Pharmacological Evaluation of Leaf Extract of Terminalia bellerica with Moringa oleifera for its Synergistic Action on Anti-diabetic Activity and Anti-inflammatory Activity in Rats
Sk. Karishma*, K. Lakshmi, D. Eswar Tony, A. Narendra Babu, Rama Rao Nadendla
Department of Pharmacology, Chalapathi Institute of Pharmaceutical Sciences, Guntur. Andhra Pradesh
Aim and Objective: To evaluate the pharmacological action of leaf extract of Terminalia bellerica with Moringa oleifera for its synergistic action on Anti Diabetic activity and Anti-inflammatory activity in rats. Methodology: The Pharmacological evaluation of standardized aqueous extract of the leaves of Terminalia bellerica with Moringa oleifera was carried by using the following Stimuli models: a)Antidiabetic activity by Streptozotocin Induced Diabetes, b) Anti-inflammatory activity by Egg Albumin Induced Paw Edema. Results: The parameters of various activities to assess anti diabetic and anti inflammatory activities were evaluated and found That, The combination of extracts possess a better response then alone. Conclusion: The aqueous leaf extract of the combination of Terminalia bellerica with Moringa oleifera has shown significant anti diabetic and anti-inflammatory activities when compared with standard and extract alone.
Diabetes mellitus is a group of metabolic alterations characterized by hyperglycemia resulting from defects in insulin secretion, action or both1. Diabetes is a chronic disease without a cure and it is associated with significant morbidity and mortality. Though there are various approaches to reduce the ill effects of diabetes and its secondary complications, herbal formulations are preferred due to lesser side effects and low cost. Many allopathic medicines are available for the treatment of diabetes, includes sulphonylureas, biguanides, meglitinides and thiazolidinediones but some how they have their own adverse effects like hypoglycemia, nausea, vomiting, hyponatremia, flatulence, diarrhea or constipation, alcohol flush, headache, weight gain, lactic acidosis, pernicious anemia, dyspepsia, dizziness and joint pain. So instead of allopathic drugs, herbal drugs are the best choice which is having fewer side effects and non toxic properties.2
Inflammation is the response to injury of cells and body tissues through different factors such as infections, chemicals, thermal and mechanical injuries.3 It can be classified as either acute or chronic, and involves a cascade of biochemical events comprising the local vascular system, the immune system, and different cell types found in the injured tissue. Acute inflammation is the initial response and is characterized by the increased movement of plasma and innate immune system cells, such as neutrophils and macrophages, from the blood into the injured tissues.4 Chronic inflammation concerns a progressive change in the type of cells present at the site of the inflammatory reaction and is characterized by simultaneous destruction and healing of the injured tissue.
Terminalia bellerica also referred to as, Beleric Myrobalan belonging to family Combretaceae. Terminalia bellerica is used in traditional medicine due to the wide spectrum of pharmacological activities associated with the biologically active chemicals present in this plant. The phytoconstituents isolated from various parts of the plant include alkaloid, coumarin, flavones, steroids (β- Sitosterol), lignans (termilignan, thannilignan), tannins (gallic acid, ellagic acid), glycosides (fructose, sucrose, galactose), terpenoid (belleric acid and chebulagic acid), saponin (bellericoside and bellericanin). Terminalia bellerica is one such plant showing multifarious medicinal properties viz. analgesic activity, antibiofilm activity, anticancer activity, antidepressant activity, antidiabetic activity, antidiarrhoeal activity, antiulcer activity, immunomodulatory activity, antispasmodic and bronchodialatory activity, antifertility activity, antihypertensive activity, antifungal, antimicrobial activity, anti-inflammatory activity, antioxidant activity.5
Moringa oleifera Lam, popularly called “the miracle tree”, is a monogeneric plant of the family Moringaceae. Moringa oleifera is rich in minerals, carbohydrates, proteins, fats, moisture, crude fiber, and ash contents M. oleifera leaves have been used by local traditional healer in treatment of various ailments such as diabetes, gastric discomfort, stomach ulcer, diarrhea, dysentery and skin infections. The leaves have also been found to possess antitumor, antipyretic, antiepileptic, anti-inflammatory, anti-ulcer, anti-hypertensive and anti-oxidant properties. Phytochemical analysis of Moringa oleifera showed the presence of flavonoid, anthraquinone, alkaloids, saponins, steroids, terpenoids, cardiac glycoside, anthocyanintannins and carotenoid in aqueous extract.6
MATERIALS AND METHODS:
Collection of leaves:
The leaves of Terminalia bellerica and Moringa oleifera Lam, were collected from Chalapathi Institute of Pharmaceutical Sciences and authenticated by DR. P. Satyanarayana Raju, Dept. of botany and microbiology, Acharya Nagarjuna University, Guntur.
