Action of Corilagin on Hyperglycemia, Hyperlipidemia and Oxidative Damage in Streptozotocin-induced Diabetic Rats

ABSTRACT:

Diabetes mellitus is a world’s largest endocrine disease involving metabolic disorders of carbohydrate, protein and fat. This study was undertaken to investigate the anti-diabetic activity of corilagin, a member of polyphenolic tannins used against hyperglycemia and many other diseases in well-known animal models. Diabetes was induced chemically by intraperitoneal administration of Streptozotocin (40 mg/kg bw) to albino Wistar rats, which showed significant increase in the levels of fasting blood glucose, glycated haemoglobin, total cholesterol, triglyceride, low density lipoprotein cholesterol, very low density lipoprotein cholesterol, and a significant decrease in the level of body weight, plasma insulin, high density lipoprotein cholesterol, antioxidant activities, viz. superoxide dismutase, catalase and reduced glutathione. However, after 30 days of oral administration of corilagin (10and 20 mg/kg bw/day) to these diabetic rats evoked significant alterations in the above mentioned parameters. The efficacy of corilagin was compared with the standard glibenclamide (0.1 mg/kg body weight/day). Thus, our study demonstrates that corilagin has a potential to regulate diabetes, by exhibiting antidiabetic, antihyperlipidemic and antioxidant properties in STZ induced diabetic rats.

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Keywords

Corilagin; streptozotocin; glibenclamide; antidiabetic; antihyperlipidemic; antioxidant

1. Introduction

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The world’s largest endocrine disease involving metabolic disorders is diabetes mellitus (DM) (Chaiyasut et al. 2011). DM is characterized by hyperglycemia due to relative or absolute deficiencies in insulin action or insulin secretion. DM is considered a “modern day epidemic” and is recognized as a global public health issue due to its substantial financial burden worldwide on the healthcare system and its negative impact on the quality of life. Over the last century, there is a dramatic increase in the incidence of diabetes resulted from obesity, sedentary lifestyle and consumption of calorie rich diets (Souto et al. 2011). According to International Diabetes Federation (IDF), 382 million people (8.3%) suffer from DM and the estimation of this illness is projected to increase more than 592 million by 2035 worldwide (IDF, 2014).

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A large body of evidence suggests that inability of pancreatic β-cells to produce a physiologically appropriate amount of insulin leads to a chronic hyperglycemic condition in DM. The chronic hyperglycemia of diabetes generates auto oxidation of glucose and auto oxidative glycosylation of proteins which leads to oxidative stress by increasing the reactive oxygen species (ROS) (King and Loken, 2004). ROS contribute to the development of diabetic complications including tissue damage, heart and peripheral vascular complaints, retinopathy, nephropathy and neuropathy (Wiernsperger, 2003).

Drug prescription remains to be the major successful approach to improve diabetic condition, despite lifestyle modification as the first-line approach for an early stage of diabetes. Currently, different types of oral hypoglycemic agents are available along with insulin for the

treatment of DM. But these products possess adverse effects which include hypoglycemia, hepatotoxicity, dyslipidemia and attenuation of response after protracted use (Tahrani et al. 2011). In consequence, there is a need for the search of new therapeutic agents. Medicinal plants have always been rich sources of biologically active compounds vital to human health. Some plant-derived bioactive compounds, including epicatechin (Chakravarthy et al. 1982), quercetin (Shisheva and Shechter, 1992), pueranin (Hsu et al. 2003), kaempferol (De-Sousa et al. 2004), hesperidin (El-Alfy et al. 2005), proanthocyanidins and genistinin (Lee, 2006), are also known to regulate hyperglycaemia. Corilagin (Figure 1) (Schmidt and Lademann, 1951) is a member of polyphenolic tannins, has been discovered in a number of medicinal plants such as Punicagranatum(Guo et al. 2017),Terminaliachebula(Grover and Bala, 1992),Rosarugosa,Eugeniacaryophyllata,Punicagranatum(Li et al. 2014),Phyllanthusniruri(Jia et al. 2013),Emblicaofficinalis(Khan, 2009),DimocarpusLongan,Phyllanthusurinaria(Zheng et al. 2016), Syzygiumcumini(Chauhan and Intelli, 2015).Recent studies have shown that corilagin exhibit health benefits including antioxidant (Kinoshita et al. 2007), antihypertensive (Cheng, 1995), antiapoptotic and hepatoprotective effects (Hau, 2009). Corilagin also exhibit protective action against herpes simplex virus (HSV) 1 encephalitis (Guo et al. 2010). A previous study has shown that corilagin is protective against GalN/LPS-induced liver injury through suppression of oxidative stress and apoptosis (Kinoshita et al. 2007), and act as an inhibitor of TNF-α (Okabe et al. 2001). Furthermore, NF-κB activation also inhibited by corilagin (Gambari et al. 2012). The latest research reported that damage caused by cigarette smoke on lung epithelial cells could be attenuated by corilagin (Muresan et al. 2015). Although, there is an association exists between hyperglycemia  and  corilagin  (Honma  et  al.  2010),  there  is  no  systematic  investigation  is

available on anti-diabetic activity of corilagin. Therefore, the present work was carried out to evaluate the efficacy of corilagin in regulating DM.

