INTRODUCTION:

Diabetes mellitus is a multifaceted, persistent illness involving uninterrupted health care with multifactorial risk-reduction approaches beyond glycemic control. Continuing patient personal execution education and support are serious to preventing acute impediment and falling the risk of enduring complications.

Considerable facts exist that chains a range of interference to recover diabetes outcomes¹. Diabetes mellitus has developed to the third most “killer” chasing after cardio-/cerebro-vascular diseases and cancer². It is predictable that 5% of every fatality in the world is caused by diabetes, a digit which will be elevated by 50% in the next 10 years³. At the instance of systematic discovery, about 70% of the host’s total β-cell mass is damaged as an outcome of immune-mediated processes⁴. At present there is no convinced cure for diabetes, an overarching aim in the management of all varieties of diabetes is safeguarding and even promising regeneration of β-cells. Natural products isolated beginning from medicinal plant assets have been in use for the prevention and control of a variety of diseases/ pathologies, diabetes mellitus, cancers, heart disease and also high blood pressure⁵. Almost more than 800 species have been examined and their hypoglycemic effects reported. A number of conventional plants and herbs recognized to contain therapeutic efficiency for purpose of many diseases and disorders. Moringa oleifera (Moringaceae) an extremely familiar medicinal plant also well-known as Drumstick. Its seeds exhibited analgesic action, Antipyretic action⁶, ant-obesity activity⁷ and anthelmintic activity⁸. Its leaves proved antimicrobial activity⁹, Wound healing activity¹⁰, Analgesic activity^10-11, Antiulcer activity¹³, Hypotensive¹⁴, Diuretic activity¹⁵. Roots confirmed antifertility activity¹⁶, Anti-inflammatory result by Carrageenan induced rat paw edema method was shown by Extracts of stem bark of M. oleifera and also were revealed as cotton pellet granuloma creation analgesic efficacy¹⁷. M. oleifera stem bark Extracts were reported antioxidant and Cytotoxic potential¹⁸. Diversity of disorders was generated through the release of lysosomal enzymes in inflammation. Acute or chronic inflammation is linked to the extracellular action of these enzymes. A parameter of anti-inflammatory activity of the drug is therefore is taken as lysis of human red blood cell membrane¹⁹. An extensive variety of medicinal compounds like flavonoids rutin, butadion and escinol have membrane stabilizing^20-22. A number of researchers have been employed Erythrocytes as a representation method for the study of interaction of drugs with membranes. Membrane stabilizing property was shown by diverse classes of drugs shown like anesthetics, NSAIDs. A practical In vitro model for evaluating the anti-inflammatory activity of compoundsis Stabilization of HRBC membrane by drugs against hypotonicity stimulated haemolysis^23-25. Antimicrobial activity was also shown by M. oleifera against selected microorganisms compared with standard as Gentamicin ²⁶. The aim of current research work is to find out the blood glucose level changes in normal and alloxan monohydrate-induced diabetic rats by extracts of M. oleifera. For these examinations, a recognized model, i.e., the alloxan induced diabetic rat, was used.

MATERIAL AND METHODS:

The research work was carried out in SMBT College of Pharmacy, Dhamangaon, Igatpuri, Nashik, 422403, India. The approval for study was obtained from the Institutional Animal Ethical Committee. CPCSEA Reg. No. 1329/PO/Re/S/10/CPCSEA wef 27/10/2016.

Plant Material:

The collection of M. oleifera (Moringaceae) stem bark was done from habitat area of Nashik, India. Dr. Diwakar P.G. Director, Botanical survey of India, Pune has done the recognition and authentication of the collected plant material. (Reference No. BSI/WC/Tech/ 2009/370).

Preparation of extract:

Stem bark was made dirt free dried out under shadow and powdered in grinder. Soxhlet apparatus was employed for sequential extraction of the powder (450g) first with petroleum ether, followed by chloroform and methanol by considering order of polarity. The yields were recorded as 0.89%, 3.6%, and 16.63% for the petroleum ether, chloroform and methanol respectively. The extracts have shown presence of sterols, glycosides, alkaloids, flavonoids, triterpenoids and tannins in introductory Phytochemical analysis.

