蒿汤对酒精性肝纤维化大鼠肝功能和pERK/eIF2a信号通路的影响

Effect of haowu decoction on liver function and pERK/eIF2a signaling pathway in rats with alcoholic liver fibrosis

目的:探究蒿汤对酒精性肝纤维化大鼠肝功能的改善及磷酸化细胞外调节蛋白激酶/真核翻译起始因子2α(pERK/eIF2a)信号通路的影响。方法:将40只健康的雄性SD大鼠随机分成4组:正常组、假手术组、模型组及蒿汤组,每组10只。采用“酒精-玉米油-吡唑”灌胃联合12周的高脂饮食构建酒精性肝纤维化模型,造模成功后,正常组不做处理,假手术组给予生理盐水灌胃,模型组和蒿汤组于相同时间给予等量蒿汤灌胃治疗。连续治疗12周后检测各组血清羟脯胺酸(HYP)、透明质酸酶(HA)、层黏连蛋白(LN)、Ⅳ型胶原(CⅣ)、Ⅲ型前胶原(PⅢNP)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ-谷氨酰转肽酶(γ-TG)、总胆汁酸(TBA)、总胆红素(TB)、白蛋白(ALB)、总胆固醇(CHOL)、甘油三酯(TG)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平,苏木精-伊红染色法(HE)观察大鼠肝脏纤维化程度,酶联免疫吸附试验(ELISA)检测肝脏组织中pERK/eIF2a信号通路的表达水平。结果:与正常组对比,假手术组血清HYP、HA、LN、CIV、PⅢNP、AST、ALT、γ-GT、ALP、TBL、TB水平无明显改变(P>0.05),模型组显著升高(P<0.01);给予蒿汤干预治疗后,与模型组相比,大鼠血清HYP、HA、LN、CIV、PⅢNP、AST、ALT、γ-GT、ALP、TBL、TB水平显著降低(P<0.01)。血清白蛋白、脂代谢及氧化损伤指标显示,与正常组对比,假手术组血清ALB、CHOL、TG、GSH、SOD、MDA水平无明显改变(P>0.01),模型组血清ALB、GSH、SOD水平显著降低(P<0.01),CHOL、TG、MDA水平显著升高(P<0.01);给予蒿汤干预治疗后,与模型组相比,大鼠血清ALB、GSH、SOD水平显著升高(P<0.01),CHOL、TG、MDA水平显著降低(P<0.01)。HE染色结果显示,与对照组比较显示假手术组肝脏切片形态学正常,而模型组肝脏切片可见明显空泡状改变,给予蒿汤干预治疗组肝脏切片明显好转。ELISA结果显示,与正常组对比,假手术组肝脏组织中pERK和eIF2a水平无明显改变(P>0.05),模型组肝脏组织中pERK和eIF2a水平显著升高(P<0.01);给予蒿汤干预治疗后,与模型组相比,大鼠肝脏组织中pERK和eIF2a水平显著降低(P<0.01)。结论:蒿汤可以有效阻碍酒精性肝纤维化大鼠的肝脏纤维化程度,降低肝纤维化指标,改善肝胆功能。发挥这一作用可能与抑制pERK/eIF2a信号通路有关。

Objective:To explore the improvement of liver function and the effect of pERK/eIF2a signaling pathway in rats with alcoholic liver fibrosis.Methods:Forty healthy male SD rats were randomly divided into 4 groups:normal group,sham operation group,model group and Artemisia group,with 10 rats in each group.Alcoholic liver fibrosis model was established by"alcohol-corn oil-pyrazole"and 12-week high-fat diet.After successful modeling,the normal group was not treated,and the sham operation group was given saline.And the Artemisia decoction group were given the same amount of Artemisia decoction at the same time.After 12 weeks of continuous treatment,serum HYP,HA,LN,CIV,PIIINP,ALT,AST,ALP,γ-TG,TBA,TB,ALB,CHOL,TG,GSH,SOD and MDA levels were detected in each group.The degree of liver fibrosis was observed in rats by HE staining,and the expression level of pERK/eIF2a signaling pathway in liver tissue was observed by ELISA.Results:Compared with the normal group,the levels of serum HYP,HA,LN,CIV,PIIINP,AST,ALT,γ-GT,ALP,TBL and TB in the sham operation group did not change significantly(P>0.05),and the rats in the model group significantly increased.High(P<0.01);serum and HYP,HA,LN,CIV,PIIINP,AST,ALT,γ-GT,ALP,TBL,and TB levels were significantly lower in rats compared with the model group.(P<0.01).Serum albumin,lipid metabolism and oxidative damage indicators showed that there was no significant change in serum ALB,CHOL,TG,GSH,SOD and MDA levels in the sham-operated group compared with the normal group(P>0.01).The levels of GSH and SOD were significantly decreased(P<0.01),and the levels of CHOL,TG and MDA were significantly increased(P<0.01).After treatment with Artemisia sinensis,serum ALB,GSH and SOD were compared with the model group.The level was significantly increased(P<0.01),and the levels of CHOL,TG and MDA were significantly decreased(P<0.01).The results of HE staining showed that compared with the control group,the morphology of the liver sections of the sham-operated rats was normal,while the liver sections of the model group showed obvious vacuolization changes.The liver sections of the rats treated with Artemisia sinensis were significantly improved.The results of ELISA showed that there was no significant change in the levels of pERK and eIF2a in the liver tissue of the sham-operated rats compared with the normal group(P>0.05).The levels of pERK and eIF2a in the liver tissue of the model group were significantly increased(P<0.01).After treatment with Artemisia sinensis,the levels of pERK and eIF2a in rat liver tissues were significantly lower than those in the model group(P<0.01).Conclusion:Artemisia decoction can effectively block the degree of liver fibrosis in rats with alcoholic liver fibrosis,reduce liver fibrosis index,and improve hepatobiliary function.This effect may be related to inhibition of the pERK/eIF2a signaling pathway.