TOXICITY STUDIES OF A SAPONIN ISOLATED FROM THE FRUITS OF MOMORDICA DIOICA IN RATS
HTML Full TextTOXICITY STUDIES OF A SAPONIN ISOLATED FROM THE FRUITS OF MOMORDICA DIOICA IN RATS
Deepak Kumar Jha *, Raju Koneri and Suman Samaddar
Department of Pharmacology, Karnataka College of Pharmacy, Bangalore - 560064, Karnataka, India.
ABSTRACT: The objective of this study was to evaluate the toxicity of a saponin isolated from the fruits of the Momordica dioica as per OECD guidelines. Saponin was isolated from the methanolic extract of the fruits of M. dioica, and acute, sub-acute, sub-chronic and chronic toxicity was evaluated. The study of the acute toxicity of saponin M. dioica (SMD) at a single dose of 5000 mg/kg/b.w. by oral route in, recorded no mortality, and showed no change in the general behavior of the treated rats for 14 days. Administration at the doses; 1000, 500, and 250 mg/kg/b.w. by oral route in rats for 28 days and an animal showed doses of 500 and 250 mg/kg/b.w. of SMD was safe, no mortality for sub-acute but in a repeated dose of 1000 mg/kg/b.w. group produced treatment-related signs of toxicity. The study of Sub-chronic toxicity at the doses; 500, 250 and 100 mg/kg/b.w. of SMD in rats for 90 days and animals showed normal; hematological and biochemical parameters and gross findings compared to the control group. Histopathology examinations of all vital organs did not reveal morphological alteration especially the dose of 250 and 100 mg/kg/b.w. This finding further confirmation by the administration of extract of SMD 250, 100 and 50 mg/kg/b.w. for 180 days produced no clinical signs for toxicity or mortality. We may conclude that SMD is not toxic in the doses of 250, 100 and 50 mg/kg/b.w and did not produce any toxic signs or any remarkable histopathological signs or serum chemical alteration.
Keywords: |
Momordica dioica, Saponin, Acute, Sub-acute, Sub-chronic, and Chronic toxicity
INTRODUCTION: Medicinal plants, either as an extract, pure compound or as a derivative, offer unlimited opportunities for the discovery of new drugs. Use of herbal medicines represents a long history of human interactions with the environment. Plants used for traditional medicine contain a wide range of substances that can be used to treat chronic as well as infectious diseases. However, there is little information or evidence available concerning the possible toxicity that medicinal plants may cause to the consumers 1.
Several modern drugs currently in use have been obtained through medicinal plants. Despite the profound therapeutic advantages possessed by some of the medicinal plants, some constituents of medicinal plants are potentially toxic, mutagenic, carcinogenic and teratogenic. This raises a concern about the potentially toxic effects resulting from the short-term and long-term use of such medicinal plants.
Therefore, evaluating the toxicological effects of any medicinal plant extract intended to be used in animals or humans is a crucial part of its assessment for potential toxic effects. Momordica dioica is a perennial, dioecious; cucurbitaceous climbing creeper (commonly known as kakrol, spiny gourd or teasle gourd) plant is used both in the prevention and cure of various diseases and in the food of humans.
It is necessary that we should have full knowledge regarding the therapeutically use and pharmacology activities for their proper utilization 2. The fruits of Momordica dioica used to management of diabetes and other health problems like; antimicrobial activity, antibacterial activity and fruits could be a good supplement for some nutrients, minerals, and fatties such as fiber, protein, carbohydrate, calcium, magnesium, oleic acid, and linoleic acid. The fruits could be promoted as a mineral and vitamin.3, 4 Based on Momordica dioica use in traditional practices and the literature references, the present study was undertaken to evaluate the comprehensive acute, sub-acute, sub-chronic and chronic toxicity in rats and is reported hereunder.
MATERIALS AND METHODS
Extraction and Isolation of Saponin: The fruits of Momordica dioica were collected during June month, Bangalore, Karnataka. The sample was identified by Botanist Dr. Geetanjali department of botany, Sree Siddaganga College, Tumkur University (Reference No: 141/17-18). The fruits of Momordica dioica were, chopped into small pieces and dried under shade at room temperature. The dried fruits were powdered and passed through the sieve (coarse10/40). The powder was used for the preparation of the methanolic extract. Each 100 g powder was subjected to extraction with 1000 ml methanol in a reflux condenser for 3 cycles of 7 hrs each till the volume reduced to half. The extract was filtered through Whatman filter paper no.1 and evaporated to dryness to get constant weight. Five grams of methanolic extract of Momordica dioica was subjected to saponification in a 500 ml conical flask and hydrolyzed with 100 ml of 0.5 N potassium hydroxide for 1-2 h. The whole mixtures were diluted with water and extracted with diethyl ether. The ether layers were evaporated by water bath further isolation and purification was achieved through fractionation method of TLC that yielded a pure saponin fraction and the residues were tested for the confirmation of steroidal saponin 5-8.
Animals: Wistar rats (both sex, 5-6 weeks old) weighing 190 ± 10 g were used for the experiment. They were acclimatized for one week before experiment. Animals were caged in fully ventilated room, were maintained in 12:12 h light and dark cycle and were housed at a temperature of 25 ± 2 °C. They had free access to a standard chow diet and water ad libitum. All the experiments conducted on the animals were by the standards set for the use of the laboratory animal use, and the experimental protocols were duly approved by the IAEC of Karnataka College of Pharmacy, Bangalore (Reg. no: KCP-IAEC2/17-18/15-04-07). The entire study was conducted by the CPCSEA guidelines.
Experimental Design: The Animals toxicity studies were conducted in compliance with OECD-guideline. Animals were observed individually at least once during the first 30 min after dosing, periodically during the first 24 h (with special attention given during the first 4 h), and daily thereafter; For acute toxicity total of 14 days (OCED-425), 28 days (OCED-407, for Subacute toxicity), 90 days (OCED-408, for Sub-chronic toxicity) and 180 days (OCED-452, for chronic toxicity). The times at which signs of toxicity appear and disappear are important, especially if there is a tendency for toxic signs to be delayed.
All observations were systematically recorded with individual records being maintained for each animal. The animal was observed closely for any behavioral changes, body weight changes and mortality and later withdrawal the blood for hematological and biochemical investigations from cardiac puncture by gave light sedation (low dose of pentobarbital) and then sacrificed the animal with a high dose of pentobarbital for tissue histology.
