截断逆挽方对慢加急性肝衰竭大鼠肝细胞进入S期关键蛋白的影响

Study on the effect of JieDuan NiWan formula on the cell entering S phase in rats with acute-on-chronic liver failure

目的:观察截断逆挽方对慢加急性肝衰竭(ACLF)大鼠肝脏细胞进入S期关键蛋白的影响。方法:将150只Wistar大鼠随机分成3组,正常组(NC)、模型组(MC)及截断逆挽方组(JD)。采用人血清白蛋白联合D-氨基半乳糖和脂多糖急性攻击制作ACLF模型。JD组大鼠在急性攻击24 h后连续灌胃,在第5、10、15天平行取材。运用HE染色法观察大鼠肝脏组织结构,Western blot法检测转录因子E2F1的表达量,RT-PCR检测CyclinA、E和Cdc25 mRNA表达量。结果:与模型组相比,JD组大鼠E2F1的表达量明显增高,并在第10天时差异最为显著(P<0.05),CyclinA、E、Cdc25表达量均有不同程度升高;与正常组相比,JD组大鼠10 d、15 d时CyclinA mRNA表达量差异显著,JD组大鼠在10 d、15 d时Cdc25和CyclinE mRNA表达量均差异显著(P<0.05)。结论:截断逆挽方可以通过调节E2F1介导的细胞再生信号通路上的关键蛋白E2F1、CyclinA、CyclinE、Cdc25的表达,从而促使细胞进入S期,加快DNA复制进程,提高肝细胞的增殖率,发挥治疗ACLF的作用。

Objective:To observe the effect of truncated inverse pull recipe on the key proteins of the hepatocytes entering the S phase in slow adding acute liver failure(ACLF)rats.Methods:A total of 150 Wistar rats were randomly divided into 3 groups:normal group(NC),model group(MC),and truncated inverse pull group(JDNW).ACLF was modeled by human serum albumin combined with D galactosamine and lipopolysaccharide.The JDNW group received continuous gavage 24 h after the acute attack,and the samples were taken in parallel on the 5 d,10 d,and 15 d.The structures of rat liver tissues were observed by HE staining,the expression of transcription factor E2F1 was detected by Western blot,and the expression of CyclinA,E,and Cdc25 mRNA was detected by RT PCR.Results:Compared with the model group,the expression level of E2F1 in each JDNW group was significantly increased,and the difference was the most significant on the 10th day(P<0.05).The expression levels of CyclinA,E and Cdc25 were all increased to different degrees.Compared with the normal group,the expression levels of CyclinA mRNA,Cdc25,and CyclinE mRNA in the JDNW were significantly different on 10 d and 15 d(P<0.05).Conclusion:JieDuan NiWan Formula may regulate the expression of key proteins E2F1,CyclinA,CyclinE,and Cdc25 in the cell regeneration signaling pathway mediated by E2F1,so as to promote the cell to enter the S phase,accelerate the DNA replication process,improve the proliferation rate of liver cells,so as to play the role of ACLF therapy.