Effect Of Shugan Lipi Recipe On Intestinal Mucosal Permeability Of Rats With Non-alcoholic Steatohepatitis

Objective:Based on the "enter-liver axis",this study tested the permeability of intestinal mucosa to demonstrate the effective mechanism of shugan lipi recipe on NASH rats,providing experimental basis and theoretical support for the effective prevention and treatment of NASH with traditional Chinese medicine and providing certain clinical guidance.Methods:Sixty SD male rats were randomly divided into 8 for the normal control group(group A)and 52 for the model group.After 1 week of adaptive feeding,model group rats were fed with MCD feed to replicate NASH model.At 9 weeks,4 model group rats were randomly selected for liver samples for HE staining to evaluate the success of modeling.48 only then rest model group rats were randomly divided into model + normal diet,no Chinese medicine group(group B),the model + normal diet + low doses of traditional Chinese medicine group(group C),the model of normal diet + + high dose of Chinese traditional medicine group(group D)+ + MCD diet without Chinese traditional medicine group and model group(E),model + MCD diet plus low dose of Chinese traditional medicine group(group F)+ MCD diet + high dose of Chinese traditional medicine group and model group(G).The drug was administered by gavage at week 9 for a total of 4 weeks.All rats were sacrificed after 12 hours of fasting after the last administration,and blood,liver and small intestine tissues were collected.Serum ALT,AST,TC and TG were determined by biochemical analyzer.Serum TNF-,il-6 and il-1 were determined by ELISA.Serum endotoxin was detected by limulus limulus reagent.HE staining was used to determine the liver pathology;transmission electron microscopy was used to observe the ultrapathology of the small intestine mucosa;fluorescence quantitative PCR and western-blot were used to observe the expression of TLR4 in the liver tissue and the expression of occludin and zo-1 in the small intestine mucosa tissue.Results: 1.All model group compared with normal control group,serum ALT,AST,TC,TG levels are rising,after diet or high dose of TCM intervention the ALT,AST,TC,TG levels decreased(P < 0.05),and diet in combination with high dose of Chinese medicine intervention had the greatest reduction(P < 0.01),low doses of intervention,though there is a downturn in the Chinese traditional medicine but without statistical significance;2.Compared with the normal control group,il-1,il-6,TNF-,ET were significantly increased in all model groups,and il-1,il-6,TNF-,and endotoxin were decreased after diet or high-dose Chinese medicine intervention(P<0.05).The decrease was most obvious when diet combined with high-dose Chinese medicine intervention(P<0.01).Low-dose Chinese medicine intervention showed a downward trend,but no statistical significance.3.Compared with the normal control group,hepatocyte steatosis and inflammatory cell infiltration were observed in the liver of all model groups by HE staining,which were alleviated by diet and high-dose drug intervention,among which the liver pathology was improved more significantly by diet combined with high-dose Chinese medicine intervention.Tem observation of intestinal mucosa showed that the intestinal mucosa of all rats in the model group became shorter and the desmosome structure relaxed,etc.The morphology and integrity of intestinal mucosa were improved after diet and high-dose drug intervention,among which the morphology and integrity of intestinal mucosa were better after diet combined with high-dose Chinese medicine intervention.4.Compared with the normal control group,the expression of TLR4 in the liver tissue of the model group was up-regulated,and the expression was down-regulated after the intervention of diet or high-dose Chinese medicine(P<0.05).The down-regulated expression was most obvious when diet combined with high-dose Chinese medicine intervention(P<0.01).Occludin and zo-1 were down-regulated in the model group.The expression was upregulated after the intervention of diet or high-dose Chinese medicine(P<0.05),and the upregulated expression was the most obvious when diet combined with high-dose Chinese medicine(P<0.01),while the upregulated expression was not statistically significant after the intervention of low-dose Chinese medicine.Conclusion:1.diet or high dose liver spleen alone the intervention in reducing the NASH rats liver inflammation and improve liver steatosis,intestinal mucous membrane permeability have curative effect,its mechanism may be through the intestinal mucosa is closely connected to the expression of the protein,which regulates intestinal mucous membrane permeability,reduce the serum IL-1,IL-6,TNF alpha,the level of endotoxin,inhibit the expression of TLR4 in order to reduce secondary liver injury;2.after the intervention of low-dose traditional Chinese medicine alone,the inflammatory indicators showed a decreasing trend,but there was no statistical significance,and the liver pathology and intestinal mucosal permeability were not significantly changed.