Biochemical Parameters: The biochemical parameters like Glucose (Using digital glucometer; one-touch select, LifeScan Scotland Ltd, UK), Triglycerides, Total Cholesterol, LDL, VLDL, and HDL were estimated as per the standard procedure prescribed by the manufacturer’s instruction manual provided in the kit. (DELTA LABS kit, Bangalore, India) using Semi-Auto analyzer including serum levels of total proteins (TP), bilirubin, alkaline phosphate (ALP), creatinine and albumin (ALB). Serum electrolytes the blood samples were centrifuged at 3000 rpm for 15 min. Diagnostic kits were used to evaluate Sodium, Potassium, and chloride.
Hematological Parameters: Haematological analysis of the blood samples was performed using an automatic hematology analyzer. The parameters which were evaluated included: red blood cells (RBC) count; hemoglobin (Hb); platelets (PLT); leukocytes (WBC) count; and neutrophils, lymphocytes.
Histopathological Study: After sacrificing the rats, parts of the liver, kidney, lung, heart, and spleen tissues were collected for histological studies. The tissues were washed in normal saline and fixed immediately in 10% formalin for a period of at least 24 h, dehydrated with alcohol, embedded in paraffin, cut into 4-5 m thick sections, and stained with hematoxylin-eosin dye for photomicroscopic observation. The microscopic features of the organs of rats were compared with the control group.
Statistical Analysis: The results are expressed as Mean ± SD in each group. The significance of difference among the groups was assessed using one-way ANOVA followed by Tukey’s multiple comparison tests (Graph Pad Prism v5.0).
RESULTS:
Assessment of Acute Toxicity of Phyto-constituent in Animal: In the acute oral toxicity study, the isolated saponin Momordica dioica (SMD) from methanolic extract of fruits of Momordica dioica was found to be safe up to a dose of 5000 mg/kg without any mortality.
TABLE 1: EFFECT OF ORAL ADMINISTRATION OF SMD 5000 mg/kg ON BODY WEIGHT
Group | 0 days | 7 days | 14 days |
Normal control | 200±1.23 | 201±1.18 | 203±1.28 |
SMD 5000 mg/kg | 203±1.28*** | 205±0.70*** | 206±0.149*** |
Values are expressed as Mean ± S.E.M (n=6). ***P<0.05 when compared with normal control after 14 days. The body weights of normal and treated rats with a dose of 5000 mg/kg of SMD and data didn’t show much variation between the groups even after 14 days.
GRAPH 1: EFFECT OF ORAL ADMINISTRATION OF SMD 5000 mg/kg ON PLASMA GLUCOSE LEVELS AND LIPID PROFILES. Values are expressed as Mean ± S.E.M (n=6). Examination of plasma glucose and Triglycerides (TG), VLDL did not show much variation between the treated groups and their respective control groups (***P<0.05) after 14 days. But there were some significant variation (Graph 01) between TC, HDL and LDL concentrations in the treated groups when we were compared those in the normal control groups (**P<0.05) after 14 days.
TABLE 2: EFFECT OF ORAL ADMINISTRATION OF SMD 5000 mg/kg ON SERUM BIOCHEMICAL PARAMETERS LEVELS
Group/Test Parameters | Total Protein (TP) gm/dl | Albumin (gm/dl) | Globulin (gm/dl) |
Normal Control | 5.5±1.229 | 3±1.033 | 2.5±0.6708 |
SMD 5000 mg/kg | 10.9±1.515** | 3.9±0.8563*** | 7±1.39** |
Values are expressed as Mean ± S.E.M (n=6). ***P<0.05, **P<0.05 when compared with normal control after 14 days. Total protein and globulin were significantly increased in treated groups P<0.05. Moreover, a mild increased in serum albumin was observed in the treated rats with SMD 5000 mg/kg compared with their respective control group.
Group/Test Parameters | Total Bilirubin (mg/dl) | Direct Bilirubin (mg/dl) | Blood Urea (mg/dl) | Serum Creatinine (mg/dl) | Uric acid
(mg/dl) |
Normal Control | 0.6±0.9888 | 0.1±0.8333 | 28±0.1384 | 0.7±0.1476 | 2.6±1.033 |
SMD 5000 mg/kg | 1.5±0.654** | 1±0.0355** | 43±0.7746** | 0.5±0.2301*** | 16.1±0.5774** |
Bilirubin, blood urea and uric acid were significantly increased in treated groups were observed P<0.05. Moreover, a mild increase in serum creatinine was observed P<0.05 in the rats treated with SMD 5000 mg/kg compared with their respective control group.
Group/Test Parameters | Sodium
(mmol/L) |
Potassium
(mmol/L) |
Chloride
(mmol/L) |
Alkaline Phosphate
(ALP) U/L |
Normal Control | 140±0.1478 | 4±0.2124 | 101±0.4597 | 151±1.145 |
SMD 5000 mg/kg | 138±0.70*** | 4.2±0.7638*** | 102±0.8333*** | 41±1.232** |
Examination of electrolytes; Sodium, Potassium, and Chloride didn’t show much variation between the treated groups P<0.05 and their respective control but there is significant decreased P<0.05 alkaline phosphate from those in the normal control group.
TABLE 3: EFFECT OF ORAL ADMINISTRATION OF SMD 5000 mg/kg ON HAEMATOLOGICAL LEVELS
Parameters/Groups | Normal control | 5000 mg/kg of SMD |
Hb (gm/dl) | 14.2±1.14 | 14.15±1.28*** |
WBC (c/cmm) | 8,600±0.2124 | 7,338±1.18*** |
RBC (m/cmm) | 8.5±0.4597 | 8.6±0.5746*** |
Neutrophils (%) | 60±0.2301 | 55±0.5774*** |
Lymphocyte (%) | 35±0.1476 | 32±0.6667*** |
Platelet (lakh/cmm) | 3.22±1.14 | 2.22±1.49*** |
Values are expressed as Mean ± S.E.M (n=6). ***P<0.05 when compared with normal control after 14 days. Examination of hematological levels did not show much variation between the treated groups and their respective control groups.
Histological Examination of Haematoxylin and Eosin (H&E) Stained Tissues: Histopathological examinations were performed on the different organs to assess whether or not organs or tissues had been damaged. Group Normal- Liver showing normal architecture of hepatocytes arranged cord like manner around the central vein with cytoplasm staining red and vesicular type of nucleus. Spleen showed the white pulp with lymphocytes and red pulp with red blood cells with normal architecture. The lung tissue has the appearance of fine lace because most of the lung is composed of thin-walled alveoli and composed of a single layer of connective tissue. Bronchioles can be recognized by the fact that they are lined by ciliated columnar epithelium.
