The Intervention Effect Of Jianpi Ruangan Decoction On NF-kB/TGF-?/Smads Signaling Pathway In Liver Fibrosis Rats

Objective Based on Yunnan Province Traditional Chinese medicine preferred treatment of liver fibrosis in the name of Jian Piruangan Decoction,this study will be carried out from the animal model to explore the mechanism of the party,intends to select the hepatic fibrosis indexes related to the occurrence and development of TGF-beta/Smad2/3 /NF-k B/Smad7/TNF-?,observeing these indicators to investigate the molecular mechanism of the the intervention of liver fibrosis TGF-beta signaling pathway.Methods1 Animal experiment: 90 male SPF rats?provided by the experimental animal center of Chengdu University of Traditional Chinese Medicine?were divided into 6 groups,normal group,CCl₄ model group,positive medicine Fuzheng Huayu capsule group,Jianpi Ruangan Decoction of low,medium and high dose groups,15 rats in each group.90 rats were fed after feeding for 1 weeks.2 Liver fibrosis rat model: the normal group was injected with normal saline,the other groups were injected intraperitoneally with 2ml/kg 70% olive oil and 30% CCl₄ mixture,about two times a week.3 Intragastric administration: drug prevention,from the model was established,the normal group and model group were given normal saline,positive drug group were treated with Fuzheng Huayu capsule orally?Clinical equivalent dose?,Jianpi Ruangan Decoction and experimental group according to the high?2.16g/kg/d?,medium?1.08g/kg/d?and low?0.54g/kg/d?dose administered 1times daily for 4 weeks.4 Index observation and detection:4.1Observation of Jianpi Ruangan decoction after treatment,the general condition of the rats during the experiment;4 weeks later rats were killed,serum and liver tissue,the amount of electronic weigh wet liver weight,liver index calculation.4.2HE staining and collagen fiber staining: the pathological changes of liver tissue in each group were observed by pathological section.4.3Serum biochemical parameters: the detection of ALT and AST in serum of rats in each group by the method of ELISA kit.4.4Detection of NF-kB/TGF-beta /Smads signaling pathway: used Western Blot to detect Jianpi Ruangan Decoction intervention after treatment in liver tissue NF-k B,/Smad2/3 phosphorylation,Smad7 protein expression,ELSIA detection of inflammatory factor TGF-beta,TNF-alpha.Results1 The animal model of hepatic fibrosis: compared with normal group model group rats of general morphology : hair darkening,loss of appetite,weight growth is slow,the liver wet weight increased significantly?P<0.05?,liver index was significantly increased?P<0.05?.The pathological model group of rat liver tissue sections of liver cell swelling significantly compared with the normal group,the proliferation of the connective tissue around the hepatic sinus,portal vein was part of the pathological appearance of connective tissue lobule,and even the formation of false lobules.The results showed that CCl₄ liver fibrosis model was successful.2 Serum biochemical detection :with ALT and AST in serum in the model group compared with normal group significantly increased?P<0.05?.3 The regulating effect of Jian Pi Ruangan Decoction on liver fibrosis rat model is as follows:3.1 Different measurement of Jianpi Ruangan Decoction and rats general morphological and pathological changes compared with model groups of rats: hair color,daily food intake,body weight growth,HE staining,and collagen fiber staining pathology showed reduced inflammatory lesions in liver tissue of rats,liver cell necrosis range decreases,pseudolobule reduced fiber less connective tissue hyperplasia,partial reversal.The results indicated that the prescription of invigorating spleen and softening liver has certain anti fibrosis effect.The experimental group increased with Jianpi Ruangan Decoction dose,rat hair color gradually bright,consumption has gradually increased,alleviate the damage to liver cells,high doses of Jianpi Ruangan Decoction group and cell injury in a reversal of the situation.3.2 The content of ALT and AST in the serum of rats in the treatment group was significantly lower than that in the model group?P<0.05?.Liver tissue ELSIA to detect jianpiruangan Decoction and group TGF-beta,TNF-alpha protein expression was significantly lower than the model group?P <0.05?.Proinflammatory cytokine of TGF-in liver tissue of Jianpi Ruangan Decoction group,alpha TNF-expression compared with Fuzhenghuayu capsule group decreased?P < 0.05?.3.3 Expression of NF-k B,Smad2/3,Smad7 protein of NF-k B/ TGF-beta 1/Smad2/3 signaling pathway: experiment of NF-k B,Smad2/3 phosphorylation of protein,Jianpi Ruangan Decoction group and Fuzheng Huayu capsule group,was significantly lower than those in model group?P <0.05?,Jianpi Ruangan Decoction is better than Fuzheng Huayu capsulegroup?P < 0.05?.The expression of Smad7,Jianpi Ruangan Decoction group and Fuzheng Huayu capsule group than in model group was significantly increased?P < 0.05?.The expression of Smad7 protein in Jianpi Ruangan Decoction group higher than the amount of Fuzheng Huayu capsule group?P <0.05?.Showed that the expression of Jianpi Ruangan decoction can inhibit NF-k B/TGF-beta/Smad2/3 signaling pathway.Jianpi Ruangan Decoction low,medium and high dose group increased with the dosage,liver tissue expression of TNF-alpha,TGF-beta,NF-k B and Smad2/3were decreased,Smad7 protein expression increased gradually;TGF-?,TNF-alpha,NF-k B and Smad2/3 increased the most obviously,the content of Smad7 is the most lowest in the high dose group Jianpi Ruangan Decoction.Shows that the treatment effect of high doses of Jianpi Ruangan Decoction on hepatic fibrosis best.Conclusion1 The prescription of Jipi Ruangan Decoction has the function of resisting CCl₄ hepatic fibrosis,and The best effect of high dose of Jipi Ruangan Decoction.2 Jianpi Ruangan decoction can achieve anti hepatic fibrosis by expression of protein intervention in the NF-k B/TGF-beta/Smads signaling pathway and the role of the high dose group of Jian Piruangan Decoction inhibition of this pathway is more obvious.