Discussed From PI3K/Akt/P70^s6K Signaling Pathway In Ethyl Acetate Extract Of Alysicarpus Vaginalis（L.）DC To Study The Mechanism In Anti-hepatic Fibrosis

Objective:To study the Anti-hepatic Fibrosis mechanism of acetate extract of Alysicarpus Vaginalis（L.）DC and the relationship with the PI3K/Akt/P70^s6K signaling pathway.Methord: Prepareing the acetate extract of Alysicarpus Vaginalis（L.）DC and lavage rats to get the medicated serum of acetate extract of Alysicarpus Vaginalis（L.）DC.Studying the influnence of medicated serum of acetate extract of Alysicarpus Vaginalis（L.）DC on the growth of HSC-T6 cell which is a vitro model of Hepatic fibrosis by the method of MTT.The mice were divided into blank control,model,Colchicine（0.2 mg·kg-1）,high-,mid-,and low-dose（the equivalent crude drug dosage were 40,20,and 10 g·kg-1）of the ethyl acetate extract of Alysicarpus Vaginalis（L.）DC groups.Each groups were induced by subcutaneous injection of CCL₄ in mice once every 3d and for 10 weeks,apart from the blank control mice.The mice were treated by intragastric administration the corresponding drugs or distilled water once a day and last for 10 weeks.Respectively test the content of ALT,AST in blood serum and the Col Ⅰ,Col Ⅲ in mice liver.Making pathological section and use HE,Masson staining to see the stage of mice liver fibrosis.For explain the relationship from the the Anti-hepatic Fibrosis mechanism of acetate extract of Alysicarpus Vaginalis（L.）DC and the PI3K/Akt/P70^s6K signaling pathway,we use the method of RT-PCR to detect the genetic transcription,and the Immunohistochemistry to detect the protein expression,which is about PDGF-BB,PTEN,PI3K,Akt,P70^s6K and α-SMA in the PI3K/Akt/P70^s6K signaling pathway.Resμlts: Compared with blank control group,The 10%,5%,2.5% of medicated serum of ethyl acetate extract of Alysicarpus Vaginalis（L.）DC significantly inhibit the growth of HSC-T6 cell（P<0.05）.Compared with model group,the high-,mid-,and low-dose of the ethyl acetate extract of Alysicarpus Vaginalis（L.）DC significantly decreased the activity of ALT and AST in serum of CCL₄-induced acute hepatic injury mice（P<0.05）,reduced the mortality of mice which was induced by subcutaneous injection of CCL₄.The high-,mid-dose of the ethyl acetate extract of Alysicarpus Vaginalis（L.）DC significantly decreased content of Col Ⅰ,Col Ⅲ in mice liver,decreased the the stage of mice liver fibrosis and have low inflammatory cell infiltrate.It still can decreased the hyperplasia of the liver portal area fiber connective tissue and grade of Ishak.The low-dose of the ethyl acetate extract of Alysicarpus Vaginalis（L.）DC is close to the model group,but it can decreased content of Col I in mice liver.The high-,mid-dose of the ethyl acetate extract of Alysicarpus Vaginalis（L.）DC significantly reduce the genetic transcription of PDGF-B,PI3K,Akt,P70^s6K,α-SMA,increase the genetic transcription of PTEN,reduce the protein expression of PDGF-BB,PI3K,Akt,P70^s6K,α-SMA,increase the protein expression of PTEN.The low-dose of the ethyl acetate extract of Alysicarpus Vaginalis（L.）DC can reduce the genetic transcription of PI3K,reduce the protein expression of PDGF-BB,increase the protein expression of PTEN.Conclusion: The ethyl acetate extract of Alysicarpus Vaginalis（L.）DC can significantly inhibit the growth of HSC-T6 cell,decreased the content of Col Ⅰ,Col Ⅲ in mice liver which was induced by subcutaneous injection of CCL₄.The Anti-hepatic Fibrosis mechanism of acetate extract of Alysicarpus Vaginalis（L.）DC probably through reduce the genetic transcription of PDGF-B,PI3K,Akt,P70^s6K,α-SMA,increase the genetic transcription of PTEN,reduce the protein expression of PDGF-BB,PI3K,Akt,P70^s6K,α-SMA,increase the protein expression of PTEN,which is in the PI3K/Akt/P70^s6K signaling pathway to reduce the stage of mice liver fibrosis.