Normal findings in alveoli: alveolar macrophages, corpora amylacea, blue bodies, and mega-karyocytes. Kidney showed normal architecture of glomerulus with a tuft of capillaries surrounded by Bowman’s capsule lined by columnar epithelial cells. Heart showing normal striated skeletal muscles with oval-shaped nucleus. Group SMD 5000 mg/kg- liver showed normal architecture but vascular congestion, feathery degeneration of hepatocytes with unstained vacuoles inside, indicating loss of cytoplasm with a condensed nucleus and focal portal inflammation. Spleen showed congested red pulp. Lung showed focally collapsed alveoli, focal septal inflammation. Kidney and Heart were showed no observable histological changes.
FIG. 1: EFFECT OF ORAL ADMINISTRATION OF SMD 5000 mg/kg ON THE DIFFERENT ORGANS COMPARE TO NORMAL CONTROL AFTER 14 DAYS
Assessment of Sub Acute Toxicity of Phytoconstituent in Male and Female Rats:
TABLE 4: EFFECT OF ORAL 28 DAYS OF REPEATED ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON BODY WEIGHT
Group | Effect of oral administration of SMD on body weight (gm) | |||||
0 days | 7 days | 14 days | 21 days | 28 days | ||
Male | Male control group | 200±1.0245 | 204±1.0085 | 205±1.5982 | 205±1.0049 | 206±1.0087 |
SMD 1000 mg/kg | 201±1.0064*** | 199±1.0256*** | 199±1.0095*** | 195±0.0932*** | 195±0.8924*** | |
SMD 500 mg/kg | 195±1.0023*** | 193±1.0325*** | 192±1.0068*** | 190±0.0891*** | 189±1.2565*** | |
SMD 250 mg/kg | 190±0.9621*** | 189±1.0269*** | 188±1.0053*** | 185±0.0856*** | 184±1.2762*** | |
Female | Female control group | 198±1.1240 | 200±1.4785 | 201±1.2455 | 201±1.2454 | 202±1.2544 |
SMD 1000 mg/kg | 199±1.0045### | 196±1.0531### | 195±1.008### | 192±0.0873### | 191±1.0056### | |
SMD 500 mg/kg | 195±1.0017### | 192±1.0432### | 192±0.0856### | 190±0.0885### | 189±1.2475### | |
SMD 250 mg/kg | 203±1.0056### | 200±0.0654### | 201±1.2545### | 199±1.0465### | 197±1.2453### |
Values are expressed as Mean ± S.E.M (n=5). ***P<0.05 when compared with repeated administration of SMD Male rats group concerning the male control group, ###P<0.05 when compared with repeated administration of SMD Female rats group concerning the female group. The body weights of repeated oral administration rats with dose of SMD were displayed in Table 4 for the male and the female rats respectively, data didn’t show much variation between the groups with respective control group even after 28 days however there were some mild changes of repeated oral dose of SMD 1000 mg/kg and 500 mg/kg observed within the groups.
GRAPH 2: EFFECT OF ORAL ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON PLASMA GLUCOSE LEVELS AND LIPID PROFILES. Values are expressed as Mean ± S.E.M (n=5). Examination of plasma glucose did not show much variation between the treated groups and their respective control groups (P<0.05) after 28 days. But there was some significant variation between TG, TC, HDL, LDL and VLDL concentrations in the treated groups (graph 02) when we were compared those in the normal control groups (P<0.05) after 28 days.
TABLE 5: EFFECT OF ORAL ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON SERUM TOTAL PROTEIN, ALBUMIN AND GLOBULIN
Group | Total Protein (TP) gm/dl | Albumin gm/dl | Globulin gm/dl | |
Male | Male control group | 5.5±0.76 | 3±0.87 | 2.5±1.24 |
SMD 1000 mg/kg | 8.5±0.58** | 3.5±1.14*** | 5±0.47** | |
SMD 500 mg/kg | 7.9±0.68** | 3.2±1.49*** | 4.7±0.21** | |
SMD 250 mg/kg | 7±0.76** | 3.3±0.60*** | 4±0.63** | |
Female | Female control group | 5.4±0.45 | 3.1±0.42 | 2.2±0.57 |
SMD 1000 mg/kg | 8.1±0.48## | 3.7±0.95### | 2.6±1.35### | |
SMD 500 mg/kg | 7.8±0.46## | 3.4±1.68### | 5.1±1.67## | |
SMD 250 mg/kg | 5.7±0.64### | 3.2±0.16### | 4.8±0.67## |
Values are expressed as Mean ± S.E.M (n=5). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 28 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 28 days. Total protein and globulin were significantly increased in treated groups were observed P<0.05. Moreover, a mild increase in serum albumin was observed in the rats (both male and female) treated with 1000 mg/kg compared with their respective control group.
TABLE 6: EFFECT OF ORAL ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON SERUM TOTAL BILIRUBIN, DIRECT BILIRUBIN, BLOOD UREA, CREATININE, AND URIC ACID
Group | Bilirubin total mg/dl | Bilirubin direct mg/dl | Blood Urea mg/dl | Serum Creatinine mg/dl | Uric acid mg/dl | |
Male | Male control group | 0.6±0.70 | 0.1±0.14 | 28±1.32 | 0.7±0.24 | 2.6±0.24 |
SMD 1000 mg/kg | 0.8±0.18** | 0.5±0.52** | 80±1.25** | 0.5±0.58*** | 15±0.65** | |
SMD 500 mg/kg | 0.5±1.28*** | 0.3±0.63*** | 71±1.24** | 0.3±0.95*** | 12.6±0.45** | |
SMD 250 mg/kg | 0.2±1.23*** | 0.2±0.65*** | 28±1.26*** | 0.3±0.78*** | 8±0.54** | |
Female | Female control group | 0.6±0.57 | 0.1±0.45 | 30±0.15 | 0.5±0.65 | 2.7±0.65 |
SMD 1000 mg/kg | 0.9±1.17## | 0.5±1.25## | 89±0.17## | 0.4±0.57### | 16.2±0.87## | |
SMD 500 mg/kg | 0.4±0.15### | 0.2±1.65### | 69±0.89## | 0.6±0.68### | 13.4±1.15## | |
SMD 250 mg/kg | 0.3±0.17### | 0.1±1.78### | 27±0.65### | 0.3±0.64### | 8.9±1.65## |
Values are expressed as Mean ± S.E.M (n=5), ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 28 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 28 days. Blood urea and uric acid were significantly increased in treated groups were observed P<0.05. Moreover, a mild increase in bilirubin was observed P<0.05 in the rats treated with 1000 mg/kg compared with their respective control group.
