电针促进胃黏膜损伤修复的时效关系及分子机制

Time-effect relationship and molecular mechanism of electroacupuncture for repair of gastric mucosal lesion

目的:动态观察电针对胃溃疡模型大鼠胃黏膜损伤修复的影响,探讨电针治疗胃溃疡的时效关系和分子机制。方法:72只SD大鼠分为空白组、模型组、胃经穴组、对照点组,并按干预时间1、4、7 d分为3个亚组,每个亚组6只。采用乙醇灌胃方法制备胃溃疡大鼠模型,胃经穴组电针"足三里""梁门",对照点组电针"梁门""足三里"外旁开5 mm处,每日1次,每次电针30 min。空白组和模型组用鼠板束缚但不进行电针处理,每日1次,每次30 min。用逆转录-聚合酶链反应(PR-PCR)法检测胃增殖细胞核抗原(PCNA)、P物质(SP)的表达,Western blot检测胃神经降压素(NT)的表达。结果:干预1 d后,模型组溃疡指数显著高于空白组(P<0.01),PCNA、SP、NT表达降低(P<0.01,P<0.05);胃经穴组与模型组、对照点组相比,溃疡指数降低(均P<0.05),上调PCNA m RNA、SP m RNA的表达(均P<0.05)且显著上调NT蛋白表达(均P<0.01)。干预4 d后,模型组溃疡指数降低但仍高于空白组(P<0.05),PCNA m RNA、SP m RNA、NT蛋白的表达上升且高于空白组(均P<0.01);胃经穴组溃疡指数接近空白组(P>0.05),PCNA m RNA、SP m RNA低于模型组(P<0.01,P<0.05),NT蛋白与模型组比较差异无统计学意义(P>0.05)。干预7 d后,各组以上指标组间比较差异均无统计学意义(均P>0.05)。结论:电针胃经穴能在胃溃疡发展的不同病理状态下对PCNA m RNA、SP m RNA进行双向调节的平衡作用同时促进NT蛋白高表达,进而促进溃疡的修复。

Objective To dynamically observe the effects of electroacupuncture（EA） on repair of gastric mucosal lesion in rats with gastric ulcer, and to explore the time-effect relationship and molecular mechanism of EA for gastric ulcer. Methods A total of 72 SD rats were randomly assigned to a normal group, a model group, a acupoint group and a sham acupoint group, and each group were further divided into a 1-day subgroup, a 4-day subgroup and a 7-day subgroup, 6 rats in each subgroup. The rat model of gastric ulcer was established by using intragastric administration of ethyl alcohol. The rats in the acupoint group were treated with EA at＂Zusanli＂（ST 36） and＂Liangmen＂（ST 21）; the rats in the sham acupoint group were treated with EA at points 5 mm next to＂Zusanli＂（ST 36） and＂Liangmen＂（ST 21）; the EA was given 30 min per treatment, once a day. The rats in the normal group and model group were treated with immobilization for 30 min per day, and no EA was given. PR-PCR method was applied to test the expression of proliferating cell nuclear antigen（PCNA） and substance P（SP）; Western blot method was applied to test the expression of neurotensin（NT）. Results After 1-day treatment, the ulcer index in the model group was higher than that in the normal group（P〈0.01）, and the expression of PCNA, SP and NT was decreased（P〈0.01, P〈0.05）; compared with the model group and sham acupoint group, the ulcer index was decreased in the acupoint group（both P〈0.05）, and the expression of PCNA and SP was up-regulated（all P〈0.05） while that of NT was up-regulated（both P〈0.01）. After 4-day treatment, the ulcer index in the model group was reduced but still higher than that in the normal group（P〈0.05）, and the expression of PCNA, SP and NT was up-regulated and higher than that in the normal group（all P〈0.01）; the ulcer index in the acupoint group was similar to that in the normal group（P〉0.05）, and the expression of PCNA and SP was lower than that in the model group（P〈0.01, P〈0.05）, and the expression of NT was not significantly different from that in the model group（P〉0.05）. After 7-day treatment, the differences of indexes above were not significant among the four groups （all P〉0.05）. Conclusion EA at acupoints of stomach meridian has two-way regulation on PCNA and SP and improve the expression of NT in different pathological state of gastric ulcer, which could further improve the repair of gastric ulcer.