黄芪甲苷预处理对大鼠肠缺血再灌注损伤的影响

Effects of Astragaloside Ⅳ on Intestinal Ischemia-reperfusion Injury in Rats

目的:探讨黄芪甲苷(AS-IV)预处理对大鼠肠缺血再灌注损伤的影响。方法:30只成年雄性Wistar大鼠随机分为假手术(Sham)组、模型(I/R)组、AS-IV预处理(AS-IV)组。I/R组和AS-IV组采用无创微动脉夹夹闭肠系膜上动脉60min再灌注120min的方法建立大鼠肠缺血再灌注损伤模型。AS-IV组于造模前60min时采用60mg/kgAS-IV对大鼠进行灌胃预处理。于再灌注120min时取小肠组织,采用HE染色法测定小肠组织病理学变化,ELISA法测定大鼠血清中TNF-α、HMGB-1、SOD和CAT的表达情况,荧光定量PCR法测定小肠组织中Bcl-2、Bax和Caspase-9的mRNA情况,Western blotting法测定小肠组织中紧密连接蛋白occludin和ZO-1的表达情况。结果:与Sham组比较,I/R组小肠组织病理学Chiu评分升高,TNF-α和HMGB-1的表达水平升高,SOD和CAT的表达水平降低,Bcl-2、Bax和Caspase-9的mRNA水平升高,occludin和ZO-1的蛋白表达水平降低(P<0.05);与I/R组比较,AS-IV组I/R组小肠组织病理学Chiu评分降低,TNF-α和HMGB-1的表达水平降低,SOD和CAT的表达水平升高,Bcl-2、Bax和Caspase-9的mRNA水平降低,occludin和ZO-1的蛋白表达水平升高(P<0.05)。结论:AS-IV灌胃预处理能够显著改善大鼠肠I/R损伤,其机制可能与减轻机体炎症反应和氧化应激水平、抑制肠道细胞的凋亡发生以及调节紧密连接蛋白表达有关。

Objective To investigate the effects of astragaloside A(AS-IV)pretreatment on intestinal ischemia-reperfusion(I/R)injury in rats.Methods Thirty adult male Wistar rats were randomly divided into Sham group,model(I/R)group and AS-IV pretreatment(AS-IV)group.Intestinal I/R model was induced via clamping the superior mesenteric artery for 60min,then followed by 120 min reperfusion in I/R group and ASIV group.Rats received intragastric administration with 60 mg/kg AS-IV 60 min before ischemia in AS-IV group.After 120 min of reperfusion,pathological changes and score of the intestine were measured by HE staining;the levels of TNF-α,HMGB-1,SOD and CAT in serum were measured by ELISA assay;the mRNA levels of Bcl-2,Bax and Caspase-9 in intestinal tissues were measured by RT qPCR assay;the expression levels of occludin and ZO-1 were measured by Western blotting assay.Results Compared with Sham group,the Chiu score,levels of TNF-αand HMGB-1,and mRNA levels of Bcl-2,Bax and caspase-9 were increased,and levels of SOD and CAT and expression levels of occludin and ZO-1 were decreased in I/R group(P<0.05).Compared with I/R group,the Chiu score,levels of TNF-αand HMGB-1,and mRNA levels of Bcl-2,Bax and caspase-9 were decreased,and levels of SOD and CAT and expression levels of occludin and ZO-1 were increased in AS-IV group(P<0.05).Conclusion As-IV pretreatment can significantly alleviate intestinal I/R injury in rats,which is involved in lightening inflammation and oxidative stress,inhibiting apoptosis of intestinal cells,and regulating the expression of tight junction proteins.