Experimental design and Collection of samples:

For research work the Wistar strain of Albino rats (150-200 g) of both sexes were employed and were acquired from Haffkine Institute, Mumbai. Standard polypropylene cages were used for housing. Room temperature (24 ± 2 ºC; relative humidity 60%--70%) in a 12 h light-dark cycle was maintained. Standard laboratory food of Pranav agro Pvt. Ltd was used for the animals and water ad libitum. Withdrawal of Food was done before 12 h and through the experimental hours. The approval for study was obtained from the Institutional Animal Ethical Committee. Wistar albino male rats (150–200g) were used. Rats were provided the standard diet. Randomized the groups and followed by commencement of experiment. The rats were adapted for a period of one week under standard conditions of relative humidity, temperature, as well as dark/light cycle. Animals depicted as fasting were deprived of food for 16 h and water ad libitum. Retro-orbital plexus puncture method was employed for Blood samples collection. An electronic glucometer was used for blood glucose levels^27-30.

All the animals were haphazardly divided into the six groups with six animals in each group and treatment given as below

Group 1: Normal saline

Group 2: Diabetic (control) Vehicle

Group 3: Standard drug (glibenclamide, 10mg/kg per day p.o)

Group 4: 100mg/kg per day p.o methanolic extract M.orifrea

Group 5: 200mg/kg per day p.o methanolic extract M.orifrea

Group 6: 400mg/kg per day p.o methanolic extract M.orifrea

A single dose of alloxan monohydrate (150mg/kg) was given to the rats by i.p. route for inducing diabetes mellitus. Alloxan monohydrate dissolved in normal saline just earlier to injection. After 48 h, rats having plasma glucose levels more than 140mg/dl were incorporated in the reading. After 48 h of alloxan injection treatment was given as above. Blood sample were drained at weekly intermissions till the conclusion of study (i.e. 3 weeks). Fasting blood glucose assessment and body weight measured on 1, 7, and 21 days of the study. Fasting blood sugar level was estimated on 21^st day by cardiac puncture using ether as mild anesthetic agent from 12 h fasted rats³¹.

Statistical analysis:

The results of every the set of observations have been specified in terms of mean ± SEM. Analysis of variance (ANOVA) was used for determination of variation between the groups. Statistics was used with Dunnett’s test multiple comparisons test by means of Graph Pad in Stat version 5.00. The level of significance was set at P < 0.05.

RESULTS:

The insulin-secreting beta cells of pancreas were destructed by Alloxan and causes diabetes. Alloxan is particularly lethal to beta cells of pancreas shown In vitro studies, most important to the initiation of cell necrosis. Alloxan (150mg/kg, i.p.) administration tends to increase in fasting blood glucose levels, that holds for 21 Days. The observations were conducted for three weeks. For Group 1 levels remains as normal. For Group 2 sugar levels sustained as from 296mg/dl to 295mg/dl. For Group 3 drop in blood glucose levels from 295 mg/dl to 149mg/dl. For Group 4 drop in blood glucose levels from 294mg/dl to 237mg/dl. For group 5 drop in blood glucose levels from 295mg/dl to 232mg/dl. For group 6 drop in blood sugar levels from 290mg/dl to 229 mg/dl in 21 days for methanolic extract of M. oleifera. (Table1) Effect appears to attain maximum after two weeks of treatment and remains stable in third week. Vehicle control animals were shown to be robust in their body weight except diabetic rats showed prominent decrease in body weight during 21 days. Alloxan caused decrease in body weight.

Figure 1: The effect of M. Oleifera extracts on normal and alloxanized diabetic rats

DISCUSSION:

