岗稔根正丁醇提取物对CCl4诱导小鼠肝纤维化中ERK/MAPK通路的影响

Effect of N-butanol Extraction of Rhodomyrtus tomentosa Root on ERK/MAPK Pathway in Liver Fibrosis Rats Induced by CCl4

目的探讨岗稔根正丁醇提取物(N-butanol extraction of Rhodomyrtus tomentosa root,N-RHT)对肝纤维化小鼠肝脏炎症的影响及机制。方法采用血小板衍生生长因子(PDGF)诱导活化的肝星状细胞(HSC)模型,并以四氯化碳橄榄油溶液[25%,CCl_(4)∶橄榄油=1∶3,2 mL/(kg·d)]腹腔注射建立小鼠肝纤维化模型。将C57BL/6小鼠分为5组:对照组、模型组、秋水仙碱(Colchicine)组、N-RHT低剂量和高剂量组(0.5 g/kg和1 g/kg)。对照组小鼠予以1.5 mL/(kg·d)纯橄榄油腹腔注射,余下各组小鼠均给予四氯化碳腹腔注射,3次/周,持续5周。苏木精-伊红、Masson和天狼星红染色观察肝脏病理变化情况;检测小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、羟脯胺酸(HYP)及PDGF的含量;Western blot法检测肝脏α-平滑肌肌动蛋白(α-SMA)以及STAT3、Akt、P44/42、P38及相应磷酸化蛋白的表达。结果细胞实验中,N-RHT可以抑制活化的肝星状细胞的增殖,减少肝星状细胞中α-SMA的表达(均P<0.01),下调肝星状细胞中磷酸化P44/42蛋白(p-P44/42)的表达(P<0.01)。动物实验中,与模型组比较,N-RHT可以改善CCl_(4)导致的肝纤维化病理损伤,减少肝组织中α-SMA的表达(P<0.05),增加血清中SOD、GSH-Px的含量(均P<0.05),降低血清中ALT、AST、PDGF、HYP以及MDA的含量(均P<0.05)。结论N-RHT具有改善CCl_(4)诱导的小鼠肝纤维化的作用,其机制可能与抑制ERK/MAPK信号通路的激活有关。

Objective To observe the effect of N-butanol extraction of Rhodomyrtus tomentosa root(N-RHT)on liver in-flammation in carbon tetrachloride(CCl_(4))-induced liver fibrosis mice,and to explore its mechanism.Methods PDGF was used to induce activated hepatic stellate cell(HSC)model,and carbon tetrachloride olive oil solution[25%,CCl_(4):olive oil=1:3,2 mL/(kg·d)]was injected intraperitoneally to establish liver fibrosis model in mice.C57BL/6 mice were divided into 5 groups:control group,model group,Colchicine group,N-RHT low dose and high-dose groups(0.5 g/kg and 1 g/kg).The 1.5 mL/(kg.d)olive oil was injected intraperitoneally in control group,and mice in the remaining groups were injected intraperitoneally with carbon tetrachloride 3 times/week for 5 weeks.HE,Masson and Sirius red staining were used to observe pathological dam-age.Serum levels of ALT,AST,SOD,MDA,GSH-Px,HYP and PDGF in mice were detected.The expression levels of ar smooth muscle actin(a SMA),STAT3,Akt,P44/42,P38 and corresponding phosphorylated proteins in liver were detected by Western blotting.Results In cell experiments,N-RHT significantly inhibited proliferation of activated hepatic stellate cells,reduced a-SMA expression(both P<0.01),and downregulated the expression of p P44/42 protein in hepatic stellate cells(P<0.01).In the animal experiment,compared with the model group,the pathological damage of liver fibrosis induced by CCl_(4)was obviously improved in N-RHT group,and the expression of ar SMA in liver tissues was significantly decreased(both P<0.05).The con-tents of serum SOD and GSH-Px were significantly increased(both P<0.05).The contents of serum ALT,AST,PDGF,HYP and MDA were significantly decreased(all P<0.05).Conclusion N-RHT can improve CCI4-induced liver fibrosis in mice by inhibiting activation of ERK/MAPK signaling pathway.