The Influence Of HIT For Liver Fibrosis In Rats Of The TLR4-NF-?B Signal Transduction Pathways

Objective: On the basis of the preliminary study further in CCl4 induced liver fibrosis in rats as the object of study,using ELISA and fluorescence quantitative PCR and IHC of method of trefoil HIT of TLR4-NF-?B signal transduction pathway to clear HIT whether through the regulation TLR4-NF-?B signal transduction pathway and achieve the role of anti liver fibrosis.This project through the study of HIT of TLR4-NF-?B signal transduction pathway of influence,is expected to in signal transduction molecular level clear anti hepatic fibrosis mechanism of HIT,HIT anti-hepatic fibrosis provides new theoretical basis.Methods:1.HIT extract extraction,enrichment,the suction filter,water extract-alcohol precipitation mainly adopts the method of extraction,the concentrate filtering,collect and filtrate concentrated into concrete;2.Experimental animal grouping and building: 106 healthy clean grade SD rats,male and female 50,140¹80 grams of weight.Randomly divided into 6 groups and blank control group,the positive group,HIT model group,HIT high dose group,HIT medium dose group,HIT low dose group.HIT high dose group?80g/kg,according to the content similarly hereinafter?,HIT medium dose group?40g/kg?,HIT low dose group?20g/kg?,12 weeks of treatment.Blank group is given ordinary feed,free drinking water.Model Groups,all give back subcutaneous injection of 40% CCL₄ peanut oil solution,according to the 0.3ml/100 g,injection once every 3 days,at the same time HIT dosage groups and positive group to are given stomach perfusion,and blank model group give water to fill the stomach every day,a total of 12 weeks,success after inspection,take blood profile control liver;3.Using the method of ELISA to detect the expression of AST,ALT,IL-6,IL-1 and TNF-?.4.Using the fluorescent quantitative PCR method to detect the expression of TLR4 and My D88 in the liver tissue;5.Using the IHC detection of liver tissue NF-? B protein expression;Results: 1.The model group rats 12 weekends body weight,viscera coefficient of liver were significantly higher than that of blank group?P<0.01?,there is a significant difference.Positive group body weight com-pared with model group had obvious difference?P<0.05?,the liver is obviously better than the coefficient of model group?P<0.01?,there are significant differences,HIT a high dose group,middle dose group liver coefficient is better than that of model group?P<0.05?,there are differences;HIT a low dose group compared with model group has no statistical significance.2.The model group rat serum ALT,AST were abnormally elevated,respectively,compared with the margin group,the results had signify-cant difference?P<0.01?.Compared with the model group,the positive group?P<0.01?,and HIT the high dose group?P<0.05?,dose?P<0.01?in the HIT and HIT a low dose group?P<0.05?can significantly reduce liver fibrosis in rats serum ALT level.Compared with the model group,the positive group?P<0.01?,and HIT the high dose and HIT medium dose group?P<0.05?can reduce hepatic fibrosis in rats serum AST expression level,HIT a low dose group and model group than there is no significant difference?P>0.05?,no statistical significance.3.The model group rats serum IL-1?,IL-6,TNF-? are unusually high,respectively,compared with the margin group,the result had significant difference?P<0.01?.Compared with the model group,the positive group?P<0.01?,and HIT the high dose group?P<0.05?,can significantly reduce liver fibrosis in rats serum IL-1? expression,dose in the HIT and HIT a low dose group of lower IL-1? ratio is larger than the model group,but there is no statistical significance.Compared with the model group,the positive group,HIT a high dose?P<0.05?can reduce hepatic fibrosis in rats serum IL-6expression level,HIT the dose group?P<0.01?can significantly decrease the liver fibrosis in rats serum IL-6expression level,HIT a low dose group of lower IL-6 ratio is larger than the model group,but there is no statistical significance.Compared with the model group,the positive group?P<0.01?can significantly reduce the expression level of liver fibrosis rats serum TNF-?,HIT the dose group?P<0.05?can reduce hepatic fibrosis in rats serum TNF-?expression level,HIT a high dose group,low dose group reduce TNF alpha ratio is larger than the model group,but there is no statistical significance.4.Model group TLR4 m RNA expression in rat liver tissue relative blank group increased significantly?P<0.01?;HIT high dose treatment group and positive group compared with model group TLR4 m RNA expression significantly reduced?P<0.01?;HIT therapy group,low dose group TLR4 m RNA expression under positive group,there were significant differences?P<0.05?.In the model group rat liver tissue My D88 m RNA expression of relative blank group increased significantly?P<0.01?.HIT low dose treatment group compared with model group My D88 m RNA expression was significantly reduced?P<0.01?,positive group compared with model group My D88 m RNA expression reduced?P<0.05?.Compared with positive group,HIT lower dose treatment groups My D88 m RNA expression of higher than that of positive group,compared with significant differences?P<0.01?;HIT high dose treatment group My D88 m RNA expression of higher than that of positive group,compared with the difference?P<0.05?.5.IHC results:Model group compared with the margin group,NF-? B significantly increased?P<0.01?,liver cells,hepatic sinus,portal area around blood vessels and bile duct wall around the deep cells visible positive staining area,accidentally visible flake fusion zone,coloring.Compared with model group,positive group of NF-?B protein expression level decreased significantly?P<0.05?;HIT high dose group,middle dose group and low dose group of NF-? B protein expression level decreased obviously?P<0.05?,shaded area part into small flake and a small amount of light yellow color cells.Conclusion: 1.HIT can reduce the liver fibrosis in rats serum ALT,AST,IL-1?,the expression of IL-6 and TNF-? level,clew HIT with lower disease factor expression of function,the effect of anti fibrosis may be related to regulating IL s and TNF-a,and then reduce the secretion of extracellular matrix,balance the secretion and degradation of collagen in the liver.2.HIT can significantly reduce hepatic fibrosis in rats of TLR4,My D88 expression,can reduce the expression of NF-?B protein,can regulate TLR4-NF-?B signal transduction pathway and protein expression was restricted,HSC activation is further inhibited,reduce the secretion of extracellular matrix,thus the degree of liver fibrosis reduce.