电针促进应激性溃疡小鼠胃黏膜损伤修复的实验研究

Experimental study on electric acupuncture promoting repair of gastric mucosal injury in mice with stress ulcer

目的观察电针对应激性溃疡小鼠胃黏膜损伤修复的促进作用,探讨电针的修复作用与促进修复因子释放及上调低氧诱导因子-1α(HIF-1α)表达水平的相关性。方法雄性C57BL/6小鼠随机分为对照组(Sham组)、模型组(CRS组)及模型+电针组(CRS+EA组),每组9只。除Sham组外,CRS组、CRS+EA组通过3 d冷束缚应激制备应激性溃疡模型(CRS模型),造模3 d后进行电针干预,CRS+EA组电针“足三里”穴和“梁丘”穴,时间为15 min,上、下午各1次,持续2 d。实验开始后第6天小鼠全胃取材,评估胃黏膜溃疡指数(UI)评分,实时荧光定量逆转录聚合酶链式反应(RT-qPCR)和免疫印迹(Western blot)法测定修复因子[包括三叶因子2(TFF2)、血管内皮生长因子(VEGF)、热休克蛋白70(HSP70)]以及调控因子低氧诱导因子-1α(HIF-1α)表达水平。结果①与Sham组比较,CRS组、CRS+EA组UI评分显著升高(P<0.05);与CRS组比较,CRS+EA组UI评分显著下降(P<0.05)。②与Sham组比较,CRS组VEGF、HSP70水平显著升高(P<0.05),CRS+EA组TFF2、VEGF、HSP70水平显著升高(P<0.05);与CRS组比较,CRS+EA组TFF2、VEGF、HSP70水平显著升高(P<0.05)。③与Sham组比较,CRS组HIF-1α表达差异无统计学意义(P>0.05),CRS+EA组HIF-1α表达显著升高(P<0.05);与CRS组比较,CRS+EA组HIF-1α表达显著升高(P<0.05)。结论电针可以促进修复因子(TFF2、VEGF、HSP70)的释放,促进应激性溃疡小鼠胃黏膜损伤后的修复,且电针的疗效可能与治疗后HIF-1α的表达水平显著上调相关。

Objective To observe the effect of electric acupuncture(EA)on promoting the repair of gastric mucosal injury in mice with stress ulcer,and explore the correlation between repair effect of EA and promoting repair factor release and up-regulating hypoxia-inducible factor-1α(HIF-1α)expression.Methods Male C57BL/6 mice were randomly divided into control group(Sham group),model group(CRS group)and model+EA group(CRS+EA group),with 9 mice in each group.Except for the Sham group,the stress ulcer model(CRS model)was established in the CRS and CRS+EA groups after 3 days of cold-restraint stress,and EA intervention was performed 3 d after modeling.In the CRS+EA group,EA was applied to Zusanli(ST 36)and Liangqiu(ST 34)for 15 min,once in the morning and once in the afternoon,for 2 d.On day 6 of the experiment,the whole stomachs were separated from mice and were sampled to assess the gastric mucosal ulcer index(UI)score.The mRNA expression levels of repair factors[including trefoil factor 2(TFF2),vascular endothelial growth factor(VEGF)and heat shock protein 70(HSP70)]were determined by RT-qPCR,and the protein content and mRNA expression level of HIF-1αwere determined by Western blot and RT-qPCR.Results①The CRS group and the CRS+EA group had significantly higher gastric mucosal UI score than the Sham group(P<0.05);and the CRS+EA group had significantly lower UI score than the CRS group(P<0.05).②The CRS group had significantly higher levels of VEGF and HSP70 than the Sham group(P<0.05),and the CRS+EA group showed significantly higher levels of TFF2,VEGF and HSP70 than the Sham group(P<0.05).The CRS+EA group had significantly higher levels of TFF2,VEGF and HSP70 than the CRS group(P<0.05).③There was no statistically significant difference in HIF-1αexpression between the CRS group and the Sham group(P>0.05),while the CRS+EA group had significantly higher HIF-1αexpression than the Sham group(P<0.05)and the CRS group(P<0.05).Conclusion Electric acupuncture promotes the expression of repair factors(TFF2,VEGF,HSP70)and facilitates the repair of injured gastric mucosa in mice with stress ulcer.The efficacy of EA may be associated with a significant upregulation of HIF-1αexpression after treatment.