灯盏花注射液通过抑制减IκB-α/NF-κB/ICAM-1通路轻大鼠肠缺血再灌注损伤

Breviscapine reduces intestinal ischemia reperfusion via inhibiting IκB-α/NF-κB/ICAM-1 pathway in a rat model

目的探讨灯盏花注射液在肠缺血再灌注早期小肠损伤中的作用及其机制。方法 48只8周龄健康雄性SD大鼠随机分为假手术对照(Sham)组,肠I/R损伤(IIR)组,灯盏花注射液处理组(EB+IIR)组,IκB-α抑制剂TPCA-1处理(TP+IIR)组。EB+IIR组在术前7 d每天经腹腔注射给予灯盏花注射液20 mg(/kg·d),TP+IIR组在术前30 min尾静脉给予TPCA-1(12 mg/kg),另两组注入等量生理盐水。采用夹闭SD大鼠肠系膜前动脉45 min后再灌注6h的方法造成肠缺血再灌注损伤模型。处死大鼠取小肠标本,HE染色观察肠组织病理学改变,ELISA检测肠粘膜组织TNF-α、IL-1β和IL-6水平;蛋白印迹法检测肠粘膜组织IκB-α、NF-κB、ICAM-1蛋白表达水平。结果肠缺血再灌注导致明显小肠损伤,肠组织炎症指标明显增高,表现为TNF-α、IL-1β和IL-6含量明显升高(与假手术组比较P<0.05),IκB-α/NF-κB/ICAM-1通路活化;灯盏花注射液预处理可抑制小肠组织IκB-α/NF-κB/ICAM-1活化,减轻小肠缺血再灌注引起的早期肠道损伤,并下调小肠组织TNF-α、IL-1β和IL-6含量(与缺血再灌注组比较P<0.05),同样,应用IκB-α磷酸化抑制剂TPCA-1处理后,IκB-α/NF-κB/ICAM-1通路受到抑制,小肠损伤程度得到减轻。结论灯盏花注射液预处理通过抑制IκB-α/NF-κB/ICAM-1通路减轻大鼠肠缺血再灌注损伤。

Objective To investigate the effects of breviscapine injection on intestinal injury induced by intestine ischemia-reperfusion （IIR）. Methods 48 males SD rats with 8-week old were randomly divided into 4 groups： Sham, intestine ischemia-reperfusion （IIR） , EB ＋ IIR, TP ＋ IIR. Breviscapine injection 20 mg/（kg, d） was given intraperitoneally in EB ＋ IIR group. TPCA-1 （12 mg/kg） was given intravenously 30 min before surgery in TP ＋ IIR group. Rats were subjected to superior mesenteric artery occlusion consisting of 45 rain of ischemia and 6 h of reperfusion ; sham laparotomy served as controls. Intestine pathology was assayed by H＆E staining. Con- centrations of TNF-α, IL-1β, and IL-6 in intestinal mucosa were determined by ELISA. The protein expressions of ⅠκB-α, NF-κB, ICAM-lof intestine tissue were assayed by western blot. Results IIR induced serious intesti- nal injury, evidenced as poor intestine pathology, elevation of TNF-α, IL-1β, and IL-6 levels in intestinal mu-cosa, accompanied with ⅠκB-α/NF-κB/ICAM-1 pathway activation. However, breviscapine injection pretreatment could inhibit ⅠκB-αNF-κB/ICAM-1 pathway activation, leading to reduction of TNF-α, IL-1β, and IL-6 concen- trations in lung, finally attenuate ALI induced by IIR. Conclusion Breviscapine injection pretreatment could atten- uate inflammation in intestine after IIR injury via inhibiting ⅠκB-α/NF-κB/ICAM-1signaling pathway.