Protective Effects Of Salvia Miltiorrhiza Extract On Beta-Cypermethrin-Induced Liver Injury In Jian Carp

The liver is the main metabolism and detoxification organ of fish,its physiological function in the body is very important.Many compounds,including chlorinated solvents and drugs,can cause liver damage through their metabolic conversion to highly reactive substances and the generation of free radicals,therefore it is most vulnerable to a variety of poisons,metabolic toxin and pathogens.Presently in China,a fish liver disease called "fish hepatobiliary syndrome",with the symptoms of color change and enlargements of liver and gall bladder,has been frequently reported and caused dramatic losses in many cultured species.There have been no effective treatments for the fish hepatobiliary syndrome,hence the search for plant-based remedies is of considerable interest to researchers and aquatic medicine producers.The present study aimed to establish an in vitro beta cypermethrin(β-CYP)-induced hepatic injury model using precision-cut liver slices(PCLS)and study the hepatoprotective effect of Salvia miltiorrhiza extract in Carp.The main research contents and results are as follows:1)Acute toxicity of P-CYP:fish were exposed to β-CYP at 0,4.5,9,18,36,72 μg/L.Mortality was recorded at 24 h,48 h and 96 h.The calculated 96 h LC50 value of β-CYP was 26.53 βg/L.2)In vitro(precision-cut liver slices)β-CYP-induced hepatic injury model establishment.The precision-cut liver slices were exposed to p-CYP at 0.15,0.3,0.6,1.2 and 2.4 mg/mL.After 4 fours,the slices and supernatant were collected for biochemical analyses.The results showed that β-CYP at 0.6 mg/ml resulted in significant differences in glutamatepyruvate transaminase(GPT),glutamate oxalate transaminase(GOT),alkline phosphatase(AKP),lactate dehydrogenase(LDH),superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and catalase(CAT),total antioxidant capacity(T-AOC),while slices treated with β-CYP at 0.6 mg/ml showed little difference with control group in adenosine triphosphate(ATP)content.Therefore,β-CYP at 0.6 mg/ml was selected as the inducer for the model establishment.3)Protective effects of Salvia miltiorrhiza extract(SME)against liver injury in vitro:Salvia miltiorrhiza extract(0.2,0.4 and 0.8 mg/ml)was added to PCLS culture system before(pre-treatment),after(post-treatment)and both before and after(pre-and post-treatment)the exposure of PCLS to 0.6 mg/ml β-CYP.The supernatants and PCLS were collected for biochemical analyses.Results showed that compared with the control,Salvia miltiorrhiza extract in post-treatment group and pre-and post-treatment group displayed the better effect on the protection of liver slices than the per-treatment group.Salvia miltiorrhiza extract at 0.4 and 0.8 mg/ml significantly reduced the levels of GOT,GPT,AKP,LDH and MDA,increased the activities of antioxidant enzymes such as SOD,T-AOC,CAT,GSH-Px,GST,and increased the contents of TP,Alb and ATP in liver slices.Conclusion:1)The 96 h LC50 value of β-CYP was 26.53 μg/L.2)The suitable concentration of P-CYP for hepatic injury model was 0.6 mg/ml.3)Salvia miltiorrhiza extract can effectively reduce the fish liver damage by β-CYP,it may potentially be used as a hepatoprotective agent for fish liver disease.