MICHELIA CHAMPACA LEAF EXTRACTS EXHIBIT HYPOGLYCEMIC EFFECT AND HYPOLIPIDEMIC ACTIVITY IN STREPTOZOTOCIN-NICOTINAMIDE-INDUCED DIABETIC RATS
HTML Full TextMICHELIA CHAMPACA LEAF EXTRACTS EXHIBIT HYPOGLYCEMIC EFFECT AND HYPOLIPIDEMIC ACTIVITY IN STREPTOZOTOCIN-NICOTINAMIDE-INDUCED DIABETIC RATS
Jyoti Nanda *, Munesh Mani , Shanti Bhushan Mishra and Neeraj Verma
School of Pharmaceutical Sciences, CSJM University, Kanpur, Uttar Pradesh, India.
ABSTRACT: A metabolic disease called diabetes mellitus in which blood glucose levels are abnormally high. It could be due to tissue glucose intolerance and impaired insulin production. In a Type II diabetes model generated by Streptozotocin and nicotinamide, the antidiabetic potential of ethanolic and aqueous leaf extracts of Michelia champaca (Magnoliaceae) was compared to a control group of diabetes. The anti-diabetic activity was tested on Wistar rats. The effective antihyperglycemic ethanolic and aqueous leaf extracts were tested at two doses of 250 mgkg-1 b.w. and 500 mgkg-1 b.w. for 21 days. Compared to the diabetic control group, the ethanolic extract at a dose of 500 mgkg-1 b.w. exhibits a highly significant (p<0.001) reduction in fasting blood glucose levels. After the trial, the blood sample was drawn from all animals for biochemical analysis. In extract-treated diabetic rats, the serum lipid profile was calculated. The biochemical investigation, serum cholesterol, serum triglyceride, high-density lipoprotein, and low-density lipoprotein support the ethanolic extract's anti-diabetic effectiveness.
Keywords: Michelia champaca, Biochemical, Streptozotocin-Nicotinamide, Insulin, Antidiabetic activity
INTRODUCTION: Hyperglycemia is a collection of illnesses marked by high blood sugar level, altered lipid, carbohydrate and protein metabolism, and an elevated risk of vascular consequences. Type II diabetes affects the vast majority of people. According to the WHO Global Report on Diabetes 2016, people with hyperglycemia have nearly doubled to 422 million 1. There are diabetic patients that have insulin insufficiency or insulin resistance, as well as excessive glucagonemia.
There is an increase in hepatic glucose synthesis, a decrease in peripheral glucose uptake and a reduction in glucose to glycogen conversion in the liver 2. In Type 2 diabetes, beta-cells are remaining no longer to produce a sufficient amount of insulin. It arises due to sedentary lifestyle, alcohol consumption, cigarette smoking, physical inactivity and genetics which cause overweight and obesity leads to a reduction in insulin sensitization and glucose tolerance 3.
Patients with diabetic kidney disease (DKD) are at increased risk of end-stage kidney disease (ESKD) and cardiovascular disease, increased mortality, lower quality of life and higher health care costs 4. Reactive oxygen species induce a complexing agent called advanced glycation end-product (AGE), which may convert normal glucose level stage to hyper glucose level stage 5. Flavonoids are widely distributed in plants, and many studies have shown that flavonoids possess antidiabetic properties and powerful antioxidant activity, which is an important factor in the treatment of diabetes and in preventing the development of diabetic angiopathies 6, 7. Rutin decreases the formation of reactive oxygen species, advanced glycation end-product precursors and inflammatory cytokines. These effects are considered to be responsible for the protective effect of rutin against hyperglycemia and dyslipidemia-induced nephropathy, neuropathy, liver damage, and cardiovascular disorders 8.