TABLE 7: EFFECT OF ORAL ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON SERUM SODIUM, POTASSIUM, CHLORIDE, AND ALKALINE PHOSPHATE
Group | Sodium
(mmol/L) |
Potassium (mmol/L) | Chloride (mmol/L) | Alkaline Phosphate (ALP) U/L | |
Male | Male control group | 140±0.44 | 4±1.22 | 101±0.98 | 151±0.02 |
SMD 1000 mg/kg | 136±1.08*** | 3.7±1.69*** | 97±0.22*** | 289±0.03** | |
SMD 500 mg/kg | 130±1.47*** | 3.5±0.66*** | 95±0.35*** | 148±0.76*** | |
SMD 250 mg/kg | 139±0.98*** | 3.1±1.03*** | 90±0.09*** | 150±0.92*** | |
Female | Female control group | 139±0.79 | 3.9±0.67 | 103±0.42 | 156±0.82 |
SMD 1000 mg/kg | 136±1.30### | 3.9±0.57### | 95±0.65### | 249±0.75## | |
SMD 500 mg/kg | 131±1.17### | 3.4±0.77### | 99±0.85### | 159±1.32### | |
SMD 250 mg/kg | 138±0.77### | 3.2±1.91### | 102±1.23### | 147±0.54### |
Values are expressed as Mean ± S.E.M (n=5). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 28 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 28 days. Examination of electrolytes; Sodium, Potassium, and Chloride didn’t show much variation between the treated groups P<0.05 and their respective control but there is significant increased P<0.05 alkaline phosphate dose of SMD 1000 mg/kg with their respective normal control group.
TABLE 8: EFFECT OF ORAL ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON HAEMATOLOGICAL LEVELS
Group | Hb
(gm/dl) |
WBC (c/cmm) | RBC (m/cmm) | Neutrophils (%) | Lymphocyte (%) | Platelet (lakh/cmm) | |
Male | Male control group | 16±1.28 | 8.600±0.21 | 8.5±0.45 | 60±0.23 | 35±0.14 | 3.22±1.14 |
SMD
1000 mg/kg |
9.1±1.14** | 7.545±0.60** | 9.9±1.01** | 72±0.25** | 17±1.02** | 6.81±0.35** | |
SMD
500 mg/kg |
10.2±1.28** | 8.000±0.17*** | 8.9±1.25*** | 65±1.54*** | 32±1.35*** | 3.67±1.14*** | |
SMD
250 mg/kg |
13.9±1.18*** | 8.500±0.23*** | 8.4±1.15*** | 60±1.35*** | 30±1.65*** | 2.81±0.36*** | |
Female | Female control group | 15±1.13 | 8.500±1.13 | 8.5±0.74 | 61±1.33 | 34±1.45 | 3.25±0.35 |
SMD 1000 mg/kg | 8.2±1.17## | 7.254±1.68## | 10.7±0.84## | 67±0.98## | 19±1.35## | 6.89±0.37## | |
SMD 500 mg/kg | 10.4±0.13## | 8.400±1.49### | 9.2±0.57### | 6±0.64### | 34±1.54### | 3.68±1.31### | |
SMD 250 mg/kg | 12.1±1.43### | 8.600±0.69### | 8.6±0.68### | 58±0.68### | 26±1.35### | 2.87±1.12### |
Values are expressed as Mean ± S.E.M (n=5). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 28 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 28 days. Examination of hematological levels did not show much variation between the treated groups and their respective control groups. But some mild changes got observed especially in Hb, WBC, Lymphocytes, and platelets on a higher dose of SMD i.e. 1000 mg/kg with their respective control group.
Histological Examination of Haematoxylin and Eosin (H&E) Stained Tissues:
FIG. 2: EFFECT OF ORAL ADMINISTRATION OF SMD 1000 mg/kg, 500 mg/kg AND 250 mg/kg ON THE DIFFERENT ORGANS OF MALE RATS AND THE FEMALE RATS RESPECTIVELY AFTER 28 DAYS. The histological features of the different organs were displayed in the figure for the male and the female rats, respectively. SMD 1000 mg/kg Liver- the morphology of the hepatic cells in both the male and the female groups was normal. However, mild vascular degeneration with the condensed nucleus of hepatocytes. SMD 500 mg/kg- showed mild necrosis and 250 mg/kg- showed no observable changes. SMD 1000 mg/kg Spleen- had shown normal architecture, normal white pulp but congested red pulp. SMD 500 mg/kg and 250 mg/kg were showed no observable histological changes in both the male and the female group. SMD 1000 mg/kg Lung- Showed Peribronchial cuffing due to the inflammation, thickening of the bronchial wall were observed in both the male and female groups. SMD 500 mg/kg showed focally collapsed alveoli, and SMD 250 mg/kg showed normal alveoli in both male and the female groups. Kidney and Heart were showed no observable histological changes in all groups.
