Protective effect of Piper betle leaf extract against cadmium-induced oxidative stress and hepatic dysfunction in rats King Saud University Saudi Journal of Biological Sciences

Introduction Exposure Agricultural soils Mining activities of mines Contaminated plant sources Human activities Global cycle of heavy metals & metalloids Toxic non-essensial element Mining activities of mines Exposure Accumulates in biological system Injury to organs Disfuction organs

Ethnomedicines in India Bioactive molecules polyphenol Antioxidative property alkaloids steroids saponin tannins Piper betle Piper betle Ethnomedicines in India Low cost Easy accesibility to everyone Perceived fewer side effects The possible hepatoprotective activity of Piper betle leaves against Cd induced hepatotoxicity has not been reported so far

Objective To evaluate the hepatoprotective effect of the P.betle leaves by using the Cd-induce sub-chronic oxidative liver injury model in rats

Materials and methods

2-thiobarbituric acid (TBA) Butylated hydroxytoluene (BHT) Chemicals Cadmium chloride 2-thiobarbituric acid (TBA) Butylated hydroxytoluene (BHT) Reduced glutathione (GSH) 2,2-dipyridyl Xylenol orange 2,4-dinitrophenylhydrazine (DNPH) γ-glutamyl-p-nitroanilide 5,5-dithiobis-2-nitrobenzoic acid

Plant material & ekstrak preparation Piper betle Plant material & ekstrak preparation Preparation of ethanolic extract of Piper betle Dosage fixation Determination of total phenolic & flavonoid contents

Animals & experimental design Male albino Wistar rats Body weight 180-200 g The rats were allowed standar pellet diet and water adlibitum for the duration of the experiment Animals & experimental design 24 ekor 6 ekor G1 G2 G3 PBE (200 mg/kg/day) Normal saline G4 Cd (5 mg/kg/day) Cd (5 mg/kg/day) + PBE (200 mg/kg/day)

Activities of serum marker enzymes Biochemical assays Activities of serum marker enzymes Determination of lipid peroxidation & oxidative stress markers Determination of non-enzymatic & enzymatic antioxidants Estimation of membrane-bound ATPase

Histopathological studies Tissue samples Sections of tissues (5-6 µm thick) Fixed in 10 % formalin-saline Prepared by using a rotary microtone Dehydrated by passing (mixture of ethyl alcohol, water) Stained with hematoxylin & eosin dye Cleaned in xylene Which was mounted in a neutral deparaffine xylene medium for microscopical observations Embedded in paraffin

Statistical analyses Data were analyzed by one way analysis of variance (ANOVA) followed by Duncan’s mutiple range test (DMRT) using commercially available statistics software package (SPSS for Windows, V.13,0 Chicago, USA). Result were presented as mean ± SD. p value < 0,05 were regarded as statistically significant.

Result and discussion

Effects on body weight gain, relative liver weight, feed and water intake

Effects on serum hepatic marker enzymes and bilirubin Lanjutan

Effects on hepatic oxidative stress markers Ns

Effects on hepatic non-enzymatic antioxidants Ns

Effects on hepatic enzymatic antioxidants Ns

Effects on hepatic membrane bound ATPases Ns

Histological examination of liver tissue A : Control B : Cd treated rats C : Cd treated rats D : Cd+PBE treated rats E : PBE treated rats

Conclusion Cd is capable of causing marked oxidative stress in addition to deplete the antioxidants and inhibiting the activities of antioxidants enzymes. The treatment with PBE could significantly attenuate the Cd induced oxidative hepatotoxicity and shows its therapeutic potential to be used as a cost effective safe herbal oxidative agent in the treatment of Cd toxicity.