针刺对四氯化碳致肝纤维化大鼠血小板衍生生长因子信号通路的影响

Effects of Acupuncture Intervention on Hepatic Platelet-derived Growth Factor Signaling Pathway in CCl_4-induced Hepatic Fibrosis Rats

目的:观察针刺对肝纤维化模型大鼠血小板衍生生长因子(PDGF)信号通路蛋白和mRNA表达的影响,探讨针刺抗肝纤维化的可能机制。方法:将46只SD大鼠分为空白组10只,模型组、非穴组、针刺组,每组12只。采用50%四氯化碳(CCl4)和橄榄油腹腔注射复制肝纤维化模型。针刺"太冲""期门""肝俞",电针"足三里"。采用Western blot和Real-time PCR方法检测各组大鼠PDGF信号通路蛋白及其mRNA表达。结果:与空白组相比较,模型组大鼠血清PDGF含量及肝脏PDGF信号通路相关蛋白、mRNA表达明显升高(P<0.01,P<0.05);与模型组相比,针刺组则能够抑制PDGF-βR及其下游细胞外调节蛋白激酶(ERK)蛋白与mRNA的表达(P<0.01,P<0.05);而针刺对Jun细胞核激酶(JNK)、P 38蛋白的表达无明显调节作用(P>0.05)。非穴组与模型组相比,各指标蛋白与mRNA的表达差异均无统计学意义(P>0.05)。结论:针刺通过抑制CCl4致肝纤维化大鼠的PDGF信号通路,减少胶原沉积,促进细胞外基质的降解,从而起到治疗肝纤维化的作用。

Objective To observe the effect of acupuncture stimulation of ＂Taichong＂（LR 3）,＂Qimen＂（LR 14）,etc.on hepatic platelet-derived growth factor（PDGF） signal pathway activity at the protein and mRNA levels in hepatic fibrosis rats.Methods Forty-six SD rats were randomly divided into control（10 rats）,model（12 rats）,acupuncture（12 rats） and non-acupoint（12 rats） groups.Hepatic fibrosis model was established by intraperitoneal injection of mixture solution of 50% CCl4 and olive oil [1∶1,3 times on the 1st week（W）,twice/W thereafter for 5 more weeks].During modeling,acupuncture stimulation of ＂Taichong＂（LR 3）,＂Qimen＂（LR 14）,＂Ganshu＂（BL 18） and ＂Zusanli＂（ST 36） was conducted simultaneously.At the end of the experiments,all the rats were sacrificed for collecting their liver and blood samples,followed by separation of the hepatic stellate cells（HSCs）.ELISA,Western blot and Real-time quantitative PCR techniques were used to detect the content of serum PDGF and expression levels of PDGF-β receptor（PDGF-β R）,extracellular signal-regulated kinase（ERK1/2）,c-jun N-terminal kinase（JNK） and P 38 genes and proteins of HSCs,respectively.Results Compared to the control group,serum PDGF content,and expression levels of PDGF-β R mRNA and protein,ERK mRNA and protein and P 38 protein of HSCs in the model group were upregulated significantly（P0.01,P0.05）.In comparison with the model group,serum PDGF content,and the expression levels of PDGF-β R mRNA and protein,ERK mRNA and protein of HSCs in the acupuncture group were down-regulated appa-rently（P0.05,P0.01）.No significant differences were found between the acupuncture and non-acupoint groups in serum PDGF content and between the model group and non-acupoint group in the expression levels of PDGF-β R mRNA and protein,ERK mRNA and protein,JNK protein and P 38 protein of HSCs,as well as between the model group and acupuncture group in the expression levels of JNK protein and P 38 protein of HSCs（P0.05）.Conclusion Acupuncture intervention can effectively down-regulate serum PDGF content,and expression levels of PDGF-β R mRNA and protein,ERK mRNA and protein of HSCs in liver fibrosis rats,which may contribute to its effect in improving liver fibrosis through down-regulating PDGF signal pathway activity.