Assessment of subchronic toxicity of phytoconstituent in Male and Female rats:
TABLE 9: EFFECT OF ORAL 90 DAYS OF REPEATED ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON BODY WEIGHT
Group | 0 days | 15 days | 30 days | 45 days | 60 days | 75 days | 90 days | |
Male | Male control group | 201±0.021 | 202.5±0.245 | 207±1.154 | 209±0.245 | 210±0.031 | 215±0.214 | 214±0.134*** |
SMD
500 mg/kg |
201±0.013*** | 199±0.749*** | 199±0.014*** | 195±1.138*** | 195±0.211*** | 192±0.545** | 190±0.872** | |
SMD
250 mg/kg |
194±0.011*** | 194.5±0.070*** | 192±1.245*** | 190±0.065*** | 189±0.745*** | 189.5±0.354*** | 191±0.748*** | |
SMD
100 mg/kg |
190±1.131*** | 189±0.749*** | 191±1.235*** | 194±0.139*** | 194.5±1.745*** | 196±0.564*** | 197±0.345*** | |
Female | Female control group | 198±0.020 | 200±1.138 | 201±0.006 | 201±0.024 | 202±1.366 | 207±1.344 | 209±0.545 |
SMD
500 mg/kg |
199±0.024### | 196±0.872### | 195±0.094### | 192±0.048### | 191±1.399### | 189±0.354## | 187±0.254## | |
SMD
250 mg/kg |
195±0.749### | 192±1.078### | 192±0.654### | 190±0.744### | 189±1.775### | 191.5±0.365### | 190±0.545### | |
SMD
100 mg/kg |
200±0.015### | 200.5±0.872### | 201±0.261### | 203±0.007### | 203.5±0.945### | 205±0.687### | 206.5±0.655### |
Values are expressed as Mean ± S.E.M (n=10). ***P<0.05 & **P<0.05 when compared with repeated administration of SMD Male rats group concerning the male control group, ###P<0.05 & ##P<0.05 when compared with repeated administration of SMD Female rats group concerning the female group. The body weights of repeated oral administration rats with dose of SMD were displayed on Table for the male and the female rats respectively, data didn’t show much variation between the groups with respective control group even after 90 days however there were some mild changes after the two months of repeated oral dose of SMD 500 mg/kg observed within the male and the female groups.
GRAPH 3: EFFECT OF ORAL ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON PLASMA GLUCOSE LEVELS AND LIPID PROFILES. Values are expressed as Mean ± S.E.M (n=10). Examination of plasma glucose did not show much variation (P<0.05) between the treated groups even after 90 days and their respective control groups (graph 03). But both the group were showed significant variation (P<0.05) between TG, TC, HDL, LDL and VLDL concentrations in the treated groups when we were compared those in the normal control group.
TABLE 10: EFFECT OF ORAL ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON SERUM TOTAL PROTEIN, ALBUMIN AND GLOBULIN
Group | Total Protein (TP) gm/dl | Albumin gm/dl | Globulin gm/dl | |
Male | Male control group | 5.5±0.15 | 3±0.21 | 2.5±0.32 |
SMD 500 mg/kg | 8.6±0.46** | 3.9±0.23** | 5.5±0.64** | |
SMD 250 mg/kg | 7.1±0.14** | 3.2±0.22*** | 4.8±0.54** | |
SMD 100 mg/kg | 6±0.54*** | 3.1±0.11*** | 2.9±0.51*** | |
Female | Female control group | 5.4±0.23 | 3.1±0.35 | 2.2±0.15 |
SMD 500 mg/kg | 8.2±0.31## | 4.1±068## | 5.1±0.34## | |
SMD 250 mg/kg | 7.6±0.24## | 3.8±0.45### | 4.9±0.64## | |
SMD 100 mg/kg | 5.7±0.12### | 3.2±0.54### | 3±0.58### |
Values are expressed as Mean ± S.E.M (n=10). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 90 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 90 days. Total protein and globulin were significantly increased in treated groups were observed P<0.05 (Table 10). Moreover, a mild increase in serum albumin was observed in the rats treated with 500 mg/kg compared with their respective control group.
TABLE 11: EFFECT OF ORAL ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON SERUM TOTAL BILIRUBIN, DIRECT BILIRUBIN, BLOOD UREA, CREATININE, AND URIC ACID
Group | Bilirubin total mg/dl | Bilirubin direct mg/dl | Blood Urea mg/dl | Serum Creatinine mg/dl | Uric acid mg/dl | |
Male | Male control group | 0.6±0.39 | 0.1±0.29 | 28±0.39 | 0.7±0.33 | 2.6±0.32 |
SMD 500 mg/kg | 0.8±0.42*** | 0.5±0.11*** | 80±0.64** | 0.5±0.31*** | 15±0.51** | |
SMD 250 mg/kg | 0.5±0.39*** | 0.3±0.41*** | 31±0.45*** | 0.3±0.39*** | 7.6±0.25** | |
SMD 100 mg/kg | 0.2±0.27*** | 0.2±0.75*** | 28±0.51*** | 0.3±0.36*** | 3.3±0.55*** | |
Female | Female control group | 0.6±0.12 | 0.1±0.21 | 30±0.21 | 0.5±0.65 | 2.7±0.42 |
SMD 500 mg/kg | 0.9±0.16### | 0.5±0.39### | 99±0.75## | 0.7±0.55### | 16.3±0.24## | |
SMD 250 mg/kg | 0.7±0.51### | 0.3±0.51### | 34±0.45### | 0.6±0.44### | 7.2±0.12## | |
SMD 100 mg/kg | 0.4±0.61### | 0.2±0.64### | 31±0.37### | 0.3±0.65### | 3.4±0.24### |
Values are expressed as Mean ± S.E.M (n=10). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 90 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 90 days. Blood urea and uric acid were significantly increased in treated groups were observed P<0.05. Moreover, a mild increase in bilirubin was observed P<0.05 in the rats treated with 500 mg/kg compared with their respective control group.
TABLE 12: EFFECT OF ORAL ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON SERUM SODIUM, POTASSIUM, CHLORIDE, AND ALKALINE PHOSPHATE
Group | Sodium
(mmol/L) |
Potassium (mmol/L) | Chloride (mmol/L) | Alkaline Phosphate (ALP) U/L | |
Male | Male control group | 140±0.028 | 4±0.034 | 101±0.101 | 151±0.351 |
SMD 500 mg/kg | 139±0.045*** | 3.4±0.056*** | 97±0.087*** | 179±0.341** | |
SMD 250 mg/kg | 141±0.135*** | 3±0.060*** | 99±0.171*** | 148±0.042*** | |
SMD 100 mg/kg | 140±0.341*** | 3.9±0.046*** | 91±0.033*** | 149±0.058*** | |
Female | Female control group | 139±0.093 | 3.9±0.076 | 103±0.034 | 156±0.082 |
SMD 500 mg/kg | 136±0.050### | 3.9±0.217### | 95±0.044### | 184±0.028## | |
SMD 250 mg/kg | 130±0.037### | 3.4±0.172### | 99±0.104### | 159±0.011### | |
SMD 100 mg/kg | 141±0.021### | 3.2±0.245### | 102±0.084### | 147±0.211### |
Values are expressed as Mean ± S.E.M (n=10). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 90 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 90 days. Examination of electrolytes; Sodium, Potassium, and Chloride didn’t show much variation between the treated groups P<0.05 and their respective control but there were mild changes P<0.05 in alkaline phosphate dose of SMD 500 mg/kg with their respective normal control group.
