摘要
目的:观察"截断逆挽方"对慢加急性肝衰竭(Acute-on-Chronic Liver Failure,ACLF)大鼠肝组织中周期蛋白D1(Cyclin D1)、周期蛋白A(Cyclin A)、周期蛋白依赖激酶4(CDK4)、转录因子DP1表达的影响。探讨该方预防性干预ACLF大鼠肝细胞增殖途径的作用机制。方法:SPF级Wistar雄性大鼠150只,随机分为正常对照组、模型组和截断逆挽方组,给予人血清白蛋白免疫诱导大鼠形成肝硬化模型,再联合D-氨基半乳糖(D-Gal N)与脂多糖(LPS)急性攻击,建立ACLF大鼠模型。截断逆挽方组在急性攻击前给予截断逆挽方连续灌胃3天。各组大鼠分别在急性攻击后4、8、12 h取材。蛋白质印迹法(Western Blot)检测DP1蛋白表达量,实时荧光定量qRT(Quantitative Real-Time,qRT)PCR法检测肝组织中CyclinD1、CyclinA、CDK4 mRNA的含量。结果:与正常组相比,模型4 h组CyclinD1、CyclinA、CDK4及DP1表达量升高(P<0.05);8 h、12 h随着时间的推移表达量逐渐降低。与对应时间点模型组相比,截断逆挽方4 h组CyclinD1、CyclinA、CDK4及DP1表达量无明显差异,8h CyclinD1、CyclinA、CDK4及DP1表达量有升高趋势,12 h组CyclinD1、CyclinA、CDK4及DP1表达量较模型组明显升高(P<0.01)。结论:截断逆挽方可能通过调节E2F1信号通路中CyclinD1、CyclinA、CDK4及DP1的表达量,促进了肝细胞的代偿性增殖,从而改善肝细胞的大面积坏死,抑制肝衰竭的发展。
Objective： To observe the effect of Jieduan Niwan（JDNW） Formula on the expressions of CyclinD1,CyclinA, CDK4, and transcription factor DP1 in acute - on - chronic liver failure（ACLF）rats, and to discussthe hypatocyte proliferation pathway and the action mechanism of the formula for ACLF rats. Methods： 150male Wistar rats were randomly divided into the blank control group, the model group, and JDNW group. Human serum albumin was used to induce liver cirrhosis, and acute attack of D - GalN and LPS were applied afterwards to establish ACLF models. Rats in JDNW group were orally given JDNW Formula for 3 days beforethe acute attack. The samples were collected 4h, 8h, and 12h after the acute attack, expressions of DP1, CyclinD1, CyclinA, CDK4mRNA were detected with Western blot method and qRT - PCR analysis respectively.Results： Compared to the blank control group, the expressions of CyclinD1, CyclinA, CDK4, and DP1 weresignificant increased 4 h after the acute attack in the model group（P 〈 0. 05）, and they were gradually de￣creased with time went by. Compared to the model group, there were no significant differences in the levels ofCyclinD1, CyclinA, CDK4 and DP1 in Jieduan Niwan group at 4 h, and they showed increasing tendency at8 h, and reached the peak at 12 h（P 〈 0. 01）. Conclusion： Jieduan Niwan Formula can promote the compen￣satory proliferation of hepatocytes and restrain hepatic failure by adjusting the expression of CyclinD1, CyclinA, CDK4 and DP1 of E2F1 signaling pathway.
作者
郝玉林
杨文龙
侯伟欣
田甜
张秋云
高连印
杜宇琼
HAO Yulin;YANG Wenlong;HOU Weixin;TIAN Tian;ZHANG Qiuyun;GAO Lianyin;DU Yuqiong(School of TCM,Capital Medical University,Beijing Key Laboratory ofTCM Collateral Disease Research,Beijing 100069,China)
出处
《中医药信息》
2018年第3期12-17,共6页
Information on Traditional Chinese Medicine
基金
国家自然科学基金项目(No.81573767)
关键词
慢加急性肝衰竭
截断逆挽方
细胞增殖
E2F1信号通路
Acute - on - chronic liver failure
Jieduan Niwan Formula
Cell proliferation
E2F1 signaling pathway
