siRNA沉默c-jun基因对肝硬化大鼠肝切除术后肝再生的影响及其机制

Influence and mechanism of c-jun gene silenced by siRNA on liver regeneration in liver cirrhotic rats after hepatectomy

目的探讨si RNA沉默c-jun基因对肝硬化大鼠肝切除术后肝再生的影响及其机制。方法建立肝硬化大鼠模型,采用随机数字表法将50只肝硬化大鼠随机分为治疗组和对照组,每组25只。治疗组行肝大部分切除术的同时门静脉内注射慢病毒包装的Pcmv6-AC-GFP/c-jun-si RNA质粒。对照组行肝大部分切除术的同时门静脉内注射PBS。两组大鼠于术后14 d检测门静脉压力、血清ALT及肝重量体重比,采用Western blot法检测肝脏c-jun和增殖细胞核抗原(PCNA)蛋白相对表达量。两组间的数据比较采用t检验。结果治疗组术后平均门静脉压力为(1.18±0.11)k Pa,明显小于对照组的(1.67±0.24)k Pa(t=-26.74,P<0.05)。治疗组术后ALT为(43±5)U/L,明显低于对照组的(257±14)U/L(t=-31.11,P<0.05)。治疗组术后肝重量体重比为(6.94±0.31)%,明显大于对照组的(2.76±0.14)%(t=23.57,P<0.05)。治疗组术后c-jun和PCNA蛋白相对表达量分别为0.143±0.014、0.195±0.027,对照组相应为0.742±0.057、0.029±0.003,差异有统计学意义(t=-37.17,14.86;P<0.05)。结论慢病毒负载的c-jun-si RNA可能通过下调肝细胞c-jun,上调PCNA蛋白表达来促进肝再生,改善肝硬化大鼠肝切除术后肝功能。

ObjectiveTo investigate the influence and mechanism of c-jun gene silenced by siRNA on liver regeneration in liver cirrhotic rats after hepatectomy.MethodsLiver cirrhotic rat models were established and 50 liver cirrhotic rats were randomized into the treatment group and the control group by random number table method with 25 rats in each group. The rats in the treatment group underwent major hepatectomy and were administered with lentivirus carrying plasmid Pcmv6-AC-GFP/ c-jun-siRNA by intraportal injection, while the rats in the control group underwent major hepatectomy and were administered with phosphate buffer saline （PBS） by intraportal injection. The portal pressure, serum ALT and liver weight/body weight of both groups were tested 14 d after surgery. The relative expression of c-jun in the liver and proliferating cell nuclear antigen （PCNA） protein was detected by Western blot. The data between the treatment group and the control group were compared usingt test.ResultsThe average portal pressure of the treatment group after surgery was （1.18±0.11） kPa, which was signiifcantly lower than （1.67±0.24） kPa of the control group （t=-26.74,P〈0.05）. ALT of the treatment group after surgery was （43±5） U/L, which was signiifcantly lower than （257±14） U/L of the control group （t=-31.11, P〈0.05）. Liver weight/body weight of the treatment group after surgery was （6.94±0.31）%, which was signiifcantly greater than （2.76±0.14）% of the control group （t=23.57,P〈0.05）. The relative expression of c-jun and PCNA protein of the treatment group after surgery was respectively 0.143±0.014 and 0.195±0.027, and those of the control group was respectively 0.742±0.057 and 0.029±0.003, where signiifcant difference was observed （t=-37.17, 14.86;P〈0.05）.ConclusionLentivirus carrying c-jun-siRNA may promote liver regeneration and improve live function in cirrhotic rats after hepatectomy by down-regulating the c-jun in hepatocyte and up-regulating the PCNA protein.