OS and I group and also the administration of olive oil this IL-10 level was gradually increased in plasma to compare with APAP- induced OS and I rats and this IL-10 level was seen resettled to the TO (at the dose of 400 mg/kg b.w./ day dissolve in CMC) treated rats to compared with normal control rats Fig. 12.

FIG. 12: EFFECTS OF TEA OIL ON IL-10 LEVEL IN PLASMA ON PARACETAMOL-INDUCED OS AND I IN MALE ALBINO OUTBRED WISTER RATS. Data are expressed as Mean ± SE (n=6 rats in each group). Statistical significance was evaluated by one-way analysis of variance (ANOVA) followed by multiple two-tail t-tests.* Significant difference (p <0.05) vs control (Group I); **Significant difference (p <0.05) vs OS and I (Group III).

Group I- Normal control, Group II- Vehicle control, Group III- Paracetamol- induced OS and I, Group IV, V, VI, VII- Paracetamol- induced OS and I + co-administration of TO at the dose of 100, 200, 400 and 800 mg/kg of b. w./ day dissolve in CMC respectively, Group VIII- APAP- induced OS and I + co-administration of olive oil at the dose of 400 mg/kg of b.w./ day.

Effects of Tea Oil on the Hepatohistopathology of Liver Tissue: From Fig. 13 we observed the histopathological architecture of liver tissue.

From Fig. 7, section A (Control) showed the normal architecture of the central vein vertically with diameters 3.00 μm and adjoining hepatic cells.

The paracetamol administration group rats (Section C, APAP-induced OS and I) showed severe degeneration of hepatic cells surrounding the central vein. Also, they damaged the architecture of the central vein vertically with a diameter 6.03μm.

The liver tissue of rats which received paracetamol with co-administration of TO at the dose of 100 mg (Section D), 200 mg (Section E), and 800 mg (Section G)/kg body weight/day showed mild changes in hepatic cells and the diameter of the central vein vertically with diameter 4.53 μm, 4.12 μm and 4.19μm respectively.

But the liver tissue of rats that received paracetamol with TO at the dose of 400 mg/ kg body weight showed the density of the central vein vertically with a diameter 2.90 μm and normal orientation of hepatic cell (Section F).

In contrast the liver tissue of rats that received paracetamol with olive oil, there was no change of central vein vertically with a diameter 5.93 μm as well as hepatic cells (Section H) in comparison with paracetamol treated rats and also TO treated rats.

FIG. 13: HISTOPATHOLOGICAL ANALYSIS OF LIVER TISSUE MEASURED VERTICALLY USING AN INVERTED MICROSCOPE

DISCUSSION: Paracetamol overdose creates high oxidative stress in the liver by decreasing SOD and CAT to induce hepato-toxicity. Overdose of paracetamol (N- acetyl- para- aminophenol, APAP) (15 mg/kg body weight) is generally given to induce oxidative stress and inflammation (OS and I) in albino outbred Wistar rats ¹³. In this study, we are mainly concentrating to evaluate the effects of tea oil (TO) on a high dose of APAP-induced OS and I in a rat model by observing the changes that occurred in the activities of antioxidant enzyme activity, oxidative stress marker, expression of pro and anti-inflammatory markers and hepatohistopathology study.

The diminution of antioxidant levels and increment in metabolite concentration by induction of high dose APAP causes some physiological abnormalities like loss of body weight, adduction of cytotoxic protein, and necrosis of hepatic and renal cells ^{19, 20}. Remarkable loss in body weight was found in OS and I rats, probably due to muscle protein destruction. But surprisingly we observe that oral administration of TO in different doses not only changes the weight loss but also improves body weight appreciably.

The distinctive parameters of hepatotoxicity and renotoxicity markers are generally determined from the urea and creatinine levels in plasma ²¹. Overdose of APAP- induced OS and I also so the increased level of urea and creatinine in plasma which were considered the remarkable biomarkers of renal malfunction or renal disruption ²². In this present study, we find the plasma urea and creatinine level were significantly (p < 0.05) increased while intraperitoneally administered the high dose APAP-induced OS and I rats due to the application of high dose APAP have been significantly decreased the plasma urea and creatinine level when the introduction of TO at different doses respectively. APAP-induced renal damage is known to causes the degeneration of epithelial cell lines and dilation of glomerular kidney tissue but the oral administration of TO help to reduce the plasma urea and creatinine level through modulating inflammatory cytokines which is the possible mechanism of nephroprotective activity ²³. Similar outcomes were found by other researchers who observed that plant oil from ginger and turmeric rhizome decreased the level of renal biomarkers like urea, and creatinine.

