B-3337 [1-8] Indian J. Anim. Res., Print ISSN:0367-6722 / Online ISSN:0976-0555
AGRICULTURAL RESEARCH COMMUNICATION CENTRE
www.arccjournals.com/www.ijaronline.in
Protective effect of Psidium guajava leaf ethanolic extract against streptozotocin-induced diabetes and lipidosis in rats R. Manikandan1*, A.Vijaya Anand2, P. Sampathkumar3 and N. Manoharan4 Department of Biochemistry, M.I.E.T Arts and Science College, Trichy- 620 007, Tamilnadu, India. Received: 14-11-2016 Accepted: 12-05-2017
DOI: 10.18805/ijar.B-3337
ABSTRACT This study was conducted to find out the anti-diabetic and hypolipidemic potential of ethanolic extract of Psidium guajava Linn leaves and its one of the important compound of caryophyllene in a streptozotocin (STZ) induced diabetic rats. The rats were divided into eight groups. Diabetes was induced by STZ at a dosage of 60 mg/kg b.w. The various dosages of extract (100, 200, 300 mg/kg b.w), caryophyllene (300 mg/kg b.w) were injected and glibenclamide (3 mg/kg b.w) is used as a standard drug. After the treatment of the extract and caryophyllene the levels of blood glucose, HbA1c, glucose-6-phosphatase, fructose-1,6-bis phosphatase, total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), high density lipoprotein (HDL) were determined. Pancreatic tissue changes were noticed in all the groups. The present study showed that there was a significant reduction in blood glucose, HbA1c, glucose-6-phosphatase, fructose-1,6-bis phosphatase, TC, TG, LDL, VLDL and the enhancement was noticed in the level of insulin, glucokinase and HDL. Pancreas was damaged in diabetic induced rats and it altered to normal size and shape in the plant extract treated and caryophyllene treated groups. The results proved that the STZ induced diabetic rat models had revealed anti-diabetic and hypolipidemic effect treated with ethanolic leaf extract of P. guajava and caryophyllene. Key words: Antihyperglcemic, Caryophyllene, Hypolipidemic, Psidium guajava, Streptozotocin. . subtropical areas of the world and it adapts to all climatic INTRODUCTION conditions (Stone, 1970). P. guajava has mainly used in the Diabetes is a metabolic disorder resulting from the treatments of diarrhoea and tooth ache in an ancient times, defects in insulin secretion, insulin action, or both, which but researchers are surprised that the pharmacological increases the concentration of blood glucose (Rajiv Gandhi activities and it exhibits of anticough, antispasmodic, and Sasikumar 2012). In worldwide there are 347 million hepatoprotection, cancer prevention, cardioprotective and people are affected by diabetes (Danaei et al., 2011). The anti-inflammatory effects (Gutierrez et al., 2008). elevation of glucose level causes the oxidative stress, it leads Mexico and other Central American countries such to the oxidation, glycation and dyslipidemia (El-Hilaly et al., as Caribbean, Africa and Asia used the P. guajava for various 2006). Dyslipidemia in diabetic patients is more prevalent purposes from a long time in history for its medicinal values. and subsequently develop cardiovascular disease (Saravanan The leaves decoction of P. guajava is highly used in Colombia et al., 2003). Nearly 50% of people with diabetes die of and Mexico for curing the diarrhoea, dysentry, wounds, ulcers cardiovascular disease (Morrish et al., 2001). A number of etc (Heinrich et al., 1998). In Latin America and Mozambique oral hypoglycemic agents are available, but it causes the lot were used the leaves decoction for the treatment of diarrhoea of side-effects. Nowadays, the herbal products symbolize and stomach ache (Pondikis, 1996). The South African safety in contrast to the synthetic drugs. In developed peoples used the leaves decoction to control the diabetes countries, 25% of total drugs which are plant drugs, while in mellitus and hypertension (Oh et al., 2005). In USA the leaf developing countries, nearly 80% of plant constituents are decoction were used for its antibiotic nature (Smith and Nigel, used in total drugs (Joy et al., 1998). In the rest of the country, 1998). The decoction and the paste of the whole plant is used the India has the more economical importance of medicinal as a medicine for skin problems, painful menstruation, plants. premature labour in women, wounds etc in Tahiti and Sanoa P. guajava Linn. is an important plant in the (World Health Organization 1998). In Brazil the mashed Myrtaeceae family. P.guajava grows in all the humid and decoction of fruit, flower, and leaves were used for cholera, *Corresponding author’s e-mail:
[email protected] 1 Department of BioChemistry, M.I.E.T Arts and Science College, Trichy- 620 007, Tamilnadu, India. 2 Department of Human Genetics and Molecular Biology, Bharathiar University, Coimbatore-, Tamilnadu, India 3 Depatrment of Chemistry and Biosciences, SASTRA University, Kumbakonam-612001.TamilNadu, India. 4 Department of Marine Science, Bharathidasan University, Trichy-620 024. Tamil nadu, India.