Modern pharmacotherapeutics, such as insulin, biguanides, sulfonylureas and thiazolidinediones, have emerged as a result of the advancement of modern medicine 9. The widely used allopathic oral hypoglycemic drugs for treating diabetes have been demonstrated to have a number of undesirable side effects. As a result, research is still concentrated on traditional medicinal plants that have been shown to have hypoglycemic effect; herbal therapy for diabetes is now a well-liked treatment option globally. Aims to formulate new pharmaceuticals with maximal performance and minimum adverse effects by extracting newer orally therapeutic hypoglycemic phytoconstituents. Diabetes therapy with herbs has been a focus of study on currently used traditional medicinal plants with established scientific evidence for hypoglycemic action.
Streptozotocin (STZ), a diabetogenic drug, is routinely administered to animals to cause diabetes. Diabetes mellitus can be triggered by 3 different strategies, including feeding rats a high-fat diet (HFD) and administering STZ injections after the administration of nicotinamide (STZ-NA) 10, 11. In order to evaluate the anti-diabetic effectiveness of Michelia champaca leaves, one of the 3 method was the STZ-NA model 12. The Magnoliaceae family includes Michelia champaca Linn, also known as Svarna Champa. The tree's unusual appearance, which includes a smooth trunk and large ovate, glossy leaves, is used in many conventional herbal remedies. It is well renowned for its gigantic, fragrant yellow blossoms. It flourishes from hot, humid weather and acidic soil 13. The leaves, stems, roots of Michelia champaca comprise a variety of phytoconstituents, including alkaloids, sugars, glycosides, tannins, proteins, amino acids and flavonoids 14.
MATERIALS AND METHODS:
Plant Material and Extract Preparation: The leaves of M. champaca L. were collected in and around Lucknow. Principal Scientist Dr. Lal Babu Chaudhary, certified and authenticated Michelia champaca Linn. leaves at Department of Plant Diversity, Systematics, and Herbarium Division, National Botanical Research Institute, Lucknow, Uttar Pradesh, India. The plant’s herbarium with accession number LWG 109567 has been deposited for future reference. Before being processed with a mechanical grinder, fresh leaves were dried in the shade. One kilogram of dried Michelia champaca leaf powder was extracted using petroleum ether, chloroform, ethyl acetate, ethanol, and distilled water in a Soxhlet apparatus. Collected extracts were heated to 40 °C in a hot air oven after being concentrated in a vacuum rotary evaporator at the Hygia Institute of Pharmaceutical Education and Research in Lucknow, Uttar Pradesh, India. The percentage yields of the ethanolic extract (12.5%)) and the aqueous extract (16.2%) of Michelia champaca leaves were calculated on the basis of the overall volume of powder used for the extraction.
Preliminary Phytochemical Screening: The extract was subjected to preliminary phytochemical screening for the identification of various active constituents by using standard procedure.
Drugs and Chemicals: Streptozotocin (Calbiochem, Mumbai, India), Nicotinamide (HiMedia, Mumbai, India), Glibenclamide pure powder (API as a gift sample from Sigma-Aldrich Ltd., Mumbai, India), and the Accu-chek Blood Glucose Test Strips and a functioning meter (Roche, India) were all acquired as chemicals and medications. All of the chemicals and reagents utilized were of the analytical sort.
Experimental Animals: Wistar rats of both sexes were provided by the Animal House at the United Institute of Pharmacy in Prayagraj, U. P., India. Standard husbandry practices were employed to house the animals, including a diurnal cycle at room temperature for 7-day acclimatization phase. The experimental investigation followed the standards for laboratory animal experimentation established by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), and the research protocol (UIP/IAEC/Nov.-2020/06) was approved by the United Institute of Pharmacy's Institutional Animal Ethics Committee (Reg. No. 1451/PO/Re/S/11/CPCSEA). For the experiment, animals of both sexes weighing 150–167 grams were employed.
Acute Toxicity Study of Extracts: According to the recommendations of the Organization for Environmental Control Development (OECD)-423, acute toxicity tests are carried out in experimental animals to obtain the LD50 value, which is used to estimate the therapeutic dose.
