氧化苦参碱联合全反式维甲酸干预大鼠肝癌变进程中细胞因子的变化

Effects of oxymatrine combined with all-trans retinoic acid on cytokines during hepatocarcinogenesis in rats

目的观察氧化苦参碱(oxymatrine,OMT)联合全反式维甲酸(all-trans retinoic acid,ATRA)抑制二乙基亚硝胺(DEN)诱导的大鼠肝癌形成效果,并探讨其作用机制。方法采用DEN诱导建立肝癌大鼠模型,造模18周,将Wistar大鼠随机分为空白组、模型组、干预组,分别在诱癌后6、12、18周处死大鼠,通过HE染色观察大鼠肝组织病理变化,检测大鼠肝功能相关指标,ELISA检测大鼠IL-6含量;采用RT-qPCR法检测大鼠let-7a、NF-κB-p65、IL-6,STAT3 mRNA表达变化;采用Western blot检测大鼠STAT3、p-STAT3的蛋白变化。结果HE染色病理学观察显示,干预组大鼠肝癌结节数明显少于模型组,肝细胞病变坏死减轻;与模型组比较,干预组大鼠血清IL-6及肝功能指标ALT、GGT、AST、ALP水平均明显降低(P<0.05);RT-qPCR检测结果:与模型组比较,干预组大鼠肝组织中let-7a mRNA表达水平明显上升,而NF-κB-p65、IL-6和STAT3 mRNA表达量均明显降低(P<0.05)。实验第18周,Western blot检测结果干预组STAT3、p-STAT3蛋白表达均明显低于模型组(P<0.05)。结论氧化苦参碱联合全反式维甲酸能够明显抑制DEN诱发的大鼠肝癌形成,在一定程度上延缓肿瘤生长。

Objective To observe the inhibitory effect of oxymatrine(OMT)combined with all-trans retinoic acid(ATRA)on diethylnitrosamine(DEN)-induced hepatocellular carcinoma in rats,and to explore its mechanism of action.Methods A rat model of hepatocellular carcinoma was established by DEN induction for 18 weeks.Wistar rats were randomly divided into control,model,and intervention groups.Rats were killed at 6,12,and 18 weeks after cancer induction.Pathological changes of the liver tissues were observed by hematoxylin and eosin(HE)staining,related indexes of rat liver function were detected,and interleukin(IL)-6 expression was detected by enzyme-linked immunosorbent assay.The mRNA expressions of let-7a,nuclear factor(NF)-κB-p65,IL-6,and signal transducer and activator of transcription(STAT)3 were detected by quantitative real-time(RT-q)PCR,while the changes of STAT3 and p-STAT3 proteins were detected by western blotting.Results The pathological examination revealed a significantly decreased number of liver cancer nodules in the intervention group compared with the model group(P<0.05),while levels of serum IL-6,alanine aminotransferase(ALT),glutamyl transpeptidase(GGT),aspartate aminotransferase(AST),and alkaline phosphatase(ALP)were significantly lower(P<0.05).RT-qPCR results showed that the expression of let-7a was significantly increased in the intervention group compared with the model group,while that of NF-κB-p65,IL-6,and STAT3 was significantly decreased(P<0.05).At the 18 th week of the experiment,the expressions of STAT3 and p-STAT3 in the intervention group were significantly lower than in the model group(P<0.05).Conclusions OMT combined with ATRA can significantly inhibit DEN-induced hepatocarcinogenesis in rats and delay the growth of tumors to some extent.