目的:选择性地切断大鼠胃迷走神经通路,并应用组织形态学、生化及分子生物学等方法,观察艾灸预处理对胃黏膜损伤大鼠的内源性物质白细胞介素-1β(IL-1β)和表皮生长因子(EGF)、肿瘤坏死因子(TNF-α)含量的影响,以及热休克蛋白70的表达,进一步探讨艾灸预处理对胃黏膜损伤的保护作用机制和迷走神经通路之间的关系。方法:将SD大鼠随机分为四组,即A正常对照组;B模型组;C艾灸+模型组;D艾灸+模型+迷走神经切断组。手术组应严格按照要求先进行迷走神经切断,然后采用无水乙醇灌胃法对相应各组制备大鼠急性胃黏膜损伤模型,同时对各组进行艾灸处理,取胃组织后按Guth法计算胃黏膜损伤指数(UI)和光镜下观察胃黏膜组织形态学变化;最后采用酶联免疫吸附法检测胃黏膜组织保护相关内源性物质IL-1β、EGF、TNF-α的含量和蛋白质印迹法(Western-blot)检测血清中HSP70的表达。结果:与A组比较,B组UI、血清IL-1β含量明显升高(P〈0.01)、EGF含量下降(P〈0.01)、TNF-α含量明显升高(P〈0.01)、胃黏膜组织浸润明显、HSP70稍升高;与B组和D组比较,C组UI、血清IL-1β含量明显降低(P〈0.01)、TNF-α含量明显降低(P〈0.01)、EGF含量升高(P〈0.01)、胃黏膜组织形态较为完好、HSP70表达上调(P〈0.05)。结论:艾灸组能够明显改善大鼠胃黏膜损伤组织形态和降低胃黏膜损伤指数,同时也能上调大鼠胃黏膜当中HSP70的表达,而这一修复作用在一定程度上受到了迷走神经的调节,因而,迷走神经通路是艾灸对胃黏膜损伤修复和保护因子表达的重要调节途径。
Objective :To study the effects of moxibustion preconditioning on endogenous substance - 1 ( IL - 1 ) ,tumor necrosis factor (EGF) ,tumor necrosis factor (TNF -α), and expression of heat shock protein (HSP70) in rats with gastric mucosal injury and to further explore the relationship between the protective effect of moxibustion preconditioning on gastric mucosal injury and the vagus nerve pathway by selectively cutting off the vagus nerve pathway in the rat and using the method of tissue morpholo- gy, biochemistry and molecular biology. Methods : The SD rats were randomly divided into four groups, namely A normal control group, B model group, C moxibustion + model group, D moxibustion + model + vagus nerve transection group. The rats in the opera- tion group were treated with the method of absolute ethanol, and then the rats were treated with UI method, and the gastric mucosa tissue was detected by Guth method. The expression of IL - 1, TNF - α, EGF and Western - blot was detected by assay. The expression of heat shock protein 70 ( HSP - 70 ) was detected by enzyme - linked immunosorbent assay (ELISA). Results : Compared with A group, UI, serum IL - 1, B, EGF, TNF - α, P 〈 0.01, HSP70, P 〈 0.01, UI, IL - 1, B, P 〈 0.01, P 〈 0.01, P 〈 0.01, EGF,HSPTO and D,P 〈0.05 and C were significantly lower (TNF -α). Conclusion:Moxibustion group can significantly improve the gastric mucosal damage tissue morphology and reduce the gastric mucosa damage index, but also can increase the expression of HSP- 70 in gastric mucosa of rats, and this repair function to a certain extent by the regulation of vagus nerve, and so on.