Danshensu Pi3-k/akt Signaling Pathway In Hepatic Stellate Cells

Aim: To isolate and get high output and purity hepatic stellate cells of rat.Method: The rat hepatic stellate cells were isolated with collagenaserecirculating perfusion and purified with density gradient centrifugation.Results:In this way,the output of the hepatic stellate cells amounted to 5-6×107 cells per rat. The purity of the cells was above 97%. The viability of the cells was more than 92%.Conclusion: Getting high output and purity hepatic stellate cells. Aim: To investigate the effect of Danshensu on the expression of Akt and its phosphorylated proteins (p-Akt) in Hepatic satellite cells (HSCs).Methods: HSCs were isolated and cultured from the liver of Wistar rat by in–situ perfusion with collagenase and density gradient centrifugation with Nycodenz. Then the HSCs were exposed to 0.0625mmol/L,0.125mmol/L,0.25mmol/L Danshensu and 20nmol/L,40 nmol/L,60 nmol/L Wortmannin for 24 hours. The effect of Danshensu and Wortmannin on cell proliferation was measured by MTT assay. The cell cycle of HSCs was analysed by flow cytometry. The expression of Akt and p-Akt were measured by Western blot.Results: At the concentration of 0.0625mmol/L, 0.125mmol/L, 0.25mmol/L, Danshensu could significantly inhibit HSC prolifer action in close-dependent manner. At the concentration of 20nmol/L, 40 nmol/L,60 nmol/L Wortmannin could significantly inhibit HSC prolifer action in close-dependent manner. Both Danshensu and Wortmannin could increase the numbers of HSCs in G1 phrase and decrease the numbers of HSCs in S phrase. Danshensu and Wortmannin had no marked influences on Akt expression. But p-Akt was expressed in HSCs, which was decreased with the increasing of the concentrations of Danshensu and Wortmannin.Conclusion: The anti-fibrotic mechanism of Danshensu might be associated with the decreased expression of p-Akt in HSCs.