Experiment Study On Akt Inhibited The Hepatic Stellate Cell Activation

Part one:Study on method of inducing cirrhosis model in rats by carbontetrachlorideObjective: To establish a reliable rat model of liver cirrhosis for next study and to investigate the change of the expression ofα-SMA in liver of rats with liver cirrhosis. Methods: 90 male SD rats were randomly divided into two groups,The experiment group(80) were treated with 40% CCL₄ by subcutaneous injection two weeks,then the concerntration of CCL₄ was adjusted to 50%,the dose of CCL₄ was adjusted according to the weight changes,twice a week,keep on 8 weeks total.The control group(10) were injected with normal saline,the frequency dose and period as same as the experiment group,liver biopsy after 8 weeks,HE Massion trichrome staining were played with liver tissue,the expression ofα-SMA were measured with immunohistochemistry.Results: After being induced with above means,the rat mortality of the experiment group was 11.25%,and the formation rate of liver cirrhosis false lobule was 88.75%. Compared with the control group,the expression ofα-SMA in livers was significantly stronger in the experiment group.the control group(10) were survival.Conclusion: The methods that subcutaneous injected with CCL₄ can established reliable rat model of liver cirrhosis,α-SMA play a role in the development of liver cirrhosis.PART TWOExperiment study on of Akt inhibited the hepatic stellate cell activation in rats. Objective: To investigate whether ntra-janitrix administration of reorganization plasmid vecter for pcDNA3.1/Akt could inhibit the HSCs activation to a rat model of liver cirrhosis by observe the diversity of the collagen deposition and the expression ofα-SMA after intra-janitrix administration of reorganization plasmid vecter for pcDNA3.1/Akt.Methods: Sixty Sprague-Dawley rats of liver cirrhosis were divided into three groups: Akt-treated group(n=20),control plasmid-treated group(n=20) and Tris buffer-treated group(n=20).Liposome-Akt was transduced into rat of Akt-treated group through the injection to portal vein;group; control plasmid-treated group and Tris buffer-treated group were received control plasmid or Tris buffer respectively. the diversity of collagen depositon was observed by Massion trichrome staining and the expression ofα-SMA in liver was measured by immunohistochemistry (SABC) after executed the rats.Result: Compared with the control plasmid-treated group or Tris buffer-treated group,liver histology showed that architecture of liver got recovery false lobules were resolution the collagen deposition decreased and the expression ofα-SMA in liver was significantly decreased in the Akt-treated group.Conclusion: The deposition of collagen reduced the expression ofα-SMA and the activation of the HSCs significantly decreased after gene transfer in the rat models with liver cirrhosis, it provides an experimental basis for treatment to liver fibrosis.