TABLE 13: EFFECT OF ORAL ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON HAEMATOLOGICAL PARAMETERS
Group | Hb (gm/dl) | WBC (c/cmm) | RBC (m/cmm) | Neutrophils (%) | Lymphocyte (%) | Platelet (lakh/cmm) | |
Male | Male
control group |
16±1.01 | 8.600±0.75 | 8.5±0.97 | 60±0.01 | 35±0.16 | 3.22±1.14 |
SMD
500 mg/kg |
10.12±0.65** | 7.337±0.06** | 10.9±0.71** | 61±0.14*** | 19±0.33** | 8.97±1.24** | |
SMD
250 mg/kg |
14.12±0.78*** | 8.000±0.65*** | 9.92±0.73*** | 54±0.25*** | 21±0.21** | 3.67±1.39*** | |
SMD
100 mg/kg |
15.9±1.30*** | 8.500±0.92*** | 9.47±0.94*** | 59±1.24*** | 30±1.34*** | 2.81±0.25*** | |
Female | Female control group | ||||||
SMD
500 mg/kg |
14.9±0.06 | 8.654±0.31 | 8.6±0.36 | 58±0.20 | 31±1.97 | 3.12±1.33 | |
SMD
250 mg/kg |
10.2±0.63## | 7.355±0.96## | 10.7±0.24## | 60±1.93### | 16±1.64## | 7.98±1.54## | |
SMD
100 mg/kg |
14.1±1.21### | 8.354±1.91### | 9.21±0.92### | 50±0.63### | 19±0.58## | 3.54±1.66### |
Values are expressed as Mean ± S.E.M (n=10). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 90 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 90 days. Examination of hematological levels did not show much variation between the treated groups and their respective control groups. But some mild changes got observed especially in Hb, WBC, Lymphocytes, and platelets on SMD 500 mg/kg (P<0.05) with their respective control group.
Histological Examination of Haematoxylin and Eosin (H&E) Stained Tissues:
FIG. 3: EFFECT OF ORAL ADMINISTRATION OF SMD 500 mg/kg, 250 mg/kg AND 100 mg/kg ON THE DIFFERENT ORGANS OF MALE AND THE FEMALE RATS RESPECTIVELY AFTER 90 DAYS. The histological features of the different organs were displayed in the figure for the male and the female rats, respectively. SMD 500 mg/kg Liver showing mild vascular degeneration with nucleus condensed. Kidney, spleen, and heart showed no observable histological changes in male as well as female groups. SMD 500 mg/kg Lung showed accumulation of foamy macrophages and mild feathery degeneration of hepatocytes. SMD 250 mg/kg and 100 mg/kg were showed no observable histological changes in both the groups.
Assessment of Chronic Toxicity of Phytoconstituent in Male and Female Rats:
TABLE 14: EFFECT OF ORAL 180 DAYS OF REPEATED ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON BODY WEIGHT
Group | 0 days | 30 days | 60 days | 90 days | 120 days | 150 days | 180 days | |
Body weight in gm | ||||||||
Male | Male
control group |
190±1.18 | 192±0.16 | 197±1.62 | 201±0.60 | 204±0.55 | 209±0.53 | 211±1.21 |
SMD
250 mg/kg |
191±1.22*** | 189±0.70*** | 185±0.11** | 187±0.57** | 186±0.49** | 181±0.31** | 179±0.35** | |
SMD
100 mg/kg |
190±1.75*** | 194.5±1.87*** | 192±0.27*** | 190±0.43*** | 189±0.59*** | 189.5±0.53*** | 191±0.11*** | |
SMD
50 mg/kg |
190±0.54*** | 189±1.21*** | 191±0.23*** | 194±0.59*** | 194.5±0.43*** | 196±0.32*** | 197±0.35*** | |
Female | Female
control group |
190±0.60 | 200±1.17 | 201±1.76 | 201±0.60 | 202±0.55 | 207±0.42 | 209±1.19 |
SMD
250 mg/kg |
192±0.58### | 191±0.21### | 189±0.89## | 187±0.50## | 187.5±0.84## | 185.5±0.31## | 182±1.99## | |
SMD
100 mg/kg |
189±0.24### | 192±0.34### | 192±0.21### | 190±0.55### | 189±0.99### | 191.5±0.25### | 190±1.25### | |
SMD
50 mg/kg |
193±0.17### | 194±0.44### | 196±0.14### | 199±0.64### | 201±1.20### | 205±0.37### | 206.5±0.44### |
Values are expressed as Mean ± S.E.M (n=20). ***P<0.05 & **P<0.05 when compared with repeated administration of SMD Male rats group concerning the male control group, ###P<0.05 & ##P<0.05 when compared with repeated administration of SMD Female rats group concerning the female group. The body weights of repeated oral administration rats with dose of SMD were displayed on Table for the male and the female rats respectively, data didn’t show much variation between the groups with respective control group even after 180 days however there were some mild changes dose of SMD 250 mg/kg observed within the male and the female groups.
GRAPH 4: EFFECT OF ORAL ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON PLASMA GLUCOSE LEVELS AND LIPID PROFILES. Values are expressed as Mean ± S.E.M (n=20). Examination of plasma glucose and lipid profiles; TG, TC, HDL, LDL, and VLDL did not show much variation (P<0.05) between the treated groups even after 180 days and their respective control groups. But both the group were showed significant variation (P<0.05) between TG, TC, HDL, LDL and VLDL concentrations in the treated groups when we were compared those in the normal control group.
TABLE 15: EFFECT OF ORAL ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON SERUM TOTAL PROTEIN, ALBUMIN AND GLOBULIN
Group | Total Protein (TP) gm/dl | Albumin gm/dl | Globulin gm/dl | |
Male | Male control group | 5.5±0.55 | 3±0.04 | 2.5±0.25 |
SMD 250 mg/kg | 6.5±0.22** | 4.1±0.06*** | 5.6±0.42** | |
SMD 100 mg/kg | 5.6±0.21*** | 3.3±0.04*** | 4.7±0.41** | |
SMD 50 mg/kg | 5.3±0.01*** | 2.9±0.01*** | 2.7±0.36*** | |
Female | Female control group | 5.4±0.14 | 3.1±0.03 | 2.2±0.77 |
SMD 250 mg/kg | 8.2±0.24## | 4.3±0.08### | 5.4±0.22## | |
SMD 100 mg/kg | 7.6±0.23### | 3.8±0.01### | 4.2±0.44## | |
SMD 50 mg/kg | 5.7±0.78### | 3.2±1.20### | 2.9±1.29### |
Values are expressed as Mean ± S.E.M (n=20) ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 180 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 180 days. Total protein and globulin were got mild changes in treated groups’ dose of SMD 250 mg/kg P<0.05. Rest parameters showed normal with their respective groups.
