经皮穴位电刺激对胃肠道手术致大鼠肠黏膜屏障损伤时炎症反应的影响

Effect of transcutaneous electrical acupoint stimulation on inflammatory responses during intestinal mucosal barrier damage caused by gastrointestinal surgery in rats

目的评价经皮穴位电刺激(TEAS)对胃肠道手术致大鼠肠黏膜屏障损伤时炎症反应的影响。方法清洁级健康雄性SD大鼠45只,7~8周龄,体重220~260 g,采用随机数字表法分为3组(n=15):对照组(C组)、胃肠手术组(S组)和TEAS组(T组)。大鼠行幽门切除+胃空肠Roux-en-Y吻合术制备胃肠道手术模型,C组不做处理。T组于术前30 min行双侧足三里和合谷穴经皮电刺激,以疏密波,电流强度5 mA,频率2 Hz刺激至术毕。S组仅贴敷电极贴片,不刺激。于术后12、24和48 h(T1-3)时每组随机处死5只大鼠,取回肠组织。光镜下观察病理学结果并行Chiu评分,采用ELISA法检测IL-6、TNF-α二胺氧化酶(DAO)和肠型脂肪酸结合蛋白(I-FABP)含量,Western blot法检测occludin、claudin-3和ZO-1表达,免疫组化法确定occludin、claudin和ZO-1阳性细胞百分比。结果与C组比较,S组和T组T1-3时Chiu评分、回肠IL-6、TNF-α、DAO和I-FABP含量升高,occludin、claudin-3和ZO-1阳性细胞百分比降低,occludin、claudin-3和ZO-1表达下调(P<0.05);与S组比较,T组T1-3时Chiu评分、回肠IL-6、TNF-α、DAO和I-FABP含量降低,occludin、claudin-3和ZO-1阳性细胞百分比升高,occludin、claudin-3和ZO-1表达上调(P<0.05)。结论TEAS可通过抑制炎症反应减轻胃肠道手术致大鼠肠黏膜屏障损伤。

Objective To evaluate the effect of transcutaneous electrical acupoint stimulation(TEAS)on inflammatory responses during intestinal mucosal barrier damage caused by gastrointestinal surgery in the rats.Methods Forty-five healthy male Sprague-Dawley rats,aged 7-8 weeks,weighing 220-260 g,were divided into 3 groups(n=15 each)by a random number table method:control group(C group),gastrointestinal surgery group(S group),and TEAS group(T group).Rats underwent pylorectomy and Roux-en-Y gastrojejunal anastomosis to establish gastrointestinal surgery models in S and T groups,while group C received no treatment.In group T,bilateral Zusanli and Hegu acupoints were stimulated with disperse-dense waves,current intensity 5 mA,frequency 2 Hz,from 30 min before surgery until the end of operation.Only electrode patches were applied,but no stimulation was applied in group S.Five rats were randomly selected from each group and sacrificed at 12,24 and 48 h after surgery(T1-3),and ileal tissues were taken after anesthesia.The pathological changes of ileal tissues were observed with a light microscope and scored according to Chiu.Enzyme-linked immunosorbent assay was used to determine contents of interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),diamine oxidase(DAO)and intestinal fatty acid-binding protein(I-FABP).Western blot was used to detect the expression of occludin,claudin-3 and ZO-1.The immunohistochemical method was used to determine the percentage of occludin,claudin and zonula occludens-1(ZO-1)positive cells.Results Compared with group C,Chiu′s scores and contents of ileal IL-6,TNF-α,DAO and I-FABP were significantly increased,the percentage of occludin,claudin-3 and ZO-1 protein positive cells was decreased,and the expression of occludin,claudin-3 and ZO-1 was down-regulated at T1-3 in group S and group T(P<0.05).Compared with group S,Chiu′s scores and contents of ileal IL-6,TNF-α,DAO and I-FABP were significantly decreased,the percentage of occludin,claudin-3 and ZO-1 protein positive cells was increased,and the expression of occludin,claudin-3 and ZO-1 was up-regulated at T1-3 in group T(P<0.05).Conclusion TEAS can reduce the intestinal mucosal barrier damage caused by gastrointestinal surgery by inhibiting the inflammatory response in rats.