Different groups of rats received a single dosage of EEMC and AEMC extracts by oral gavage at doses of 5 mgkg-1, 50 mgkg-1, 300 mgkg-1, 2000 mgkg-1 and 5000 mgkg-1 in appropriate water (three each). Based on the LD50 calculation, dosages of 250 mgkg-1 and 500 mgkg-1 were selected for their ability to have an anti-diabetic effect.
Experimental Induction of Diabetes: Animals developed diabetes after exposure to the STZ-NA model. The trial involves repeated oral dosing over the course of 21 days. The overnight-fasting rats from Groups II to VII were given intraperitoneally single injected dose of NA (120 mg/kg body weight) and after 15 minutes, STZ (60 mg/kg body weight) intraperitoneally injected to induce Non-insulin-dependent diabetes mellitus.
Using a glucometer to measure the rats' blood sugar levels (BSL), was determined after 72 hours receiving STZ-NA injections. The animals were randomly assigned to seven different groups i.e. Group I to VII. Group I served as normal control (NC) containing 6 normal rats.
Experimental Approach for Anti-diabetic Activity: Six groups (6 animals in each group) of STZNAD rats of either sex were randomly created, and one group had six ND rats.
Throughout the trial, they had unrestricted access to water and animal food. For 21 days, oral doses of AEMC 250 mg/kg, AEMC 500 mg/kg, EEMC 250 mg/kg, and EEMC 500 mg/kg were administered.
Group I-NC: ND animals are given Distill water and served as Normal Control
Group II-DC: STZNAD animals used as Diabetic Control got Distill water.
Group III-STD: Glibenclamide 600 µg/kg/day given to STZNAD rats.
Group IV- AEMC 250mg/kg: STZNAD animals givenAEMC 250 mgkg-1day-1.
Group V- AEMC 500mg/kg: STZNAD animals given AEMC 500 mgkg-1 day-1.
Group VI- EEMC 250mg/kg: STZNAD rats received EEMC 250mg/kg /day.
Group VII- EEMC 500mg/kg: STZNAD rats received EEMC 500mg/kg /day.
Monitoring of Blood Sugar Level During Treatment: Animal's blood was drawn by clipping the tips of their tails, and blood sugar levels (BSL) were measured using an Accu-Check glucometer. Throughout the study period, BSL readings were taken every week at intervals (0th, 7th, 14th, and 21st days). The values of sample treated were compared with that of the standard group which was treated with Glibenclamide. Then, all animals were sacrificed by a sodium pentobarbitone overdose (150 mg/kg). The collected blood samples were immediately centrifuged at 2500 rpm for 15 min. The serum separated was collected in fresh serum tubes and stored tightly capped in refrigerator (2-4 degree centigrade). Effect of test extracts on antioxidant enzymes and lipid profile were assessed.
In-vivo Antioxidant Activity in Diabetic Rats: Measurement of Glutathione-S-transferase (GST), Reduced glutathione (GSH) and Glutathione peroxidase (GPx), Catalase (CAT), and Superoxide dismutase (SOD) were determined to ascertain its antioxidant activity by standard procedure 17.
Estimation of Biochemical Parameters: Measurement of lipid level, triglyceride and total cholesterol in blood serum were determined after 21 days.
Statistical Analysis: Mean ±Standard deviation were used to express the data. Utilizing the programme Graph Pad Prism 9.3.1, data analysis was carried out. All data were evaluated using a one-way Analysis of Variance (ANOVA), using Newman-Keuls test. It was thought that a value of p < 0.001 was more statistically significant.
RESULTS:
Preliminary Phytochemical Screening: Preliminary phytochemical screening revealed the presence of carbohydrate, phenolic compounds, sterols, alkaloids, glycoside and flavonoids in ethanolic and aqueous extract of leaves.
Study on the Acute Toxicity of Michelia champaca Leaf: At the maximum dose (5000 mg/kg), orally ingested AEMC and EEMC did not result in any deaths or adverse responses within 4 hours and after 14 days.