Normally GOT, GPT, and ALP enzymes are present in low concentrations in plasma. When these enzymes were emancipating the bloodstream and elevated the level which indicates that the hepatopathy and the endothelial cell layer were damaged ²⁴. Evaluation of liver function enzymes can be done by the analysis of GOT that is present in mitochondria, GPT, and ALP, which is present in the cytoplasm of a liver cell at higher concentrations ²⁵.

Administration of APAP in overdose results in oxidative damage, necrosis of hepatic cells, hepatic glutathione depletion and infiltration of inflammatory cytokines, hepatocyte vacuolization, and mitochondrial dysfunction, promoting the elevation in plasma level of enzymes transferases such as GOT, GPT, and ALP ⁶. In the present study, it is observed that high dose APAP increases the value of GOT, GPT, and ALP in the plasma of the rat, but the group of rats which received APAP along with TO in different doses showed a significant (p < 0.05) reduction of those enzymes level in plasma by controlling the damage of hepatocellular tissues. According to Grespan et al., Thymus vulgaris plant oil significantly decreased GOT, GPT, and ALP levels in plasma compared with APAP-induced injury rats ²⁶. Plant oil protects the functional integrity of the plasma membrane as well as increment the regenerative capacity of the liver ²⁷. So, it can be said that TO protect the liver from APAP-induced liver damage. At the same time, SOD and CAT are known to be the most important free radical scavenging enzymes involved in enhancing the effects of oxygen metabolism and are the main protective enzymes that counter oxidative stress ^{28, 29}.

Results also show that SOD and CAT in liver tissue significantly (P < 0.05) decreased significantly in OS and I, compared to normal control rats. The depletion of the activities of SOD and CAT indicates periodic production of free radicals in profusion amount that results in cellular damage. However, in TO treated group, there was a significant increase (P < 0.05) in SOD and CAT levels. TO, riched in linolenic acid, biologically counteract. CAT is the most important antioxidant that protects the cell from lipid hydroperoxide formation ³⁰. From various studies, we observed that administration of plant oils of clove, cumin, and fennel on rat models, increases the level of SOD, CAT compared to normal control rats ³¹.

When the generation of free radicals exceeds the body's antioxidant level, lipid peroxidation occurs, which can be estimated from the level of MDA, which is a reliable sign of the degree of lipid peroxidation. Polyunsaturated fatty acids such as linolenic acid act as a protective constituent against lipid peroxidation by increasing the cellular anti-oxidants level ³². MDA level was significantly (P < 0.05) increased in OS, and I rats, while this level gradually decreased when the rats received TO. From similar studies, we observed that administration of lavender oil and other plant oil at various concentrations decreases the lipid peroxidation level and also acts positively against various diseases that occur from selecting oxidative stress ². During inflammation, monocytes turn into chemokines, initiate the lesion, and are converted into macrophages that undergo phagocytosis and release various pro-inflammatory cytokines like IL-18 and TIM-1. Pro-inflammatory cytokines like IL-18, TIM-1 and anti-inflammatory cytokines such as IL-10 play a vital role in the inflammatory process. From another study, we observed that plant oil reduces the stage of respiratory inflammation by preventing the production of IL-18, TIM-1 in epithelial cell lining. Plant oil acts as an anti-inflammatory agent by suppressing the inflammatory enzymes and secretion of pro-inflammatory cytokines like IL-18, TIM-1 or down-regulating the expression of inflammatory mediators. The activities of inflammatory enzymes are inhibited by NSAIDs which lead to the induction of inflammatory responses ³³.