TABLE 16: EFFECT OF ORAL ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON SERUM TOTAL BILIRUBIN, DIRECT BILIRUBIN, BLOOD UREA, CREATININE, AND URIC ACID
Group | Bilirubin total mg/dl | Bilirubin direct mg/dl | Blood Urea mg/dl | Serum Creatinine mg/dl | Uric acid mg/dl | |
Male | Male control group | 0.4±0.53 | 0.3±0.02 | 28±0.21 | 0.7±0.11 | 2.6±0.02 |
SMD 250 mg/kg | 0.7±0.25** | 0.5±0.09*** | 35±0.86** | 0.5±0.21*** | 8.3±0.36** | |
SMD 100 mg/kg | 0.5±0.32*** | 0.3±0.01*** | 29±0.24*** | 0.3±1.11*** | 5.2±0.34** | |
SMD 50 mg/kg | 0.3±0.51*** | 0.2±0.10*** | 27±0.11*** | 0.3±0.11*** | 3.1±0.54*** | |
Female | Female control group | 0.4±0.22 | 0.2±0.01 | 30±0.21 | 0.5±0.01 | 2.7±0.45 |
SMD 250 mg/kg | 0.8±0.31## | 0.5±0.23### | 36±0.23## | 0.7±0.24### | 8.1±0.57## | |
SMD 100 mg/kg | 0.4±0.42### | 0.3±0.14### | 28±0.11### | 0.6±0.26### | 5.3±0.69## | |
SMD 50 mg/kg | 0.2±0.32### | 0.2±0.11### | 31±0.03### | 0.3±0.01### | 2.9±0.97### |
Values are expressed as Mean ± S.E.M (n=20). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 180 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 180 days. Blood urea and uric acid were significantly increased in treated groups dose of SMD 250mg/kg were observed P<0.05. Moreover, a mild increase in bilirubin was observed P<0.05 in the rats treated with SMD 250 mg/kg compared with their respective control group.
TABLE 17: EFFECT OF ORAL ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON SERUM SODIUM, POTASSIUM, CHLORIDE, AND ALKALINE PHOSPHATE
Group | Sodium (mmol/L) | Potassium (mmol/L) | Chloride (mmol/L) | Alkaline Phosphate (ALP) U/L | |
Male | Male control group | 141±2.42 | 4±0.46 | 102±0.14 | 158±0.21 |
SMD 250 mg/kg | 139±1.13*** | 3±0.18*** | 91±0.34*** | 171±0.22** | |
SMD 100 mg/kg | 142±0.68*** | 3.7±1.31*** | 98±1.14*** | 144±0.21*** | |
SMD 50 mg/kg | 140±0.86*** | 3.9±0.11*** | 99±1.22*** | 147±0.25*** | |
Female | Female control group | 140±0.25 | 3.9±0.33 | 103±1.26 | 156±0.78 |
SMD 250 mg/kg | 137±0.08### | 3.2±0.31### | 95±0.53### | 179±1.18## | |
SMD 100 mg/kg | 138±0.86### | 3.7±0.85### | 99±0.32### | 155±1.98### | |
SMD 50 mg/kg | 141±0.25### | 3.9±0.14### | 101±0.60### | 149±1.66### |
Values are expressed as Mean ± S.E.M (n=20). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 180 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 180 days. Examination of electrolytes; Sodium, Potassium, and Chloride didn’t show much variation between the treated groups P<0.05 and their respective control but there were mild changes P<0.05 in alkaline phosphate dose of SMD 250 mg/kg with their respective normal control group.
TABLE 18: EFFECT OF ORAL ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON HAEMATOLOGICAL PARAMETERS
Group | Hb (gm/dl) | WBC (c/cmm) | RBC (m/cmm) | Neutrophils (%) | Lymphocyte (%) | Platelet (lakh/cmm) | |
Male | Male
control group |
16±1.14 | 8.600±0.21 | 8.5±0.45 | 60±0.01 | 35±0.16 | 3.22±1.14 |
SMD
250 mg/kg |
14±0.21*** | 7.338±0.25** | 10.2±0.74** | 55±0.24*** | 22±0.19** | 2.22±1.24*** | |
SMD
100 mg/kg |
14.15±0.25*** | 8.000±1.27*** | 9.92±0.34** | 51±0.45*** | 29±0.14*** | 2.67±0.11*** | |
SMD
50 mg/kg |
15.9±1.18*** | 8.500±1.13*** | 9.47±0.77*** | 59±0.23*** | 30±0.85*** | 2.81±0.33*** | |
Female | Female
control group |
13.54±1.28 | 8.502±1.38 | 8.6±0.33 | 57±0.47 | 28±0.65 | 3.10±0.12 |
SMD
250 mg/kg |
14.2±0.23### | 7.312±1.28## | 10.47±0.41## | 51±0.21### | 24±0.35## | 2.14±0.55### | |
SMD
100 mg/kg |
14.5±0.14### | 8.544±0.57### | 8.2±0.82### | 53±0.33### | 25±0.14### | 2.64±0.65### | |
SMD
50 mg/kg |
14.9±0.17### | 8.589±0.14### | 8.5±0.84### | 54±0.93### | 23±0.24### | 2.58±0.97### |
Values are expressed as Mean ± S.E.M (n=20). ***P<0.05, **P<0.05 when compared with treated male rats group with their control group with after 90 days, ###P<0.05, ##P<0.05 when compared with treated female rats group with their control group after 90 days. Examination of hematological levels did not show much variation between the treated groups and their respective control groups.
Histological Examination of Haematoxylin and Eosin (H&E) Stained Tissues:
FIG. 4: EFFECT OF ORAL ADMINISTRATION OF SMD 250 mg/kg, 100 mg/kg AND 50 mg/kg ON THE DIFFERENT ORGANS OF MALE RATS AND THE FEMALE RATS RESPECTIVELY AFTER 180 DAYS. The histological features of the different organs were displayed in the figure for the male and the female rats, respectively. SMD 250 mg/kg Liver showed normal architecture, sinusoidal congestion, portal veins dilated and congested with focal portal inflammation. No necrosis. Spleen showed normal; Lung showed dilatation of alveoli. Kidney and Heart were showed no pathological changes in all the groups. SMD 100 mg/kg and 50mg/kg were showed the liver normal architecture, Spleen normal white pulp, and lung showed normal alveoli. Kidneys were showed normal glomeruli tubules, normal adrenal also saw. Heart showed normal endocardium, myocardium, no pathological changes in male and the female groups.