Effect of Michelia champaca Leaf Extracts on BSL: Table 1 Fig. 1, demonstrate the impact of oral daily dose of Michelia champaca aqueous extract and ethanolic extract (250 mg/kg and 500 mg/kg) on the blood glucose level over the course of 21 days. By the end of treatment, the blood glucose level reduced from 279.16 mg/dL to 113.16 mg/dL (59.46%) in rats treated at a dose of 250 mg/kg (p < 0.001) and blood glucose level reduced from 277.66 mg/ dL to 108.16 mg/dL (61.04%) in rats treated at a dose of 500 mg/kg (p<0.001) for the oral administration of aqueous extract. For the oral administration of 250 mg/kg of the ethanolic extract, the blood glucose level reduced from 276.83 mg/dL to 102.5 mg/dL (62.97%) at 21 days (p < 0.001) and blood glucose level reduced from 275.0 mg/ dL to 95.33 mg/dL (65.33%) in rats treated at a dose of 500 mg/kg (p<0.001) for the oral administration of ethanolic extract of Michelia champaca. For Glibenclamide (600 μg/kg), the blood glucose level reduced from 272.0 mg/dL to 92.16 mg/dL (66.11%) (p < 0.001. Orally ingested ethanolic extract of Michelia champaca leaves at a dose of 500 mg/kg (p<0.001) yields more significant results when compared to the hyperglycemia group.
TABLE 1: IMPACT OF VARIOUS EXTRACTS OF MICHELIA CHAMPACA LEAVES ON BLOOD GLUCOSE LEVEL OF STREPTOZOTOCIN NICOTINAMIDE CAUSED DIABETES IN RATS
Group(n) | 0 Day | 7th Day | 14th Day | 21st Day |
Normal Control | 80.5±6.97 | 81.0±8.85 | 81.83±9.15 | 82.33±9.58 |
Diabetic Control | 282.33±18.7z | 275.16±18.98z | 265.33±21.46z | 209.83±16.20z |
Standard Drug | 272.0±18.25 | 232.83±16.30 | 118.0±13.39 | 92.16±8.89c |
AEMC 250mg/kg | 279.16±18.72 | 253.16±18.34 | 142.66±16.71b | 113.16±6.30c |
AEMC 500mg/kg | 277.66±18.43 | 238.83±18.10 | 136.66±14.26c | 108.16±5.75c |
EEMC 250 mg/kg | 276.83±18.55 | 201.5±17.96a | 130.66±11.35c | 102.5±6.04c |
EEMC 500mg/kg | 275.0±18.29 | 195.33±17.15b | 124.0±13.13c | 95.33±6.42c |
The data represent as mean ± standard deviation of 6 animals in each group. bp<0.01 and cp < 0.001 compared to diabetic control group. zp< 0.001 as compared to control group.
FIG. 1: EFFECT OF AEMC AND EEMC ON BLOOD GLUCOSE LEVEL
Table 2 and Fig. 2 shows a significant increase in GSH, GST, GPx, and CAT and SOD in the Liver on the oral administration of Michelia champaca extracts at dose level 250 mg/kg and 500 mg/kg.
TABLE 2: THE EFFECT OF MICHELIA CHAMPACA EXTRACTS ON IN-VIVO ANTIOXIDANT ENZYMES OF LIVER
Group(n) | GSH | GST | GPx | CAT | SOD |
Normal Control (Group I) | 124.66±2.21 | 7.83±0.68 | 9.0±0.58 | 75.33±0.94 | 7.33±0.74 |
Diabetic Control(GroupII) | 71.66±1.97 | 5.5±0.95 | 6.67±0.94 | 37.33±1.10 | 4.16±0.68 |
Standard Drug(Group III) | 124±1.29c | 8.17±0.68c | 8.5±0.76c | 72.5±5.02 | 7.0±0.81c |
AEMC 250mg/kg(Group IV) | 113.83±2.67b | 4.83±0.68 | 4.83±0.68 | 68±3.41 | 5.66±0.74 |
AEPA500mg/kg (Group V) | 118.5±1.5 | 6±0.58 | 5.83±0.68 | 70.0±3.31 | 6.83±0.89 |
EEPA 250 mg/kg (Group VI) | 120.67±1.79 | 7.17±0.69 | 7±0.57 | 72.33±2.98 | 7.66±0.74 |
EEPA 500mg/kg (Group VII) | 122.5±1.70c | 8.33±0.75 | 8.33±0.74c | 74.66±2.80c | 8.66±0.74c |
The value represents mean ± SD for 6 rats per group. bp < 0.01 and cp < 0.001 compared to diabetic control group. zp< 0.001 compared versus normal control.