It is recognized that in the pathogenesis of diabetes mellitus one of the key associations of activation of free radical oxidation. A discrepancy takes place between prooxidants and antioxidants, leading to surplus of free radicals and the formation of products of free radical oxidation. It has been exposed that modify people’s diet, raise physical activity and direct to the loss of surplus body weight can avoid type 2 diabetes. For instance, the Diabetes Prevention Program in the USA³², the Finnish Diabetes Prevention Study³³and the Chinese Da Qing Study established that vigorous intervention, lasting 2 to 6 years, might have extended help for glycemic and cardiovascular results as compare with previous 10 to 20 years³⁴. Diabetes is an exigent disease; its physical condition harm on human is escalating worldwide. The call for medication of diabetes is gradually tracked by professionals. Synthetic drugs such as metformin and glibenclamide are having lot of adverse effects. This recommends for herbal remedy with no or nominal side effect to manage and treat diabetes³⁵. Type 2 diabetes can be originated by different tissues also comprises both genetic and environmental issues that affect β-cell role and tissue of pancreas, muscles as well as liver³⁶. Administration of natural products with oral hypoglycemic drugs for the control of diabetes may create a potential drug herb interaction that may have helpful or adverse effects. It is usually supposed that the use of herbs with medicine makes improved effect and decreases the adverse effects of the drugs³⁷. Phytochemicals symbolize a natural source of compounds from which novel and potent drugs could be intended for diabetes treatment. This is predominantly significant taking into consideration that metformin, the first line drug used to manage type 2 diabetes, has objectionable gastrointestinal side effects together with diarrhea, flatulence and abdominal discomfort³⁸. In spite of the hopeful biological effects of abundant hypoglycemic phytochemicals explained, the low absorption and bioavailability of these materials remains an obstruction for their restraint as prospect antidiabetic agents in the treatment of diabetes. Numerous approaches i.e. absorption enhancer, self-microemulsion, and solid lipid nanoparticles, were employed to augment the constancy and bioavailability of phytochemicals³⁹. The treatment of diabetes with drugs of plant origin that confirmed much safer than synthetic drugs is an essential part of many traditions throughout the world and grown value in recent time. India has an affluent history of using a variety of potent herbs and herbal materials for treating various diseases including diabetes⁴⁰.

After alloxan injection of the rats gave confirmation of considerable raise in blood glucose levels shown in the present study. Etuk, Muhammed and Adeyi established raise in glucose levels to the hasty oxygen species provoked by alloxan; this, in amalgamation with a concurrent enormous elevation in cytosolic calcium concentrations tends to rapid injury of pancreatic islet cells plus linked reduction in synthesis/release of insulin^41-42. Numerous chemically modified drugs have been developed for the management of diabetes. Nevertheless, these drugs contain restrictions in terms of efficacy and side effects. Consequently, there is a great deal attention in finding out natural treatments devoid of negative side effects in diabetic patients. Natural products appear to be the solution⁴³. Flavonoids are one of the most abundant and common group of phenolics in higher plants^44-46. A number of them, due to their phenolic structure, are recognized to be concerned in the remedial progression of free radical-mediated diseases including diabetes^47-48.

The use of flavonoids, diminishes the risk of diabetes. Also, Saponins have the capacity to decrease elevated plasma blood glucose, thus, making it supportive treatment in diabetes mellitus. The hypoglycemic effect of saponin is through restoration of insulin response, enhancement in insulin shows, increase plasma insulin and induction of insulin release from the pancreas⁴⁹. In the similar way, the existence of Triterpenes restrains enzymes involved in glucose metabolism and avoids the increase of insulin resistance therefore regularizing insulin levels⁵⁰. As per previous phytochemical work authenticates the presence of flavonoids, triterpenoid saponin and other phytoconstituents present in M. olifera bark²⁰. So due to this phytoconstituents and antioxidant activity^18,51 of M. olifera shows hypoglycemic effect on normal and alloxanized diabetic rats. Future work is to isolate phytoconstituents which responsible for hypoglycemic activity.

CONCLUSION:

From the current study it was attempted to confirm that methanolic extract of M. olifera bark is competent enough to demonstrate the decrease in blood glucose level. This study was confirmed in normal and alloxan induced diabetic rats. The mechanism of action could not be presented in detail because it is very difficult and also study was not designed in that view.

ACKNOWLEDGEMENT:

Authors are also thankful Principal, SMBT College of Pharmacy, Dhamangaon, India for providing necessary facilities.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 07.01.2021 Modified on 15.02.2021

Res. J. Pharma. Dosage Forms and Tech.2021; 13(2):95-99.

DOI: 10.52711/0975-4377.2021.00017

Gr. No.	Treatment	Fasting blood glucose level (mg/dl)
Gr. No.	Treatment	0	7	14	21
1	Normal	90.167±5.36	92.000±4.31	91.333±4.51	93.833±2.87
2	Diabetic Control (Alloxan+Vehicle)	296.167±8.67	291.667±4.20	293.000±6.54	295.167±11.20
3	Alloxan + glibenclamide	295.333±11.23	211.500±12.60***	158.167±10.29***	149.833±9.96***
4	Alloxan +MO100	296.833±12.14	245.333±18.74*	235.333±10.01***	227.500±11.47***
5	Alloxan +MO200	294.833±11.78	250.333±12.75*	229.000±10.87***	222.167±8.91***
6	Alloxan +MO400	300.000±6.67	244.167±13.25**	226.833±11.63***	219.500±10.47***