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目的:通过槲皮素灌胃阻断HSP70表达后,观察艾灸预处理对大鼠急性黏膜损伤指数及细胞凋亡影响,以进一步探明艾灸的细胞保护作用及其与HSP70的关系。方法:40只大鼠随机分为:A捆绑对照组,B模型组,C艾灸+模型组,D槲皮素+艾灸+模型组,E槲皮素+模型组。D、E组每日采用槲皮素灌胃。C、D组每日艾灸足三里、梁门穴,以上处理共8天。第9天B、C、D、E组均采用无水乙醇灌胃制作大鼠急性胃黏膜损伤模型。实验结束后wester-blot方法检测胃黏膜HSP70蛋白表达;TUNEL法检测胃黏膜细胞凋亡指数(AI)。结果:①与A组比较,模型组大鼠胃黏膜细胞凋亡指数明显增高(P<0.05),胃黏膜HSP70表达升高(P<0.01)。②与B组相比,C组大鼠细胞凋亡指数(AI)明显降低(P<0.05),HSP70表达显著升高(P<0.01);D组大鼠AI值明显增加(P<0.01),胃黏膜HSP70表达未见上调。③与C相比,D组大鼠AI值升高明显(P<0.01),黏膜HSP70表达明显下调。结论:艾灸预处理上调胃黏膜HSP70表达,急性病胃黏膜损伤所致的细胞调亡被抑制。槲皮素阻断HSP70的合成后,艾灸预处理抑制急性胃黏膜损伤细胞凋亡的作用被取消,再一次证明艾灸预处理对胃黏膜的保护作用,确与艾灸上调HSP70表达有关。
OBJECTIVE: To observe the effect of moxibustion pretreatment on acute mucosal injury index and apoptosis in rats after intragastric administration of quercetin to block the expression of HSP70, in order to further explore the cytoprotective effect of moxibustion and its relationship with HSP70. Methods: Forty rats were randomly divided into: A control group, B model group, C moxibustion + model group, D quercetin + moxibustion + model group, and E quercetin + model group. Groups D and E received quercetin daily. Group C and D daily moxibustion Zusanli and Liangmen points were treated for 8 days. On the 9th day, rats in groups B, C, D, and E were intragastrically administered with anhydrous ethanol to produce acute gastric mucosal injury models. After the end of the experiment, we detected the expression of HSP70 protein in gastric mucosa by western-blot method, and detected the apoptosis index (AI) in gastric mucosa by TUNEL method. Results: 1 Compared with group A, the apoptotic index of gastric mucosa in the model group was significantly higher (P<0.05), and the expression of HSP70 in gastric mucosa was increased (P<0.01). 2 Compared with group B, the apoptosis index (AI) in group C was significantly decreased (P<0.05), and the expression of HSP70 was significantly increased (P<0.01); the AI value in group D was significantly increased (P<0.01). There was no up-regulation of HSP70 expression in gastric mucosa. 3 Compared with C, the AI value in group D increased significantly (P<0.01), and the expression of HSP70 in mucosa was significantly down-regulated. Conclusion: Moxibustion pretreatment up-regulates the expression of HSP70 in gastric mucosa, and apoptosis induced by acute gastric mucosal injury is inhibited. After quercetin blocked the synthesis of HSP70, the effect of moxibustion pretreatment on inhibiting apoptosis in acute gastric mucosal injury was abolished. Once again, the protective effect of moxibustion pretreatment on gastric mucosa was proved to be related to the up-regulation of HSP70 expression by moxibustion.