2
INDIAN JOURNAL OF ANIMAL RESEARCH
gastric insufficiency, skin problems, vaginal discharge etc (Holetz et al., 2002). In India, the decoction of leaves were used to cure rheumatism (Hernandez 1971).
Standards and Technology Mass Spectral database (NISTMS). The percentage of each component was calculated from relative peak area of each component in the chromatogram.
Leaves of this plant are reported to contain caryophyllene, á-pinene, â-pinene, â-bisabolene, quercetin, cineol, caryophyllene oxide, limonene, menthol, â-copanene (Zakaria and Mohd 1994). Therefore the present study was investigated the anti-diabetic activity and hypolipidemic activity of ethanolic extract of P. guajava leaves in STZ induced diabetic rats.
Animals: Male albino rats of 6-8 weeks age, weighing 150180g, were used. The animals were kept in clean, dry plastic cages and fed with standard pellet diet and water. This study was carried out in the animal house of Srimad Andavan Arts and Science College, Trichy (CPCSEA approval No-790/ 03/ac/CPCSEA) and this study was approved by the Institutional Ethical Committee. The animals were divided into eight groups with six rats each. Commercially available caryophyllene is used for the treatment of group VIII.
Caryophyllene is a natural volatile sesqueterpene with a molecular formula of C15H24. It is mainly present in various plant species such as Syzygium aromaticum, Piper nigram, Cannabis sativa, Cinnamon zeylanicum, Ocinummic ranthum and Origanum vulgare. It is present in many spices used by the Indian and Chinese peoples. It has various pharmacological effects such as anti-depressant (Bahi Amine et al., 2014), anti-inflammatory (Gertsch et al., 2008), neuroprotective (Guimaraes and Adriano, 2012), anti-viral, anti-oxidant and anti-cancer (Saad Sabbar Dahham et al., 2016). MATERIALS AND METHODS Plant material and extraction: The fresh P. guajava leaves were collected locally and authenticated by Botanist at Rapinat Herbarium, St. Joseph College, Trichy, Tamil Nadu, India. The shade dried P. guajava leaves were powdered mechanically and stored in an airtight container. The extraction of P. guajava leaves were done by a hot percolation method with Soxhlet apparatus. Ethanol was used as a solvent. About 100 gm of the powder of the plant materials was extracted with 600 ml of ethanol. The extract was concentrated to dryness under controlled temperature 4050°C. The percentage yield was found to be 10.15%. 30 g powdered sample of P. guajava leaves were soaked and dissolved in 75 ml of ethanol for 24 hours. Then the filtrates were collected by evaporating under liquid nitrogen. The GCMS analysis was carried out using a Clarus 500 Perkin-Elmer (Auto System XL) Gas Chromatograph equipped and coupled to a mass detector Turbo mass Gold-Perking Elmer Turbomas 5.2 spectrometer with an Elite-1 (100% Dimethyl ply siloxane), 300 m X 0.25 mm Xx 1 ìmdf capillary column. The instrument was set to an initial temperature of 110°C, and maintained at this temperature for 2 min. At the end of this period, the oven temperature was raised upto 280°C, at the rate of an increase of 5°C/min, and maintained for 9 min. The injection port temperature was ensured as 250°C and helium flow rate as 1 ml/minute. The ionization voltage was 70 eV. The samples were injected in split mode as 10:1. Mass Spectral scan rage was set at 45-450 (mhz). The chemical constituents were identified by GC-MS. The fragmentation patterns of mass spectra were compared with those stored in the spectrometer database using National Institute of
Group I : Normal rats (saline 2 ml/kg body weight). Group II : Diabetic control rats (STZ induced as 60 mg/ kg b.w.). Group III : Normal rats were fed with plant extract for 45 days (300 mg/kg b.w.). Group IV : Diabetic induced animals were fed with plant extract for 45days (100 mg/kg b.w.). Group V : Diabetic induced animals were fed with plant extract for 45days (200 mg/kg b.w.). Group VI : Diabetic induced animals were fed with plant extract for 45days (300 mg/kg b.w.). Group VII: Diabetic induced animals were fed with glibenclamide (3 mg/kg b.w.). Group VIII: Diabetic induced animals were fed with caryophyllenefor 45days (300 mg/kg b.w.). Blood sugar,insulin, HbA1c, glucokinase, glucose6-phosphatase, fructose-1,6-bis phosphatase, and lipid profile was estimated at the end of the study (45th day). The pancreas tissue was isolated and the changes was noticed. Statistical analysis: The data represent the mean ± standard deviation (S.D) of the indicated number of experiments. In the present investigation, since more than two treatment groups have been studied. Statistical analysis was performed using one way analysis of variance (ANOVA) by using statistical package of social science (SPSS) version 14.0 for windows. P values