Preparation of plant extract:
Fresh leaves of Terminalia bellerica and Moringa oleifera Lam, were washed and shade dried at room temperature for 10-15 days and powdered with the help of a mechanical grinder and 30gm of leaves powder was extracted separately with 250ml of distilled water to obtain aqueous extract with the help of Soxhlet apparatus. The extract was collected in Petri dishes and evaporated till dry at 40ºC in an incubator. Then the extract was sealed with aluminium foil and stored at 4ºC for further experimental work.
Sprague-dawely rats are weighing about 160g were obtained from animal house, Chalapathi Institute of Pharmaceutical Sciences, Guntur. The experimental protocol was approved by the Institutional Animal Ethics Committee. The animals were housed in large spacious cages and were fed with standard pellet diet and water ad libitum. The animals were maintained under their respective controlled temperature conditions for 30 days before experiment. The animals were allowed to acclimatize to laboratory conditions for 48 h before the start of the experiment.
Pharmacological evaluation of leaf extract of Terminalia bellerica with Moringa oleifera for its synergistic action on anti diabetic and anti inflammatory activities in rats has been approved (approval No: 09/IAEC/CIPS/2017-18; Dt: 29/11/2017) by the institutional animal ethics committee (IAEC),(Reg.No:1048/PO/Re/S/07/CPCSEA) Chalapathi Institute of Pharmaceutical Sciences, Guntur.
Screening for Anti Diabetic Activity:
Sprague-dawely rats either both sex were divided into five groups containing five animals in each.
· Group-1-Control Group (0.9% Normal saline 5ml/kg orally)
· Group-2-Inducing group (60mg/kg of STZ i.p)
· Group-3–Aqueous leaf extract of Terminalia bellerica (100mg/kg orally)
· Group-4–Aqueous leaf extract of Terminalia bellerica and Moringa oleifera (100mg/kg orally)
· Group-5-Standard group (500µg/kg of Glibenclamide orally)
Streptozotocin Induced Diabetes:
Diabetes was induced by single intraperitoneal injection of a freshly prepared solution of STZ at a dose of 60 mg/kg bodyweight.7 Diabetes was developed and stabilized in STZ treated rats over a period of 7days. Blood sample was collected from the tail vein and blood glucose levels were determined by using an Accu-Chek Active glucometer at 0hr, 1h, 2h, 4h and 6h, after STZ injection. The rats with blood glucose level above 250 mg/dl were considered to be diabetic and were used in the experiment.8
Screening for Anti Inflammatory activity:
Sprague-dawely rats either both sex were divided into five groups containing five animals in each.
· Group-1- Control group (0.9% Normal saline 5ml/kg orally)
· Group-2-Inducing group (0.1 ml of 1% fresh egg albumin)
· Group-3-Aqueous leaf extract of Terminalia bellerica (100mg/kg orally)
· Group-4-Aqueous leaf extract of Terminalia bellerica and Moringa oleifera(100mg/kg orally)
· Group-5-Standard group(Indomethacin 20mg/kg i.p)
Egg albumin-induced paw edema in rats:
The acute inflammation of the hind paw was induced in each of the rats by injecting 0.1ml /kg body weight of fresh egg albumin into the sub plantar surface of the right hind paw.9Change in the paw volume was recorded using a digital plethysmometer–PLM01, at 0min, 30min, 60min, 120min.10
The IITC 520 Plethysmometer was used to measure the paw volume. It consists of a water cell which is fitted on to a plexi glass stand. This water cell has an inlet in which the paw is dipped and an outlet with a stopcock. The water cell is connected to an electronic display which shows the volume of displacement of water in millilitre (ml) from the water cell when paw is dipped into water cell which is taken as paw volume. The paw is inserted into water, contained in a special water cell of which the resistance is changed due to the immersion of the animal’s paw. This resistance change is calibrated in ml and displayed on the read out in ml with a resolution of 0.1 ml.
Fig 1: Mean Observation of anti-diabetic activity of Control group
Fig 2: Mean observation of Anti Diabetic activity of inducing group
Fig 3: Mean observation of anti Diabetic activity of Test-I
Fig 4: Mean observation of Anti Diabetic activity of Test-II
Fig 5: Mean observation of Anti Diabetic activity of Standard group
Fig 6: Mean observation of Anti Inflammatory activity
In case of both anti-diabetic and anti-inflammatory, the test I and II, when compared with standard, showed a very good potency of having the activities by decreasing the severity of inflammation and also a marked reduction in glucose concentrations in blood. This is therefore confirmed that the combination of extracts have a better effect than the extract given alone.
The aqueous Leaf extract of combination of Terminalia bellerica with Moringa oleifera has shown significant Anti Diabetic and Anti-inflammatory activity when compared with standard and extract alone.
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Received on 28.11.2018 Modified on 27.12.2018
Accepted on 14.01.2019 © RJPT All right reserved