Our result showed that in APAP- induced OS and I rats, pro-inflammatory cytokine levels were significantly (P < 0.05) when compared with normal control and other TO-treated groups. In contrast, after receiving TO at different doses, pro-inflammatory cytokines such as IL-18 and TIM-1 level was significantly (P < 0.05) decreased in plasma compared to only APAP-induced OS and I rats. Chen et al., observed that plant oil from Cinnamomum camphora (Linn.) markedly decreased the inflammatory cytokines, including IL-18 and TIM-1 levels ^{34, 35}. IL-10 is commonly known as anti-inflammatory and immunosuppressive cytokines, which upregulates the anti-inflammatory cytokines and inhibits the functions of pro-inflammatory cytokines like IL-18, which has been established in various studies ³⁶.

In the present study, TO significantly (P < 0.05) increased the level of anti-inflammatory cytokines like IL-10. Various studies indicated that plant oil from Trachydium roylei, lavender oil, and tea tree oil enhances anti-inflammatory cytokine IL-10 levels in dose depending on a mechanism ^{2, 4, 7}. The liver is the vital organ that plays an important role in metabolism, excretion, and detoxification ³⁷. APAP causes liver cirrhosis and liver damage by increasing lipid peroxidation in the liver resulting in inflammation. In inflammation, Kupffer cells form different cytokines, which take a crucial role in causing the degeneration of hepatocytes. But TO, rich in polyunsaturated fatty acids and various antioxidants, improve the hepatic cell structure by improving liver function. According to EI-Hadary et al., clove oil decreases hepatocyte necrosis, hepatic lipidosis, and central lobular necrosis ³⁸. After a thorough investigation, the study shows that the effective dose of TO was 400 mg/kg body weight/ day dissolved in CMC out of three doses used for the study. From the various results, we observed that tea oil acts effectively in this dose. Although the present study focuses for the first time on the anti-oxidative and anti-inflammatory effects of Cammelia sinensis L. oil against APAP- induced OS and I in a rat model, there were some limitations of the present work. The investigation is needed to ascertain some inflammatory markers such as cycloxygenase (COX), lipooxygenase (LOX), prostaglandins, leukotrienes for the possible mechanism of action in the inflammation and also needed some other constituents in Cammelia sinensis L. oil except fatty acid profile (already identified in previous work). In our next study, we will try to identify and overcome the limitation of the present work. From the present work, we suggest that Cammelia sinensis L. oil is beneficial in improving oxidative stress-activated chronic inflammatory diseases.

CONCLUSION: It can be concluded from the findings mentioned in the present study that tea oil shows potent antioxidative activities against high-dose paracetamol-induced oxidative stress and inflammation by regulating the levels of pro-inflammatory and anti-inflammatory cytokines. So it is established that tea oil acts as an effective agent for preventing different inflammatory diseases.

Ethics Approval and Consent to Participate: All experimental procedures on animals were in accordance with the guidelines on the regulation of scientific experiments on animals stated by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), 2018 and our Institutional Animal Ethics Committee (IAEC)(13/IAEC (05)/RNLKWC/2019), Raja N.L. Khan Women’s College (Autonomous), Midnapore.

Authors’ Contributions: Meghamala Mandal has made a contribution to the analysis and prepared the original manuscript. Shrabanti Pyne, Supriya Bhowmik and Deblina Giri performed the laboratory experiments. Dr. Koushik Das contribute to designing the study and reviewing the article content critically, and Dr. Jayasree Laha reviewed the article content critically. All authors have read manually and approved the final manuscript.

ACKNOWLEDGEMENT: Not applicable

CONFLICTS OF INTEREST: The authors declare that there are no conflicts of interest

REFERENCES:

