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[目的]实时定量PCR检测冬胃颗粒对寒邪客胃型胃溃疡大鼠胃黏膜细胞ECE-1表达的影响,在基因水平探讨冬胃颗粒防治寒性胃溃疡的作用机制。[方法]采用乙酸注射法制备大鼠胃溃疡模型,随机分为正常对照组、溃疡模型组、寒性溃疡组、西药治疗组、中药治疗组。寒性溃疡组及治疗组制成溃疡模型后采用寒冷因素刺激复制寒邪客胃型动物模型,前3天用0℃冰水浸泡,每次10min,后3天用0℃冰水灌胃,治疗组同时给予药物灌胃。测定大鼠胃黏膜溃疡指数(UI)及溃疡抑制率,应用ELISA法检测胃黏膜ET-1含量及实时定量PCR法检测ECE-1mRNA转录表达水平。[结果]中药治疗组溃疡指数明显低于溃疡模型组与寒性溃疡组(P<0.01),及西药治疗组(P<0.05),其溃疡抑制率为54.22%,明显高于西药治疗组的45.40%。溃疡模型组及寒性溃疡组大鼠胃黏膜ET-1水平与正常对照组比较明显增加(P<0.01)。中药治疗组胃黏膜ET-1水平明显降低,且接近正常对照组,与溃疡模型组及寒性溃疡组比较差异均有统计学意义(P<0.05,P<0.01)。相关性分析显示:胃黏膜ET-1水平与UI呈显著正相关(r=0.89,P<0.01)。溃疡模型组及寒性溃疡模型组大鼠胃黏膜ECE-1mRNA的转录表达水平明显增加(P<0.05,P<0.01),药物治疗可下调胃黏膜ECE-1mRNA的转录表达水平,冬胃颗粒治疗组优于西药治疗组(P<0.05),与胃黏膜组织中ET-1水平变化趋势一致,二者呈显著正相关(r=0.91,P<0.01)。[结论]寒性刺激可增加乙酸性胃溃疡大鼠胃黏膜损害,胃黏膜ET-1水平及ECE-1mRNA转录表达水平与胃溃疡的形成关系密切,冬胃颗粒对寒性胃溃疡具有较好的预防和治疗作用。
[Objective] To detect the effect of Dongwei Granule on the expression of ECE-1 in gastric mucosal cells of cold-evil type gastric ulcer in rats by real-time quantitative PCR and to explore the mechanism of Dongwei Granule in preventing and treating cold gastric ulcer at gene level. [Method] The rats model of gastric ulcer was prepared by injection of acetic acid and randomly divided into normal control group, ulcer model group, cold ulcer group, western medicine treatment group and traditional Chinese medicine treatment group. Cold ulcer model and treatment group were made ulcer model using cold factor to stimulate the replication of cold evil off stomach animal model, the first 3 days with 0 ℃ ice water soak, each 10min, 3 days after 0 ℃ ice water, The treatment group also given drug gavage. The gastric mucosal ulcer index (UI) and ulcer inhibition rate were determined. The content of ET-1 in gastric mucosa was detected by ELISA and the expression level of ECE-1 mRNA was detected by real-time quantitative PCR. [Results] The ulcer index of TCM treatment group was significantly lower than that of ulcer model group and cold ulcer group (P <0.01), and Western medicine treatment group (P <0.05), the ulcer inhibition rate was 54.22%, significantly higher than that of western medicine treatment group 45.40%. The level of ET-1 in ulcer model group and cold ulcer group was significantly higher than that in normal control group (P <0.01). The level of ET-1 in gastric mucosa of Chinese herbal medicine group was significantly lower than that of normal control group, and the difference was statistically significant (P <0.05, P <0.01) with the ulcer model group and cold ulcer group. Correlation analysis showed that the level of ET-1 in gastric mucosa was positively correlated with UI (r = 0.89, P <0.01). The expression level of ECE-1mRNA in gastric mucosa of ulcer model and cold ulcer model group was significantly increased (P <0.05, P <0.01), and the drug treatment could decrease the transcript level of ECE-1mRNA in gastric mucosa, (P <0.05), and the trend of ET-1 in gastric mucosa was the same (r = 0.91, P <0.01). [Conclusion] Cold stimulation can increase gastric mucosal lesion in rats with acetic acid-induced gastric ulcer. The level of ET-1 in gastric mucosa and the expression of ECE-1 mRNA in gastric ulcer rats are closely related to the formation of gastric ulcer. Dongwei granule has a good effect on cold gastric ulcer The prevention and treatment effect.