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电针对脓毒症大鼠肠黏膜机械屏障及紧密连接蛋白表达的影响。

Effect of electroacupuncture at ST36 on the intestinal mucosal mechanical barrier and expression of occludin in a rat model of sepsis.

机构信息

ICU Department, Zhejiang Chinese Medical University, Hangzhou, China.

出版信息

Acupunct Med. 2018 Oct;36(5):333-338. doi: 10.1136/acupmed-2016-011187. Epub 2018 Jul 12.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) at ST36 on the intestinal mucosal mechanical barrier and expression of the tight junction (TJ) protein, occludin, in a rat model of sepsis.

METHODS

60 male Wistar rats were randomly divided into six groups (n=10 rats each): Control, Control+EA, CLP (caecal ligation and puncture), CLP+EA, CLP+Sham-EA, and Sham-CLP. Rats of the CLP, CLP+EA and CLP+Sham-EA groups underwent CLP modeling of sepsis; those in the Sham-CLP underwent sham surgery and those in the Control and Control+EA groups remained unoperated. Rats in the CLP+EA and Control+EA groups received verum EA at ST36 and rats in the CLP+Sham-EA groups received EA at non-traditional acupuncture points. After three days, serum D-lactate concentrations were measured and ileal mucosa was collected for haematoxylin and eosin staining, morphological observation and Chiu's scoring. The intestinal epithelial cells were observed under transmission electron microscopy (TEM), while protein expression of occludin was measured by immunohistochemistry and Western blotting.

RESULTS

TJs of the Control, Sham-CLP and Control+EA groups were continuous under TEM but discontinuous in the CLP, CLP+EA and CLP+Sham-EA groups. Plasma D-lactate levels were significantly higher in the CLP, CLP+EA and CLP+Sham-EA groups compared with the Control, Sham-CLP and Control+EA groups (P<0.01). Protein expression of occludin, reflected by immunohistochemistry scores (IHS) and the results of Western blotting, were significantly reduced in the CLP, CLP+EA and CLP+Sham-EA groups when compared with the Control, Sham-CLP and Control+EA groups (P<0.01). Compared with the CLP group, the IHS and Western blotting results of the CLP+EA group were both significantly higher (P<0.05), while those of the CLP+Sham-EA group were similar to the CLP group.

CONCLUSIONS

Electrical stimulation at ST36 in rats with sepsis can increase protein expression of occludin, reduce serum D-lactate levels and increase permeability of the intestinal barrier.

摘要

目的

观察电针对脓毒症大鼠肠黏膜机械屏障及紧密连接(TJ)蛋白occludin表达的影响。

方法

60 只雄性 Wistar 大鼠随机分为 6 组(每组 10 只):对照组、对照组+电针(ST36)、盲肠结扎穿孔(CLP)组、CLP+电针组、CLP+假电针(Sham-EA)组和假手术(Sham-CLP)组。CLP 组、CLP+电针组和 CLP+Sham-EA 组大鼠行 CLP 法制作脓毒症模型;Sham-CLP 组大鼠行假手术,对照组和对照组+电针组大鼠不手术。CLP+电针组和对照组+电针组大鼠行 ST36 电针,CLP+Sham-EA 组大鼠行非传统穴位电针。3 天后,检测血清 D-乳酸浓度,取回肠黏膜行苏木精-伊红(HE)染色、形态学观察和 Chiu 评分。透射电镜(TEM)观察肠上皮细胞,免疫组化和 Western blot 检测 occludin 蛋白表达。

结果

TEM 下,对照组、Sham-CLP 组和对照组+电针组大鼠 TJ 连续,CLP 组、CLP+电针组和 CLP+Sham-EA 组大鼠 TJ 不连续。CLP 组、CLP+电针组和 CLP+Sham-EA 组大鼠血清 D-乳酸水平显著高于对照组、Sham-CLP 组和对照组+电针组(P<0.01)。CLP 组、CLP+电针组和 CLP+Sham-EA 组大鼠 occludin 蛋白表达显著低于对照组、Sham-CLP 组和对照组+电针组(P<0.01),免疫组化评分(IHS)和 Western blot 结果均提示 CLP+电针组大鼠 occludin 蛋白表达高于 CLP 组(P<0.05),而 CLP+Sham-EA 组与 CLP 组相似。

结论

电针对脓毒症大鼠 ST36 具有增强 occludin 蛋白表达、降低血清 D-乳酸水平和增加肠黏膜屏障通透性的作用。

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