Effect And Mechanism Of Schisandrol B Combined With Wedelolactone On Liver Fibrosis

Objective: To investigate the protective effects of Schisandrin(Schisandrol B,Sch-B)combined with pyrethrin(Wedelolactone,Wed)on hepatic fibrosis induced by bile duct ligation and carbon tetrachloride in mice.Method:1.use bile duct ligation(bile duct ligation,BDL)surgery to induce the establishment of a mouse model of cholestatic liver fibrosis.C57BL/6 mice were randomly divided into sham operation group(control),bile duct ligation model group(BDL-Model),colchicine positive drug group(colchicine 0.2mg/kg),Wed20mg/kg group,Sch-B40mg/kg group,and Wed20mg/kg Sch-B40mg/kg group.The sham operation group opened the abdominal cavity and stripped the total bile duct,but did not perform bile duct ligation,and the other four groups performed ligation after stripping the total bile duct.Postoperative d 7 began administration,continuous administration of 14 d..The contents of hydroxyproline(hydroxyproline,Hyp)in serum were detected by colorimetry;histopathological changes of liver were observed by HE staining,Masson staining and Sirius red staining(sirius red staining);expression of actin in liver tissue α-smooth muscle was detected by immunohistochemical staining;expression of fibroblast-promoting growth factor and proliferation chemokine m RNA in liver tissue was detected by real-time quantitative PCR method;and expression of TGF-β /Smad signaling pathway and NF-κB signaling pathway-related protein in liver tissue was detected by immunoblotting.2.use of CCL4 to induce liver fibrosis in mice with chemical liver injury.C57BL/6 mice were randomly divided into olive oil solvent control group(control),ccl4 model group,colchicine positive drug group(colchicine0.2mg/kg),Wed20mg/kg group,Sch-B40mg/kg group,and Wed20mg/kg Sch-B40mg/kg group.The CCL4 was dissolved in olive oil and prepared into5% CCL4 oil solution with 2 m L/kg.intraperitoneal injection 1 time a day,14 d after the start of treatment medication.After 28 d of treatment,blood and tissue samples were taken.the detection index is same as above.3.Inflammatory stimulation model and fibrosis model were established in vitro to investigate the anti-fibrosis mechanism of Wed and sch-b.Results:1.compared with the model group,both Wed and Sch-B groups had obvious liver protection effect,and could also reduce the serum Hyp content.2.HE staining,Masson staining,sirius red staining sho Wed that compared with the model group,WED and Sch-B administration alone could reduce the proliferation of small bile ducts in the liver tissue of mice and reduce the formation of inflammatory cell infiltration and fibrous septum.the combined administration group had the best effect and had significant difference.The proliferation of hepatic stellate cells was inhibited by3.WED and Sch-B administration alone and in combination.When compared with the model group,the expression of Collagen 1 protein in liver tissue was reduced by WED and Sch-B administration alone and combined administration group.4.WED and Sch-B administration alone inhibited transcription of TGF-β /Smad signaling pathways.The expression of Smad2/3 protein was down-regulated in WED and Sch-B groups.5.WED and Sch-B alone,combined administration and combined administration can inhibit the transcription of NF-κB signaling pathway.When compared with the model group,WED and Sch-B administration group alone and combined administration group could reduce the expression level of P-I κ B α protein and the combined administration group had the strongest effect,and there was significant difference.6.Wed mainly inhibited activation of NF-B signaling pathway and inflammation in macrophages,and reversed liver fibrosis in hepatic astrocytes by inhibiting TGF-TGF /smads signaling pathway.Sch-b has synergistic effect with Wed.Conclusion:Wed and Sch-B can effectively block and inhibit the formation of BDL and CCL4 induced liver fibrosis in mice.the mechanism may be related to its inhibition TGF-β /Smad and transcription of the NF-κ B signal transduction pathway.