川芎嗪通过抑制内质网应激减轻小肠缺血再灌注损伤的研究

目的研究川芎嗪(TMP)对大鼠小肠缺血再灌注损伤及内质网应激通路的影响。方法将120只Wistar大鼠按照随机数字表法平均分为假手术组、模型组和TMP低、中、高(15、30、60 mg/kg)剂量组。造模前30 min腹腔注射给药,假手术组、模型组腹腔注射生理盐水。通过夹闭肠系膜上动脉60 min制备小肠缺血再灌注大鼠模型,再灌注2 h后,检测肠组织含水量;通过HE染色法行肠组织病理学检查并进行损伤评分(Chiu′s氏评分)、TUNEL法观察细胞凋亡情况并计算凋亡指数(AI);ELISA法检测肠组织CRP、TNF-α、IL-1β含量;Western blot法检测内质网应激相关蛋白(GRP78、CHOP)及Cleved Caspase-12、Cleved Caspase-3、NF-κB蛋白表达。结果与模型组比较,TMP中、高剂量组大鼠肠组织含水量显著降低(P<0.05);肠系膜病变明显改善且凋亡细胞数量明显减少,Chiu′s氏评分和AI显著降低(P<0.01);肠组织CRP、TNF-α、IL-1β含量和GRP78、CHOP、Cleved Caspase-12、Cleved Caspase-3、NF-κB蛋白表达水平均显著降低(P<0.05)。结论TMP能够通过抑制内质网应激相关炎症和细胞凋亡通路,对小肠缺血再灌注损伤起到保护作用。

Tetramethylpyrazine attenuates intestinal ischemia-reperfusion injury by inhibiting endoplasmic reticulum stress

Objective To investigate the effects of Tetramethylpyrazine(TMP)on intestinal ischemia-reperfusion injury and endoplasmic reticulum stress pathway in rats.Methods 120 Wistar rats were randomly divided into sham operation group,model group,TMP low,medium,high dose(15,30,60mg/kg)group.The rat models with intestinal ischemia-reperfusion injury were made by clipping superior mesenteric artery for 60min.30min before surgery,the drugs were given by intraperitoneal injection,and the rats in sham operation group and model group were given normal saline.2h later,the water content of intestinal tissue was calculated;the mesentery pathological examination and cell apoptosis observation were performed by HE staining method and TUNEL method respectively,and the injury score(Chiu′s score)and apoptosis index(AI)were calculated;the content of CRP,TNF-α,IL-1βin intestinal tissue were detected by ELISA method;the expression of endoplasmic reticulum stress-related proteins(GRP78,CHOP),Cleved Caspase-12,Cleved Caspase-3,NF-κB were detected by Western blot method.Results Compared with model group,the water content in TMP medium,high dose group were significantly decreased(P<0.05);the mesenteric pathological changes was improved and the number of apoptotic cells was decreased significantly,the Chiu′s score and AI were significantly reduced(P<0.01);the expression of GRP78,CHOP,Cleved Caspase-12,Cleved Caspase-3,NF-κB were significantly down-regulated(P<0.05).Conclusion TMP can protect small intestine from ischemia-reperfusion injury by inhibiting endoplasmic reticulum stress-related inflammation and apoptosis.