姜黄素预处理对肠缺血-再灌注损伤的作用研究

Study on effects of curcumin pretreatment on intestinal ischemia-reperfusion injury

目的探讨姜黄素预处理对大鼠肠缺血-再灌注损伤(IRI)的作用及其机制。方法将30只SD大鼠随机均分为假手术组(Sham组)、IRI组和姜黄素预处理组(Cur组),每组10只。Cur组术前1 h给予姜黄素腹腔注射(5 mg/kg),Sham组和IRI组则给予等体积生理盐水腹腔注射,IRI组和Cur组行小肠IRI手术。观察3组大鼠肠组织病理学变化。采用逆转录聚合酶链反应法检测肠组织白细胞介素(IL)-6、IL-8和肿瘤坏死因子(TNF)-α的信使RNA(mRNA)水平。采用酶链免疫吸附试验(ELISA)检测血清中IL-6、IL-8和TNF-α的蛋白表达水平。采用硫代巴比妥酸反应物法检测肠组织丙二醛(MDA)含量,采用ELISA检测肠组织过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)含量。采用蛋白印迹法检测肠组织中磷脂酰肌醇-3-羟激酶(PI3K)、蛋白激酶(AKT)和哺乳动物雷帕霉素靶蛋白(m TOR)的蛋白表达水平。结果与Sham组相比,IRI组肠组织损伤程度,肠组织与血清中的IL-6、IL-8、TNF-α、MDA、PI3K、AKT、m TOR的表达水平均明显升高,而肠组织CAT、GPx与SOD含量明显降低。与IRI组相比,Cur组肠组织损伤程度,肠组织与血清中的IL-6、IL-8、TNF-α、MDA、PI3K、AKT、m TOR的表达水平均明显降低,而肠组织CAT、GPx与SOD含量明显升高(均为P<0.05)。结论姜黄素对大鼠发生IRI的肠组织有保护作用,该保护作用可能与抑制PI3K/AKT/m TOR信号通路介导的氧化应激和炎症有关。

Objective To explore the effects and mechanism of curcumin pretreatment on intestinal ischemiareperfusion injury（ IRI） in rats. Methods Thirty SD rats were randomly divided into sham operation group（ Sham group）,IRI group and curcumin pretreatment group（ Cur group）,with 10 rats in each group. Intraperitoneal injection with curcumin（ 5 mg / kg） was given to rats in Cur group at 1 h before operation; intraperitoneal injection with normal saline of same volume was given to rats in Sham group and IRI group. Rats in IRI group and Cur group received intestinal IRI operation. Pathological changes in intestinal tissues of the rats in 3 groups were observed. Messenger RNA（ mRNA） levels of interleukin（ IL）-6,IL-8 and tumor necrosis factor（ TNF）-α in intestinal tissue were detected by reverse transcription polymerase chain reaction,and protein expression levels of IL-6,IL-8 and TNF-α in serum were detected by enzyme linked immunosorbent assay（ ELISA）. Malondialdehyde（ MDA） in intestinal tissue was detected by thiobarbituric acid reactive substance assay,as well as contents of catalase（ CAT）,glutathione peroxidase（ GPx） and superoxide dismutase（ SOD）in intestinal tissue were detected by ELISA. Protein expression levels of phosphatidylinositol 3-kinase（ PI3K）,protein kinase（ AKT） and mammalian target of rapamycin（ m TOR） in intestinal tissue were detected by Western blotting. Results Compared with Sham group,the injury degree of intestinal tissue as well as the expression levels of IL-6,IL-8,TNF-α,MDA,PI3 K,AKT and m TOR in intestinal tissue and serum increased significantly,while the contents of CAT,GPx andSOD in intestinal tissue decreased significantly in IRI group. Compared with IRI group,the injury degree of intestinal tissue as well as the expression levels of IL-6,IL-8,TNF-α,MDA,PI3 K,AKT and m TOR in intestinal tissue and serum decreased significantly,while the contents of CAT,GPx and SOD in intestinal tissue increased significantly in Cur group（ all in P 〈0. 05）. Conclusions Curcumin presents protective effect on intestinal tissue with IRI in rats,which may be related to the inhibition of oxidative stress and inflammation mediated by PI3 K / AKT / m TOR signaling pathway.