扁蒴藤素调节ROS/ASK1/JNK信号通路对二乙基亚硝胺诱导大鼠肝癌的抑制作用

Inhibitory effect of pristimerin on diethylnitrosamine-induced hepatocellular carcinoma in rats by regulating ROS/ASK1/JNK signal pathway

目的探讨扁蒴藤素(Pris)通过调节ROS/ASK1/JNK信号通路对二乙基亚硝胺(DEN)诱导大鼠肝细胞癌(HCC)的影响。方法随机取6只SD大鼠作为对照组,其余大鼠采用注射DEN的方式构建HCC大鼠模型。将造模成功的大鼠随机平分为HCC组、Pris组(0.8 mg/kg Pris)、Vaccarin组(100 mg/kg ROS/ASK1/JNK信号通路抑制剂Vaccarin)、Pris+Vaccarin组(0.8 mg/kg Pris+100 mg/kg Vaccarin),连续注射1周,每组均6只大鼠。HCC组和对照组注射等量生理盐水。测量体质量、肝质量以及肝脏体质量比;ELISA法检测肝功能、炎性因子、抗氧化指标、ROS水平;HE染色检测肝脏病理变化;Western blot检测凋亡标志物(Bax、Bcl-2和cleaved-Caspase-3)以及ROS/ASK1/JNK信号通路蛋白表达。结果对照组大鼠表现出正常的肝脏组织结构;HCC组可以观察到部分肝细胞坏死,出现局灶性结节性增生现象,HCC组较对照组体质量、Bax、cleaved-Caspase-3、ROS、p-ASK1/ASK1、p-JNK/JNK水平显著下降(P<0.05),肝脏质量、肝脏体质量比、ALT、AST、ALP、LDH含量、IL-6、TNF-α、CCL-2含量、SOD、GR、GPx、CAT含量、Bcl-2蛋白水平显著增加(P<0.05);Pris组改善了肝细胞坏死以及局灶性结节性增生现象,Pris组较HCC组体质量以及Bax、cleaved-Caspase-3、ROS、p-ASK1/ASK1、p-JNK/JNK水平显著升高(P<0.05),肝脏质量、肝脏体质量比、ALT、AST、ALP、LDH含量、IL-6、TNF-α、CCL-2含量、SOD、GR、GPx、CAT含量、Bcl-2蛋白水平显著降低(P<0.05),而Vaccarin组趋势相反;Vaccarin逆转了Pris对HCC大鼠的抗癌效果。结论Pris可能通过激活ROS/ASK1/JNK信号通路对HCC大鼠起到一定的抑癌作用。

Objective To investigate the influence of pristimerin(Pris)on rat hepatocellular carcinoma(HCC)induced by diethylnitrosamine(DEN)by regulating ROS/ASK1/JNK signal pathway.Methods Six SD rats were randomly selected as the control group,and the remaining rats were injected with DEN to construct the rat model of HCC.The rats successfully modeled were randomly grouped into HCC group,Pris group(0.8 mg/kg Pris),Vaccarin group(100 mg/kg ROS/ASK1/JNK signal pathway inhibitor Vaccarin),and Pris+Vaccarin group(0.8 mg/kg Pris+100 mg/kg Vaccarin),with 6 rats in each group,and were injected continuously for one week.HCC group and control group were injected with the same amount of normal saline.Body weight,liver weight and liver weight ratio were measured;liver function,inflammatory factors,antioxidant index and ROS level were detected by ELISA;HE staining was applied to detect the pathological changes of liver;western blot was applied to detect the expression of apoptotic markers(Bax,Bcl-2 and cleaved-Caspase-3)and ROS/ASK1/JNK signal pathway protein.Results The rats in the control group showed normal liver tissue structure.In HCC group,necrosis of some hepatocytes and focal nodular hyperplasia were observed,body weight,Bax,cleaved-Caspase-3,ROS,p-ASK1/ASK1,and p-JNK/JNK level in HCC group were obviously lower than those in the control group(P<0.05),but liver weight,liver weight ratio,ALT,AST,ALP,LDH contents,IL-6,TNF-α,CCL-2 contents,SOD,GR,GPx,CAT contents,and Bcl-2 protein level were obviously higher(P<0.05).Hepatocyte necrosis and focal nodular hyperplasia in Pris group were improved,and body weight,Bax,cleaved-Caspase-3,ROS,p-ASK1/ASK1,and p-JNK/JNK level in Pris group were obviously higher than those in HCC group(P<0.05);liver weight,liver weight ratio,ALT,AST,ALP,LDH contents,IL-6,TNF-α,CCL-2 contents,SOD,GR,GPx,CAT contents,and Bcl-2 protein level were obviously lower(P<0.05).The opposite trend was observed in Vaccarin group;Vaccarin reversed the anticancer effect of Pris on HCC rats.Conclusion Pris may play a certain role in inhibiting HCC rats by activating ROS/ASK1/JNK signal pathway.