| ObjectiveThis experiment adopts the high fat forage feeding rats,to establish nonalcoholic fatty hepatitis model,to observe the effects of Jianpishugan Decoction on liver pathology,liver function,serum lipid,insulin resistance and oxidative stress in rats with nonalcoholic steatohepatitis,to investigate the mechanism of jianpi shugan Decoction in the treatment of nonalcoholic steatohepatitis,for the clinical treatment of traditional Chinese medicine to provide laboratory basis nonalcoholic fatty hepatitis.MethodsAccording to the random number table method,40 SPF grade rats were divided into 6 groups:blank group,model group,low dose group,medium dose group,high dose group and polyene phosphatidylcholine group.Except the model group,there were 6 in each group.The rats were modeled after 1 week of adaptive feeding.The blank group was fed normal diet,and the other groups were fed high-fat diet for 8 consecutive weeks.After successful modeling,rats fed high-fat diet were randomly regrouped and each group was fed normal diet,the model group and the blank group were reperfusion treated with deionized water at 1ml/100g,the dosage groups of Jianpishugan Decoction and yishanfu group were administered with Jianpishugan Decoction and yishanfu suspension by gavage respectively.The equivalent dose of this experiment was set as the medium dose group of Jianpishugan Decoction(dosage:15.05g/kg),the high dose group was given at 3 times the concentration of the medium dose group,the low dose group was administered at 1/3 the concentration of the medium dose group,dosage of yishanfu group:137.3mg/kg,(the experimental dose is calculated according to the equivalent dose ratio table of the conversion of human and animal body surface areas),the intervention lasted for 4 weeks,once a day,during which free drinking water and diet were observed,the animal’s mental state,hair,stool and drinking water were observed,and the body mass was weighed once a week.At the end of the 12th week of the experiment,rats in each group were given a fast of 24 hours after gavage.After weighing,10%chloral hydrate was used as an intraperitoneal injection for anesthesia(0.3ml chloral hydrate was used for every 100g of the weight of the rats).After the effect of anesthesia,blood from the abdominal aorta of rats was rapidly taken,and the blood samples were placed at room temperature for 2 hours before centrifugation at a speed of 2000 rotations per minute.Then take the supernatant-80℃ refrigerator,completes the date marking to be detected,detection of serum alanine aminotransferase(ALT),serum aspertate aminotransferase(AST),serum triglycerides(TG),serum total cholesterol(TC),serum high-density lipoprotein(HDL),serum low density lipoprotein(LDL),insulin(FINS),glucose(FPG),the levels of serum adiponectin(ADPN)and according to the FINS and FPG numerical calculation of insulin resistance(HOMA IR)level,clear between groups on the liver enzyme,blood lipids and other indicators.The liver was quickly removed after blood collection,and the wet weight of the liver was observed,and the liver index(wet weight of the liver/total weight X 100%)was calculated.Then,from the right lobe of the liver,the size was about 0.5cm X 0.5cm X 0.5cm,4%paraformaldehyde solution was put into the fixed sample,and the date was marked,and the sample was put into the refrigerator for detection.After the supernatant was taken from the liver tissue homogenate,the levels of superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA)were detected.Results①General conditions and pathological morphology of the liver:compared with the normal group,the rats in the model group had brown and dull hair color,happy to reunite and curl up,watery and odorous stools,and significantly increased NAS pathological score and liver index.Compared with the blank group,there were no obvious abnormalities in the hair color,behavior,diet and excreta of rats in each group of Jianpishugan Decoction and yishanfu group.Compared with the model group,the liver index of the middle and high dose groups of Jianpishugan Decoction and the yishanfu group were significantly lower.Compared with the model group,the medium and high dose groups of of Jianpishugan Decoction and the yishanfu group could significantly reduce the steatosis and balloon-like variation scores of the model group,and the Jianpi shugan prescription group and yishanfu group could significantly reduce the inflammation in the lobules.②Liver enzymology:serum ALT and AST in the model group were significantly higher than those in the normal group.In the middle and high dose groups of Jianpishugan Decoction and the yishanfu group,the ALT and AST levels were significantly reduced,while the low dose had no significant effect on the level of liver enzymes.③Blood lipid:serum TG,TC and LDL in the model group of Jianpishugan Decoction were significantly higher than those in the normal group,while HDL was not significantly lower.In the high-dose group of Jianpishugan Decoction and yishanfu group,the TG level was significantly reduced;in the low-dose and medium-dose groups of Jianpishugan Decoction,the TC level was significantly reduced;in the low-dose,high-dose and yishanfu groups,the HDL level was significantly increased.The medium dose group and the group of yi shanfu could significantly reduce the level of LDL.④oxidation and antioxidant:the levels of SOD and CAT in the liver tissue of the model group were significantly lower than those of the blank group,and the levels of MDA were significantly higher.SOD levels were significantly increased in the low-dose,high-dose and yishanfu groups.CAT levels were significantly increased in the middle and high dose groups and the yishanfu group.The levels of MDA were significantly reduced in each group of Jianpishugan Decoction and yishanfu group.⑤insulin resistance:the FINS,FPG and Homa-IR levels of the model group were significantly higher than those of the normal group,while the ADPN was significantly lower.The high dose group and yi shanfu group could significantly reduce the FINS level.FPG level was significantly reduced in the middle and high dose groups.Each intervention group can significantly improve the insulin resistance of rats.The ADPN level was significantly increased in the middle and high dose groups of Jianpishugan Decoction and the yi shanfu group.ConclusionJianpishugan Decoction can effectively improve the pathological state of the liver tissues of NASH rats,effectively improve the liver cell steatosis and inflammatory response,by regulating the lipid metabolism of rats,reducing insulin resistance,inhibiting lipid peroxidation and improving the antioxidant capacity of the rat liver,reduce the level of liver enzymes and thereby reduce the damage of rat liver cells. |
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