【作者】 郭春秀；

【导师】 谭达全；

【作者基本信息】 湖南中医药大学 ， 中医基础理论， 2014， 博士

【摘要】 目的：结合中医“治未病”思想和脾胃气机升降失常的病理,预先给予辛开苦降法中调理脾胃气机的代表方剂半夏泻心汤,观察其对水浸束缚应激导致的大鼠胃黏膜损伤、血清和胃黏膜相关细胞因子EGF、PGE2、IL-1β、IL-8、IL-10、TNF-α含量,凋亡相关物质p38MAPK、 Bcl-2、Bax、caspase-3的表达,探讨辛开苦降法防治应激性胃黏膜损伤过程中对胃黏膜上皮细胞凋亡的影响,以及发生作用的机制。方法：SD大鼠随机分为四组：空白组、模型组、中药组、西药组。采用WRS法制备大鼠应激性胃黏膜损伤模型,以半夏泻心汤为被试因素,参照Guth标准观察记录大鼠胃黏膜损伤指数(UI),观察胃黏膜组织形态学变化；采用酶免法(ELISA)检测IL-1β、IL-8、IL-10、 TNF-α、EGF、PGE2的含量；免疫组化法检测p-p38MAPK蛋白表达；RT-PCR检测Bel-2、BaxmRNA表达；Western-blot检测caspase-3蛋白表达。结果：(1)造模后,模型组大鼠胃黏膜损伤明显,损伤指数(UI)最高,与空白组有显著性差异(p<0.01),说明WRS法造模成功；中药组大鼠胃黏膜损伤指数明显降低,与模型组比较有显著性差异(p<0.01)。(2)中药组可升高WRS大鼠胃黏膜EGF、PGE2的含量,降低WRS大鼠血清IL-1β、IL-8、TNF-α的含量,升高IL-10的含量,与模型组比较有显著性差异(p<0.01)。(3)中药组可下调p-p38MAPK蛋白、上调Bcl-2mRNA表达,与模型组比较有显著性差异(p<0.01)。造模后,活化的caspase-3表达明显增加,与空白组比较有显著性差异。中药组可显著抑制caspase-3的活化,抑制胃黏膜上皮细胞发生凋亡,与模型组有显著性差异(p<0.01)。结论：(1)辛开苦降法能显著对抗应激性胃黏膜损伤,降低应激性胃黏膜损伤程度。(2)辛开苦降法能升高胃黏膜保护因子EGF、PGE2、IL-10的含量,降低胃黏膜攻击因子IL-1β、IL-8、TNF-α的含量,发挥对胃黏膜的抗炎性损伤、抗凋亡保护作用。(3)辛开苦降法可下调能引发胃黏膜上皮细胞凋亡信号转导的p-p38MAPK的表达,上调抑凋亡蛋白Bc1-2mRNA的表达,抑制细胞凋亡执行因子caspase-3的活化,从而抑制胃黏膜上皮细胞的凋亡。更多还原

【Abstract】 Objective:Combination of traditional Chinese medicine "treatment of pathological disease not" thought and the spleen and stomach qi,pre administration of pungent dispersing and bitter descending method regulating qi activity of spleen and stomach in the representative prescription of Banxia Xiexin decoction. Observe the content of stress-induced gastric mucosal injury, rat serum and gastric mucosa associated cytokines EGF、PGE2、IL-1β、IL-8、IL-10、TNF-α,and. the expression of apoptosis related substances p38MAPK、Bcl-2、 Bax、caspase-3; Investigate pungent-opening and bitter-descending method of prevention in the process of stress-induced gastric mucosal injury effect on gastric epithelial cell apoptosis, and it’s mechanism and effect.Methods:SD rats were randomly divided into four groups:blank group, model group, Chinese medicine group, western medicine group, Model of stress-induced gastric mucosal injury in rats by WRS method, Taking Banxia Xiexin decoction as subjects factors, according to the standard of Guth to Observe and record the ulcer index (UI) of gastric mucosa in rats,observe the morphological changes of gastric mucosa; Using enzyme immunoassay (ELISA) to detect the content of IL-1β、IL-8、 IL-10、TNF-α、EGF、PGE2,immunohistochemistry was used to detect the expression of p-p38MAPK protein, RT-PCR was used to To detect the expression of Bcl-2, Bax protein, Western-blot was used to To detect the expression of caspase-3.Results:(1) After modeling, injury of gastric mucosa of the rats in the model group is obvious, The Ulcer index(UI) is the highest, there were significant compared with the normal group (p<0.01), indicating a successful model by WRS method; The injury index of gastric mucosa in rats decreased significantly, there were significant compared with the model group (p<0.01),(2)After modeling, activation of Caspase-3expression increased significantly, there were significant compared with the blank group; Traditional Chinese medicine group could significantly inhibit the activation of Caspase-3, Inhibit the apoptosis of gastric epithelial cell, there were significant compared with the model group (p<0,01).(3)In Traditional Chinese medicine group, Significant up-regulation of the content of EGF, PGE2was observed in gastric mucosa of WRS rats, down-regulation of the content of IL-1β、IL-8、TNF-α and up-regulation of the content of IL-10was observed in serum of WRS rats, there were significant compared with the model group(p<0.01).(4)Traditional Chinese medicine group could down regulate p-p38MAPK expression, up-regulate the expression of Bcl-2 protein, there were significant compared with the model group (p<0.01).Conclusions:(1) Pungent dispersing and bitter descending method can significantly damage the gastric mucosa against stress, reduce the incidence of stress ulcer.(2) Pungent dispersing and bitter descending method can perform significantly inhibit apoptosis factor caspase-3activation, thereby inhibiting apoptosis of gastric epithelial cells.(3) Pungent dispersing and bitter descending method can up regulate the content of gastric mucosal protective factor EGF, PGE2, IL-10, down regulate the content of gastric mucosal attacking factor IL-1β、IL-8、TNF-α, play the anti inflammation injury, anti apoptotic protective effect on gastric mucosa.(4) Pungent dispersing and bitter descending method can down regulate the expression of p-p38MAPK which can cause gastric mucosal epithelial cell apoptosis signal transduction, up regulate the anti apoptosis protein Bcl-2; so then inhibit the epithelial cell apoptosis of gastric mucosa.更多还原