DISCUSSION: For centuries, natural products, such as medicinal plants have been the basis for the treatment of various ailments 9. In screening natural products for the pharmacological activity, assessment and evaluation of the toxic characteristics of a natural product extract, fraction, or compound are usually an initial step. Regardless of the pharmacological beneficial effects of Momordica dioica, detailed knowledge about the chronic toxicology of this famous herb is lacking.
Hence, the current study was undertaken to evaluate and focus on the acute, sub-acute, sub-chronic and chronic toxicity of saponin of M. dioica in an animal model. In this study saponin M. dioica extract at a dose of 5000 mg/kg had no adverse effect on the tested in rats up to 14 days of observation. Therefore, this study indicates that saponin M. dioica does not cause acute toxicity effects at the dose tested and with an LD50 value of 5000 mg/kg. However, many medicinal plants have also been reported to be toxic to both humans and animals. Therefore, it should be emphasized that the traditional use of any plant for medicinal purposes, by no means, guarantees the safety of such plant. Furthermore, the data of the sub-acute, subchronic and chronic toxicity studies on medicinal plants or preparations derived from them should be obtained to increase the confidence in its safety to humans, particularly for use in the development of pharmaceuticals. Consequently, in this study, the sub-acute toxicity saponin M. dioica was evaluated in male and female rats at doses of 1000 mg/kg, 500 mg/kg and 250 mg/kg for 28 days.
Administration of extracts of Saponin M. dioica for 28 days produced no clinical sign for toxicity in rats with dose of 500 mg/kg, and 250 mg/kg of Saponin M. dioica and data didn’t show much variation between their respective control groups even after 28 days but there were some mild changes observed at dose of Saponin M. dioica 1000 mg/kg. Further confirmation by sub-chronic toxicity saponin M. dioica was evaluated in male and female rat doses of 500 mg/kg, 250 mg/kg and 100 mg/kg for 90 days. Administration of oral extracts of Saponin M. dioica for 90 days produced no clinical signs for toxicity or mortality. Also, the treated rats didn’t show any significant alternation, but there were some significant changes observed at a repeated dose of 500 mg/kg with their respective control groups after 90 days. This finding further confirmation by the administration of extract of saponin M. dioica 250 mg/kg, 100 mg/kg and 50 mg/kg for 180 days produced no clinical signs for toxicity or mortality. Therefore, the results recorded in this study demonstrated that the saponin M. dioica was safe, no toxic sign reported in doses of 250, 100 and 50 mg/kg.
CONCLUSION: In light of these findings, we may conclude that saponin of Momordica dioica is not toxic the doses of 250, 100 and 50 mg/kg/b.w. herein and did not produce any toxic signs or evident symptoms. Saponin of Momordica dioica did not cause any lethality or produce any remarkable histopathological signs or serum chemical alteration the doses mentioned.
The preliminary results suggest promising alternatives for exploring therapeutic and pharmaceutical interest in saponin of Momordica dioica with a reduction of possible adverse effects. Further studies to determine the effects of saponin of Momordica dioica extract on an animal foetus, on pregnant animals and their reproductive capacity are needed to complete the safety profile of this herb.
ACKNOWLEDGEMENT: I would like to express heart full thanks to my beloved Parents, Family, the guide for their support and blessings.
CONFLICT OF INTEREST: None
REFERENCES:
- Dias FDL and Takahashi CS: Cytogenetic evaluation of aqueous extracts of the medicinal plants Alpinia mutants rose (Zingerberaceae) and Pogostemum hyneanus benth (Labitae) on Wistar rats and Allium cepa (Liliaceae) root tip cells. Brazilian Journal of Genetics 1994; 17(2): 175-80.
- Jha DK, Koneri R and Samaddar S: Potential Bio-Resources of Momordica dioica Roxb: A Review. Int J Pharm Sci Rev Res 2017; 45(2): 203-09.
- Ilango K, Maharajan G and Narasimhan S: Preliminary phytochemical screening and antibacterial activity of fruit pulp of Momordica dioica Roxb. (Cucurbitaceae) African Journal of Basic & Applied Sciences 2012; 4(1): 12-15.
- Salvi J and Katewa SS: Nutritional Composition of Momordica dioica fruits: As a wild vegeTable. J Food Pharm Sci 2015; 18-23.
- Sharada LD and Somshekhar SK: Isolation and characterization of phytoconstituents from Chlorophytum borivilianum. Pharmacognosy Res 2010; 2: 343.
- Gini TG and Jeya-Jothi G: Preliminary phytochemical screening. Int J of Pharmacognosy and Phytochemical Res 2013; 5(3): 200-14.
- Anjamma M and Bhavani LN: Comparative phytochemical constituents evaluation from the fruit extracts of Momordica charantia L. and Momordica dioica Roxb. Int J Curr Biotechnology 2015; 3(8): 17-21.
- Burke RW, Diamondstone BI, Velapoldi RA and Menis O: Mechanisms of the Liebermann-Burchard and Zak Color Reactions for Cholesterol. Clin Chem 1974; 20(7): 794-01.
- Ridtitid W, Sae-Wong C, Reanmongkol W and Wongnawa M: Antinociceptive activity of the methanolic extract of Kaempferia galanga Linn. in experimental animals. Journal of Ethnopharmacology 2008; 118(2): 225-30.
How to cite this article:
Jha DK, Koneri R and Samaddar S: Toxicity studies of a saponin isolated from the fruits of Momordica dioica in rats. Int J Pharm Sci & Res 2019; 10(10): 4462-76. doi: 10.13040/IJPSR.0975-8232.10(10).4462-76.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
Article Information
6
4462-4476
1306
1637
English
IJPSR
D. K. Jha *, R. Koneri and S. Samaddar
Department of Pharmacology, Karnataka College of Pharmacy, Bangalore, Karnataka, India.
deepakjha736@gmail.com
15 January 2019
10 April 2019
21 April 2019
10.13040/IJPSR.0975-8232.10(10).4462-76
01 October 2019