1. Mandal M, Das K, Nandi DK, Das B and Sreenivas RJ: Analysis of fatty acids from oil of green tea (Camellia sinensis) by Gas Chromatography coupled with flame ionization detector and its anticancer and antibacterial activity In vitro. International Journal of Pharmacy and Pharmaceutical Sciences 2019; 11(3): 104-11.https://doi.org/10.22159/ijpps.2019v11i3.29888
2. Aboutaleb N, Jamali H, Abolhasani M and Toroudi HP: Lavender oil (Lavandula angustifolia) attenuates renal ischemia/reperfusion injury in rats through suppression of inflammation, oxidative stress and apoptosis. Biomedicine and Pharmacotherapy 2019; 110: 9-19. 10.1016/j.biopha.2018.11.045
3. Marchio P, Guerra-Ojeda S, M. Vila J, Aldasoro M, Victor MV and Mauricio MD: Targeting Early Atherosclerosis: A Focus on Oxidative Stress and Inflammation. Oxidative Medicine and Cellular Longevity 2019; 2019: 1-32. https://doi.org/10.1155/2019/8563845
4. Wang YT, Zhu L, Zeng D, Long W and Zhu SM: Chemical composition and anti-inflammatory activities of essential oil from Trachydiumroylei. Journal of Food and Drug Analysis 2016; 24: 602-9. http://dx.doi.org/10.1016/j.jfda.2016.02.009
5. Cervantes-Gracia K, Raja K, Llanas-Cornejo D, N. Cobley J, L. Megson I, Chahwan R and Husi H: Oxidative stress and inflammation in the development of cardiovascular disease and contrast induced nephropathy. Vessel Plus 2020; 4(27): 1-19. 10.20517/2574-1209.2020.22
6. Cardia G, Silva-Comar S, Silva E, Rocha E, Comar J, Silva-Fiho S, Zagotto M, Uchida NS, Bersani-Amado CA, and Cuman RKN: Lavender (Lavandula officinalis) essential oil prevents acetaminophen-induced hepatotoxicity by decreasing oxidative stress and inflammatory response. Research Society and Development 2021; 10(3): e43410313461. http://dx.doi.org/10.33448/rsd-v10i3.13461
7. Baldissera MD, Souza CF, Doleski PH, Vargas AguedaCde, Duarte MMMF, Duarte T, Boligon AA, Leal DBR and Baldiserotto B: Melaleucaalternifolia essential oil prevents alterations to purinergic enzymes and ameliorates the innate immune response in silver catfish infected with Aeromonashydrophila Microbial Pathogenesis 2017; 109: 61-66. http://dx.doi.org/10.1016/j.micpath.2017.05.026
8. A BS, Kang JH, Yang H, Jung EM, Kang HS, Choi IG, Park MJ and Jeung EB: Anti-inflammatory effects of essential oil from Chamaecyparis obtuse via the vyclooxygenase-2 pathway in rats. Molecular Medicine Reports 2013; 255-59. doi.org/10.3892/mmr.2013.1459
9. Das B, Biswas B and Ghosh D: Duration dependent antiapoptotic efficacy of Camellia sinensis (green tea) on streptozotocin induced diabetes linked testicular hypofunction in albino rat: genomic and flow cytometric assessment. International Journal of Pharmacy and Pharmaceutical Sciences 2019; 11(6): 85-93. http://dx.doi.org/10.22159/ijpps.2019v11i6.31150
10. Committee for the purpose of Control and Supervision of Experiments of Animals (CPCSEA): Compendium of CPCSEA, Ministry of Environment, Forest and Climate Change. Government of India 2018.
11. Das K, Roy S, Mandal S, Pradhan S, Debnath S, Ghosh D, and Nandi DK: Renal protection by isolated phytocompounds from Termina liaarjuna bark fraction on dehydration induced uremia, oxidative stress and kidney disease rats. International Journal of Recent Scientific Research 2016; 7(10): 13787-95. http://recentscientific.com/sites/default/files/Article%20-6333pdf
12. Giri D, Nandi DK, and Das K: Establishment of novel urinary kidney disease new biomarkers and therapeutic effect of methanol fraction of Termina liaarjuna on acetaminophen induced kidney disease in rats. Asiaan Journal of Pharmaceutical and Clinical Research 2019; 12(3): 117-24. https://doi.org/10.22159/ajpcr.2019.v12i3.28485