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Figure1:Strcture of corilagin.

2. Materials and methods

1. Materials

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Corilagin (≥95%) and STZ were purchased from Sigma- Aldrich (St. Louis, MO). For the biochemical assessments, the diagnostic kits were supplied from Swemed Biomedicals Pvt. Ltd.

(Bengaluru, India). All the other chemicals used in the present study are of analytical grade obtained from standard commercial suppliers.

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2. Maintenance of animals

Healthy adult albino Wistar rats were used in the current study, weighing 180–210 g was acquired from Central Animal House, Department of studies in Zoology, University of Mysore. The animals were housed in standard polypropylene cages and maintained in an air-conditioned room (25 ± 10C) with a 12-h light/12-h dark cycle and were fed a standard diet of known composition, and water adlibitum.Animal experiments were performed in accordance with regulations specified and monitored by the Institutional Ethics Committee of University of Mysore (Approval number – UOM/IAEC/18/2012) and followed the guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals.

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3. Induction of experimental diabetes

Streptozotocin (STZ) was freshly dissolved in (0.01 M, pH 4.5) citrate buffer. DM was induced in overnight fasted rats by a single intraperitoneal injection of STZ (40 mg/kg b.w.), and the injection volume was 1 ml/rat. Control animals were injected with citrate buffer alone. After 72h of STZ injection, diabetes was confirmed by measuring the fasting blood glucose (FBG) concentration. The animals with FBG values above 235 mg/dL were considered to be diabetic and included in the present study (Prisilla et al. 2012).

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4. Experimental design

Rats (n=25) were randomized into the following groups after the induction of STZ diabetes:

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Group I: Control

Group II: Diabetic Control

Group III: Diabetic + Glibenclamide (0.1 mg/kg body weight/day) Group IV: Diabetic + Corilagin (10 mg/kg body weight/day) Group V: Diabetic + Corilagin (20 mg/kg body weight/day)

Corilagin and glibenclamide were diluted in water and administered via oral intubation daily for a period of one month. The initial and final body weights of each rat in various groups were recorded. At the end of the experimental period, all the animals were fasted overnight, and sacrificed under mild ether anesthesia.

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Blood samples were collected from the carotid artery and centrifuged at 1000 rpm for 10 min, and the serum was obtained and used for various biochemical estimations.

5. Analytical procedure

1. Fasting blood glucose (FBG)

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At weekly intervals, blood samples were withdrawn by nicking the tip of the rat tail and the FBG level was estimated using glucometer (Accu-Chek, Mannheim, Germany) in which its principle is based on the glucose oxidase method. Results were expressed as mg/dL.

2. Plasma insulin and glycated hemoglobin (HbA1c)

Plasma insulin was estimated using a commercial ELISA kit purchased from DRG diagnostics (GmbH, Germany). HbA1c in whole blood was measured using commercial assay kit following the protocol provided by manufacturer (Swemed Biomedicals, Pvt. Ltd., Bengaluru, India).

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3. Measurement of Serum Lipid Profile

Serum concentrations of triglyceride (TG), total cholesterol (TC), and high-density lipoprotein cholesterol (HDL-C) were determined through Artos –Versatile Clinical Chemistry Analyzer using commercially available kits (supplied by Swemed Biomedicals, Pvt. Ltd., Bengaluru, India). Low-density lipoprotein cholesterol (LDL-C) and very  low-density lipoprotein cholesterol (VLDL-C) were calculated according to Friedewald’s formula LDL = [(TC − HDL) − TG/5] and VLDL cholesterol =TG/5 (Friedewald et al. 1972).

6.                        Measurements of antioxidant activity

1.    Assay of Superoxide dismutase (SOD)

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Inhibition of superoxide driven oxidation of quercetin by SOD at 406 nm was measured in this assay. The complete reaction mixture consisted of 25 mM phosphate buffer (pH 7.8), 0.25 mM EDTA, 0.8 mM TEMED and 0.05 μM quercetin. The amount of enzyme that inhibits the auto-oxidation of quercetin by 50% was defined as one unit (Kostyuk and Potapovich, 1989).

2.    Assay of Catalase (CAT)

CAT activity was estimated by Aebi method (1984), following the clearance of H2O2 at 240 nm in a reaction media containing 50 mM phosphate buffer (pH 7.0), 0.5 mM EDTA, 10 mM H2O2, 0.012 %TRITONX100. Activity was expressed as µmol H2O2 (HP) decomposed/min/mg protein.

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3.    Assay of reduced glutathione (GSH)

Samples were taken in 96-well microtiter plates. The final volume was made up to 100

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µL with 100 mM phosphate buffer (pH 8.0, containing 5 mM EDTA), and 25 µL O- phthalaldehyde was added and incubated for 45 min at 37° C. After exciting the samples at 340 nm, fluorescence emission was recorded at 425 nm and the GSH was calculated (Hissin and Hilf, 1976).

2.7. Statistical analysis

Data were analyzed by one way analysis of variance followed by Duncan’s multiple range test using SPSS software package, version 14 (Chicago, IL). Results were expressed as the mean ± SE. The P-value <0.05 was taken as indicating statistical significance.

3. Results

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Table1.Effectofcorilaginonbodyweightinstreptozotocin-induceddiabeticWistarrats.