艾灸中脘、足三里穴对急性胃粘膜损伤大鼠热休克蛋白及细胞凋亡、Smac表达的影响
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摘要
目的:艾灸预处理“中脘”、“足三里”穴观察其对急性胃黏膜损伤型大鼠胃黏膜保护作用及胃粘膜上皮细胞凋亡及热休克蛋白60(HSP60)、热休克蛋白70(HSP70)、线粒体促凋亡蛋白(Smac/Diablo)等表达的影响;探讨艾灸治疗消化系统疾病的内源性保护机制,为治疗消化系统疾病提供实验依据。
     方法:60只SD大鼠完全随机分成4组,分别为空白组、模型组、艾灸中脘、足三里组、艾灸非穴位组。艾灸预处理8天,无水乙醇灌胃,建立急性胃黏膜损伤模型。采用免疫组化法检测细胞凋亡、HSP60、HSP70、Smac阳性表达。
     结果:无水乙醇造模后,模型组UI值显著高于空白组(P<0.01),说明造模成功;艾灸中脘、足三里组UI值显著低于模型组和非穴组(P<0.01),与空白组比较也有显著性差异(P<0.01)。模型组、艾灸穴位组、艾灸非穴位组HSP60、HSP70明显高于空白组(P<0.01),说明无水乙醇灌胃后胃粘膜中HSP60和HSP70表达都增强。艾灸中脘、足三里穴位组HSP60、HSP70表达显著增强,与模型组和艾灸非穴位组比较具有显著意义(P<0.01)。模型组胃粘膜细胞凋亡指数、Smac高于空白组(P<0.05),艾灸中脘、足三里组凋亡指数和Smac显著低于模型组和艾灸非穴位组(P<0.01)。艾灸中脘、足三里抑制细胞凋亡的作用明显强于非穴位对照组(P<0.01)。
     结论:艾灸“中脘”、“足三里”穴预处理能够减轻无水乙醇对胃黏膜的损伤,诱导HSP60、HSP70的过度表达,通过抑制Smac的表达抑制细胞凋亡的发生,减少伤害性刺激,从而起到保护胃黏膜的作用。艾灸穴位点要比非穴位的作用更加明显,说明艾灸对胃黏膜保护作用具有一定的穴位特异性。
Objective:To observe the effluence on expression of apoptosis、heat shock protein60(HSP60)、heat shock protein70(HSP70)、Smac/Diablo in acute gastric mucosal lesion of rat's epithelial cells and the protection of gastric mucosa after pro-moxibustion "zhongwan" and "zusanli" point. Probe endogenous protective mechanism by moxibustion treatment of digestive diseases, and result in experiment evidence for treatment of the digestive diseases.
     Method:Sixty rats were randomly divided into four groups, such as blank control group, model group, moxibustion at "Zusanli" and "zhongwan" and moxibustion at control points groups. Gastric ulcer model was established by intragastric administration with absolute alcohol, before pro-moxibustion once daily and continuously for 8 days. At the end of the experiments, the assaying the content of HSP60、gastric mucosa, cell apoptosis and Smac by immunohistochemical method.
     Results:Absolute-alcohol model was successfully established as long as UI value in model group's was significantly higher than in blank group's. There are no statistical differences between blank group and moxibution at points group, in which UI value was significant lower than model group and moxibustion at control points group. HSP60、HSP70 values in model group、moxibustion at points group、moxibustion at control points group were significantly higher than blank group. It proved after intragastric administration with absolute alcohol, both HSP60 and HSP70 expression were enhanced in gastric mucosa. The expression of HSP60、HSP70 in moxibustion at points group of gastric mucosa decreased highly (P<0.01),compared with model group and moxibustion at control points group. The expression of apoptosis Smac/Diablo in model group was significantly increased than blank group (P<0.05), and lower in moxibustion at points group than model group and moxibustion at control points group (P<0.01). The apoptosis in moxibustion points group were inhibited stronger than in the moxibustion at control group.
     Conclusion:The pretreatment of moxibustion points group can relieve the damage of absolute alcohol to gastric mucosa. In order to protect gastric mucosa, moxibustion can induce HSP60、HSP70 to express excessively by suppressing Smac's expressing to suppress the expressing of apoptosis and harmful stimulation. The effect of moxibustion points group is more remarkable than of moxibustion at control points group, which proves the protection of gastric mucosa by moxibustion has a specificity of acupuncture.
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