FIG. 2: EFFECT OF AEMC AND EEMC ON IN-VIVO ANTIOXIDANT ENZYMES IN LIVER
Table 3 and Fig. 3 shows a significant increase in GSH, GST, GPx, sand CAT in the kidney on the oral administration of Michelia champaca extracts at dose level 250 mg/kg and 500 mg/kg.
TABLE 3: THE EFFECT OF MICHELIA CHAMPACA EXTRACTS ON IN-VIVO ANTIOXIDANT ENZYMES OF KIDNEY
Group(n) | GSH | GST | GPx | CAT |
Normal Control (Group I) | 119±2.65 | 7.33±1.05 | 5.83±0.68 | 37.66±1.37 |
Diabetic Control(GroupII) | 47.33±1.79z | 4.83±1.07z | 3.5±0.5z | 22.66±1.59z |
Standard Drug(Group III) | 96.0±3.21c | 7.5±0.95c | 7.5±1.25c | 38.16±1.34c |
AEMC 250mg/kg(Group IV) | 88.35±2.35 | 4.5±0.95 | 4.16±0.68 | 27.83±1.06 |
AEPA500mg/kg (Group V) | 91.33±3.39b | 5.33±1.05b | 5.5±0.95b | 29.66±0.94 |
EEPA 250 mg/kg (Group VI) | 93.16±3.53c | 6.33±1.01c | 6.83±1.06 | 33.5±1.5c |
EEPA 500mg/kg (Group VII) | 96.33±2.49c | 7.5±0.96c | 7.83±1.06c | 37.16±1.57c |
The value represents mean ± SD for 6 rats per group. ap < 0.05, bp < 0.01 and cp < 0.001 compared to diabetic control group. zp< 0.001 compared versus normal control.
FIG. 3: EFFECT OF AEMC AND EEMC ON IN-VIVO ANTIOXIDANT ENZYMES IN KIDNEY
Table 4 and Fig. 4 demonstrate the triglyceride level in the STZ Nicotinamide-treated group was 71.17 mg/dL in the normal control group and 113.5 mg/dL in the diabetic control group (p < 0.001); the cholesterol level was 75.5 mg/dL in the normal control group and 149.67 mg/dL in the diabetic control group (p < 0.001); the HDL level was 52.16 mg/dL in the normal control group and 27.0 mg/dL in the diabetic control group (p < 0.001) and the LDL level was 31.66 mg/dL in the normal control group and 42.33 mg/dL in the diabetic control group (p < 0.001).