13. Roy S, Pradhan S, Das K, Samanta A, Mandal A, Mandal S, Patra A, Samanta A, Sinha B and Nandi DK: Acetaminophen Induced Kidney Failure in Rats: A dose response study. Journal of Biological Sciences 2015; 15(4): 187-93. 10.3923/jbs.2015.187.193
14. Pradhan S, Mandal S, Roy S, Mandal A, Das K and Nandi DK: Attenuation of uremia by orally feeding alpha-lipoic acid on acetaminophen induced uremic rats. Saudi Pharmaceutical Journal 2013; 21: 187-92. 10.1016/j.jsps.2012.03.003
15. Zhang C, Bi Y, Jin G, Gan H and Yu L: High and fluctuating glucose levels increase the expression and secretion of interleukin 18 in mouse peritoneal macrophages. Molecular Medicine Reports 2015; 12: 2715-220. 10.3892/mmr.2015.3753
16. Khatun H, Das K, Nandi DK and Chattopadhyay A: Supplementation of seed dust of Viciafaba and sesame ameliorates high lipid diet-induced dyslipidemia in rats. Current Research in Nutrition and Food Science 2019; 7(1): 1-17. http://dx.doi.org/10.12944/CRNFSJ.7.1.20
17. Belkhodja H, Meddah B and Gezici S: Anti-Inflammatory Effects of Essential Oils from Rosmarinusofficinalis and Populusalbaon Experimental Models of Acute and Chronic Inflammation in Rats. Indian Journal of Pharmaceutical Education and Research 2017; 51(3): 180-184. 10.5530/ijper.51.3s.8
18. Sokal RR and Rohle FJ: Introduction to analysis of variance. In: SokalRr, Rohle FJ edition. WH Freeman and Company, Biometry, New York 1997; 179-206.
19. Karigidi KO and Olaiya CO: Antidiabetic activity of corn steep liquor extract of Curculigopilosa and its solvent fractions in streptozotocin-induced diabetic rats. Journal of Traditional and Complementary Medicine 2019; 10 (2020): 555-64. https://doi.org/10.1016/j.jtcme.2019.06.005
20. Hussain Z, Khan JA, Arshad A, Asif P, Rashid H, and Arshad MI: Protective effects of Cinnamomum zeylanicum (Darchini) in acetaminophen-induced oxidative stress, hepatotoxicity and nephrotoxicity in mouse model. Biomedicine and Pharmacotherapy 2019; 109(2019): 2285-92. https://doi.org/10.1016/j.biopha.2018.11.123
21. Khalid S, Afzal N, Khan JA, Hussain Z, Qureshi AS, Anwar H and Jamil Y: Antioxidant resveratrol protects against copper oxide nanoparticle toxicity in vivo. Naunyn Schmiedeberg’s Archives of Pharmacology 2018; 391(10):1053-62. https://doi.org/10.1007/ s00210-018-1526-0
22. Nain P, Saini V, Sharma S and Nain J: Antidiabetic and Antioxidant Potential of Emblica officinalis Leaves extract in streptozotocin-induced type-2 diabetes mellitus (T2DM) rats. Journal of Ethnopharmacology 2012; 142 (1): 65-71. 10.1016/j.jep.2012.04.014
23. Javedan G, Shidfar F, Davoodi SH, Ajami M, Gorjipour F, and Sureda A: Conjugated linoleic acid rat pretreatment reduces renal damage in ischemia/reperfusion injury: Unraveling antiapoptotic mechanisms and regulation of phosphorylated mammalian target of rapamycin. Molecular Nutrition and Food Research 2016; 60(12): 2665–77. 10.1002/mnfr.201600112
24. Mandal A, Patra A, Mandal S, Roy S, Mahapatra TD, Mahapatra SD, Das K, Mondal KC and Nandi DK: Therapeutic potential of different commercially available synbiotic on acetaminophen-induced uremic rats. Clinical and Experimental Nephrology 2015; 19(2): 168-77. 10.1007/s10157-014-0971-4
25. Muhammad A, Aisha A, Azeem MA, Navia-ul-Zafar and Ahmad SI: Hepatoprotective effect of barrisal (herbal drug) on carbon tetrachloride induced hepatic damage in rats. African Journal of Pharmacy and Pharmacology 2013; 7(15): 776-84.
