扶正化瘀方对大鼠肝再生过程中cyclinD1影响的实验研究

Experiment study of the effect of Fuzhenghuayu decoction on cyclinD1 during liver regeneration in rats

目的探讨扶正化瘀方对大鼠肝大部切除后72h肝再生过程中cyclinD1的表达和肝再生速度的影响。方法将 24只正常雄性Wistar大鼠随机分为4组,分别为假手术组(对照组)、模型(PH)组、PH+扶正化瘀组和PH+促肝细胞生长素(pHGF)组,每组6只。PH组、PH+扶正化瘀组和PH+pHGF组大鼠切除肝脏左叶和中叶(约占肝脏总重量的70%),对照组仅作假手术处理。扶正化瘀组和pHGF组于实验前3d至实验结束,分别给予扶正化瘀方药液灌胃1ml/100g和腹腔注射pHGF1ml/100g,1次/d。术后72h处死大鼠,计算4组肝再生速度、有丝分裂指数,免疫组织化学染色和荧光定量PCR检测肝组织cyclinD1的表达。结果与假手术组比较,PH组、PH+扶正化瘀组和PH+pHGF组大鼠有丝分裂指数、肝组织cyclinD1 mRNA和肝细胞核cyclinD1阳性率均显著升高(P<0.05);与PH组比较,PH+扶正化瘀组和PH+pHGF组大鼠肝再生速度、有丝分裂指数、肝组织cyclinD1 mRNA和肝细胞核cyclinD1阳性率均显著升高(P<0.05),而PH+扶正化瘀组和PH+pHGF组差异无统计学意义(P>0.05)。结论扶正化瘀方可上调正常大鼠肝大部切除术后72h时肝细胞cyclinD1的表达,从而促进肝再生。

Objective To investigate the effect of Fuzhenghuayu decoction on cyclinD1 expression and liver regeneration rate during liver regeneration in rats 72 hours after partial hepatectomy. Methods Twenty-four male Wistar rats were randomized into 4 groups ： sham-operation group （ control group）, partial hepatectomy group （ PH group）, partial hepatectomy ＋ Fuzhenghuayu decoction group （ PH ＋ Fuzheng group）, partial hepateetomy ＋ pHGF group （ PH ＋ pHGF group）, with 6 rats ineach group. The rats in PH group, PH ＋ Fuzheng group and PH ＋ pHGF group were hepatectomized （ left lobe and median lobe, about 70% of liver total weight）, however, the rats in sham-operation group were performed with sham operation only. The rats in PH ＋ Fuzheng group and in PH ＋ pHGF group were given Fuzhenghuayu decoction lml/100g by gavage and intraperitoneal injection with pHGF Iml/ 100 g,once a day,from 3 days before the experiment to end of the experiment. The rats were sacrificed 72 hours after the operation, and liver regeneration rate and mitotic indexes were calculated, and the expression levels of cyclinD1 were detected by immunohistostaining and fluorescent quantitation PCR. Results As compared with those in sham-operation group, the mitotic indexes, expression levels of cyclin D1 mRNA of liver tissues and positive rate of cyclin D1 of hepatic nucleus in PH group, PH ＋ Fuzheng group, PH ＋ pHGF group were significantly increased （ P 〈 0. 05 ） ; as compared with those in PH group, the liver regeneration rate, mitotic indexes, expression levels of cyclin D1 mRNA of liver tissues and positive rate of cyclin D1 of hepatic nucleus in PH ＋ Fuzheng group and PH ＋ pHGF group were significantly increased （ P 〈 0. 05 ）, however, there were no significant differences in these parameters between PH ＋ Fuzheng group and PH ＋ pHGF group （ P 〈 0. 05 ）. Conclusion Fuzhenghuayu decoction can up-regulate the expression of cyclinD1 of hepatocytes in rats 72 hours after partial hepatectomy, as a result, which can promote liver regeneration.