TABLE 4: IMPACT OF SEVERAL MICHELIA CHAMPACA EXTRACTS ON LIPID LEVEL, TRIGLYCERIDE AND TOTAL CHOLESTEROL IN BLOOD SERUM AFTER 21 DAYS
Category | Triglycerides | Cholesterol | HDL | LDL |
Normal Control | 71.17±5.39 | 75.5±4.19 | 52.16±2.91 | 31.66±4.92 |
Diabetic Control | 113.5±10.89z | 149.67±24.73z | 27.0±3.55z | 42.33±1.59z |
Standard Drug | 80.67±8.86c | 79.33±8.35c | 53.66±1.97c | 26.66±2.68c |
AEMC 250mg/kg | 105.67±11.33 | 99±11.46a | 39.0±2.94 | 38.16±1.95a |
AEMC 500mg/kg | 98.5±9.11b | 92±9.50b | 41.33±2.86b | 35.66±1.97 |
EEMC250 mg/kg | 91.17±8.13c | 85.16±8.78c | 47.0±2.44c | 32.33±3.09c |
EEMC 500mg/kg | 86.5±8.69c | 80.67±7.82c | 50.5±1.70c | 30.66±2.86c |
The value represents mean ± SD for six animals in each group. ap < 0.05, bp < 0.01 and cp < 0.001 compared to diabetic control group. zp< 0.001 compared versus normal control.
FIG. 4: EFFECT OF AEMC AND EEMC ON BIOCHEMICAL PARAMETERS
DISCUSSION: Diabetes is a severe, chronic condition that develops when the pancreas either generates insufficient amounts of insulin or when the body struggles to utilize the insulin that is produced. Diabetes prevalence has increased more quickly in low- and middle-income nations during the past ten years than in high-income nations. Type 2 diabetes affects the majority of persons who have the disease. This used to happen almost exclusively to adults, but today kids also experience it. All forms of diabetes raise the overall chance of premature death and can cause complications in various body parts. A heart attack, a stroke, kidney failure, eyesight loss, and nerve damage are all potential risks. Poorly managed diabetes during pregnancy raises the risk of complications, including foetal mortality. The combination of streptozotocin and nicotinamide in the type II diabetes model prevents pancreatic cells from becoming completely insulin-deficient. However, when nicotinamide is given first, it partially shields insulin-secreting pancreatic beta cells from the harmful effects of streptozotocin 18. Streptozotocin alone fully destroys pancreatic beta cells and renders them insulin-deficient. Secondary metabolites such as alkaloids, flavonoids, and saponins reduced plasma cholesterol and triglycerides significantly, significantly decreased glucose levels and significantly increased hepatic glucokinase activity. This was most likely accomplished by increasing the release of insulin from pancreatic islets 19. Increased enzymatic antioxidant activity was seen after treatment with M. champaca, which may be related to flavonoids. Due to their well-known antioxidant qualities, the flavonoids in the leaf extract scavenge the free radicals produced during diabetes. All the above observations suggest that M. champaca can be a promising antidiabetic and antioxidant drug.
CONCLUSION: We concluded that Michelia champaca extracts had anti-diabetic properties and significantly reduced blood glucose levels. It has been established that Michelia champaca is a reliable diabetes medication. It is possible that the observed antihyperglycemic activity of Michelia champaca leaves is related to the presence of flavonoids, sterols, phenols, glycosides, and alkaloids as active constituents. The current analysis has also paved the way for additional study, particularly in relation to the creation of effective herbal remedies for diabetes mellitus using Michelia champaca leaves.
ACKNOWLEDGEMENT: The authors are acknowledging Dr. Alok Mukerjee, Professor, United Institute of Pharmacy, Prayagraj, U.P., India, for providing facilities to carry out the pharmacological activity.
CONFLICTS OF INTEREST: The authors declare no conflicts of interest.
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How to cite this article:
Nanda J, Mani M, Mishra SB and Verma N: Michelia champaca leaf extracts exhibit hypoglycemic effect and hypolipidemic activity in streptozotocin-nicotinamide-induced diabetic rats. Int J Pharm Sci & Res 2022; 13(12): 5200-06. doi: 10.13040/IJPSR.0975-8232.13(12).5200-06.
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IJPSR
Jyoti Nanda *, Munesh Mani , Shanti Bhushan Mishra and Neeraj Verma
School of Pharmaceutical Sciences, CSJM University, Kanpur, Uttar Pradesh, India.
jyotinanda954@gmail.com
14 September 2022
11 November 2022
15 November 2022
10.13040/IJPSR.0975-8232.13(12).5200-06
01 December 2022