26. Grespan R, Aguiar RP, Giubilei FN, Fuso RR, Damião MJ, Silva EL, Mikcha JG, Harnandes L, Amado CB and Cuman RKN: Hepatoprotective Effect of Pretreatment with Thymus vulgaris Essential Oil in Experimental Model of Acetaminophen-Induced Injury. Evidence-Based Complementary and Alternative Medicine 2014; 1-8. http://dx.doi.org/10.1155/2014/954136
27. Pradeep K, Victor Raj Mohan C, Gobianand K and Karthikeyan S: protective effect of Cassia fistula Linn on diethylnitrosamine induced hepaticellular damage and oxidative stress in ethanol pretreated rats. Biological Research 2010; 43: 113-25.
28. Bellassoued K, Hsouna AB, Athmouni K, Pelt JV, Ayadi FM, Rebai T and Elfeki A: Protective effects of Mentha piperita L. leaf essential oil against CCl4 induced hepatic oxidative damage and renal failure in rats. Lipids in Health and Disease 2018; 17(1):1-14. DOI 10.1186/s12944-017-0645-9
29. Somani SM, Husain K, Whitworth C, Trammell GL, Malafa M, and Rybak L: Dose-dependent protection by lipoicacid against cisplatin-induced nephrotoxicity in rats: antioxidant defense system. Pharmacology and Toxicology 2000; 86(5): 234–41. 10.1034/j.1600-0773.2000.d01-41.x
30. Smith AR, Shenvi SV, Widlansky M, Suh JH and Hagen TM: Lipoic acid is a potential therapy for chronic diseases associated with oxidative stress. Current Medicinal Chemistry 2004; 11: 1135–46.
31. Sheweita SA, El-Hosseiny LS and Nashashibi MA: Protective Effects of Essential Oils as Natural Antioxidants against Hepatotoxicity Induced by Cyclophosphamide in Mice. Plos One 2016; 1-17. DOI:10.1371/journal.pone.0165667
32. Nagao T, Komine Y, Soga S, Meguro S, Hase T, Tanaka Y and Tokimitsu I: Ingestion of a tea rich in catechins leads to a reduction in body fat and malondialdehyde modified LDL in men. The American Journal of Clinical Nutrition 2005; 81(1): 122-9. 10.1093/ajcn/81.1.122
33. Gurpinar E, Grizzle WE and Piazza GA: NSAIDs inhibit tumorigenesis, but how? Clinical Cancer Research 2014; 20: 1104-13. 10.1158/1078-0432.CCR-13-1573
34. Chen J, Tang C, Zhou Y, Zhang R, Ye S, Zhao Z, Lin L, and Yang D: Anti-Inflammatory Property of the Essential Oil from Cinnamomum camphora (Linn.) Presl Leaves and the Evaluation of Its Underlying Mechanism by Using Metabolomics Analysis. Molecules 2020; 25(20): 4796-809. doi:10.3390/molecules25204796
35. Zhang Y, Li Q, Fang M, Ma Y, Liu N, Yan X, Zhou J and Li F: The Kidney Injury Induced by Short-Term PM2.5 Exposure and the Prophylactic Treatment of Essential Oils in BALB/c Mice. Oxidative Medicine and Cellular Longevity 2018; 2018; 1-12.
36. Chung F: Anti-inflammatory cytokines in asthma and allergy: interleukin-10, interleukin-12, interferon γ. Mediators of Inflammation 2001; 10(2): 51-9.
37. Roy S, Das K, Mandal S, Pradhan S, Patra A, Samanta A, Mandal A and Kar S and Nandi DK: Asparagus Racemosus Roots Ameliorates Acetaminophen Induced Hepatotoxicity in Rats: An Experimental, Biochemical and Histological Study. International Journal of Recent Scientific Research 2014; 5(6): 1192-7. http://dx.doi.org/10.13040/IJPSR.0975-8232.4(8).3004-12
38. El-Hadary A, and Ramadan Hassanien M: Hepatoprotective effect of cold-pressed Syzygium aromaticumoil against carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. Pharmaceutical Biology 2015; 54(8): 1364-72.
    

    ---

    How to cite this article:

    Mandal M, Pyne S, Bhowmik S, Giri D, Das K and Laha J: To study the antioxidant and anti-inflammatory properties of oil from leaves of Camellia sinensis L. in experimental animal models. Int J Pharm Sci & Res 2023; 14(1): 381-97. doi: 10.13040/IJPSR.0975-8232.14(1).381-97.

    ---

    All © 2023 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.