电针胃经经穴促进大鼠胃黏膜修复过程中相关蛋白磷酸化的研究

目的:探讨电针促胃黏膜修复过程中对相关蛋白磷酸化的影响。方法:20只SD大鼠分为空白组、模型组、针刺非穴组和针刺穴位组,每组5只。使用无水乙醇灌胃法制作大鼠胃黏膜损伤模型。针刺非穴组取"足三里"梁门"四白"穴旁的对照点,针刺穴位组取"足三里"梁门"四白"穴,每次电针30 min,每日1次,共治疗7 d。对大鼠的胃黏膜细胞分别进行磷酸化抗体蛋白芯片检测,同时筛选720种磷酸化蛋白的表达差异,并进行比较。结果:模型组胃黏膜损伤指数与空白组比较显著升高(P<0.01);针刺穴位组胃黏膜损伤指数低于模型组和针刺非穴组(P<0.01),而与空白组比较差异无统计学意义(P>0.05)。蛋白芯片分析示:针刺穴位组与针刺非穴组均可上调模型大鼠的蛋白磷酸化的水平(≥1.5倍),但针刺穴位组上调的蛋白大多数参与了细胞的增殖,而针刺非穴组只有几种蛋白参与细胞增殖;同时针刺穴位组与针刺非穴组还可下调蛋白磷酸化水平(≥1.5倍),且针刺组下调的蛋白参与抑制细胞凋亡,而针刺非穴组下调的蛋白基本不参与细胞活动。结论:电针足阳明胃经穴可引起大鼠胃黏膜损伤后修复信号蛋白的磷酸化水平发生变化,提示电针促进胃黏膜修复的机制与胃黏膜损伤后修复信号蛋白的磷酸化有关。

Observation on Changes of the Intragastric Protein Phosphorylation Level Involving Electroacupuncture-induced Improvement of the Injured Gastric Mucosa in the Rat

Objective To study the effect of electroacupuncture (EA) of Stomach Meridian acupoints on changes of the related protein phosphorylation in the gastric mucosa involving the repair of the injured gastric mucosa in the rat. Methods A total of 20 SD rats were divided into control, model, acupoint and non-acupoint groups, with 5 cases in each. Gastric mucosa injury model was established by intragastric perfusion of dehydrated alcohol (0. 8 ml/100 g). EA (4 Hz/50 Hz, 1-3 mA) was applied to "Zusanli" (ST 36), "Liangmen" (ST 21) and "Sibai" (ST 2), and the corresponding non-acupoints, once daily for 7 days. Gastric mucosal ulcer index (UI) was measured, and the profiling of the protein phosphorylation of interest in gastric mucosa cells was detected by antibody microarrays. Results In comparison with control group, the gastric mucosal Ul of model group was significantly higher than that of control group (P<0. 01), while the UI of EA group was significant lower than those of model group and non-acupoint group (P<0. 01), suggesting a recovery of the injured mucous membrane. Findings of protein array analysis indicated that a total of 720 kinds of phosphorylated proteins(≥1. 5 folds)were found in the injured gastric mucosa. Compared to model group, the phosphorylation levels of 100 and 20 proteins were up-regulated, and 16 and 9 proteins down-regulated in acupoint and non-acupoint groups, respectively. The up-regulated proteins involve cell cycle mediation, cell proliferation, anti-apop-tosis, and the regulation of the cell adhesion, cytoskeleton protein, inflammation, immune activities, etc. while the down-regulated proteins involve the down-regulation of the phosphorylation levels of the apoptosis proteins, adhesion proteins and cytoskeleton proteins, etc. In comparison to non-acupoint group, the phosphorylation levels of 100 proteins were up-regulated and 16 proteins down-regulated in acupoint group. Conclusion EA of acupoints of the Stomach Meridian can promote repair of the injured gastric mucosa, which may be related to its effects in regulating the levels of phosphorylation of many signaling proteins.