罗格列酮对重症急性胰腺炎大鼠胰腺氧化应激的影响

 目的 探讨罗格列酮(rosiglitazone,ROSI)对重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠胰腺组织氧化应激的影响。方法 雄性SPF级SD大鼠60只,按随机数字表法随机分为3组:假手术组(sham operation group,SO组)、SAP组、ROSI组,每组20只。胆胰管逆行注射5%牛磺胆酸钠制备SAP模型。ROSI组在造模后5 min经股静脉注射ROSI(6 mg/kg),SO组和SAP组则注射等量生理盐水。术后12 h剖杀大鼠,统计各组大鼠死亡率。检测血清淀粉酶(amylase,AMY);检测胰腺组织一氧化氮(nitric oxide,NO)、丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平;胰腺组织行病理学检查;酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测血清白介素-1β(interleukine-1β,IL-1β)、白介素-6(interleukine-6,IL-6)水平。免疫印迹法(western blotting)检测胰腺组织诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、内皮细胞型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)蛋白水平。结果 SAP组死亡2只大鼠;SO组和ROSI组无大鼠死亡。SAP组大鼠AMY、IL-1β、IL-6、NO、MDA、胰腺病理评分均较SO组明显升高,差异有统计学意义(P<0.05);ROSI组干预后上述指标较SAP组明显下降,差异有统计学意义(P<0.05)。SAP组GSH-Px水平较SO组明显下降,给予ROSI干预后GSH-Px水平有所上升,但仍低于SO组[(49.01±9.42) U/mg vs (59.81±22.43 )U/mg],差异均有统计学意义(P<0.05)。SO组胰腺组织iNOS和eNOS蛋白呈低表达,SAP组iNOS和eNOS蛋白表达水平较SO组明显升高[(1.90±0.16 )vs(0.75±0.09)、(0.37±0.09)vs(0.09±0.01)],差异有统计学意义(P<0.05),ROSI干预后其表达水平均下降[(1.31±0.18)vs(1.90±0.16)、(0.23±0.08)vs(0.37±0.09)],差异有统计学意义(P<0.05)。结论 罗格列酮对SAP大鼠胰腺损伤具有保护作用,其机制与抑制氧化应激、减少炎性反应因子产生有关。

Effect of rosiglitazone on oxidative stress in rats with severe acute pancreatitis

Objective To investigate the effect of rosiglitazone on oxidative stress in rats with severe acute pancreatitis.Methods Sixty male SD rats were randomly divided into 3 group(n=20): sham operation group (SO group), severe acute pancreatitis group(SAP group) and rosiglitazone treatment group (ROSI group). SAP model was induced by retrograde infusion of 5% sodium taurocholate into the biliopancreatic duct. The rats in ROSI group were injected rosiglitazone(6 mg/kg) throngh femoral vein 5 minutes after the operation. In SO group and SAP group saline partes equales were injected. Rats were killed at 12 h after operation. Analysis of the mortality of rats in each group was conducted. Detecting amylase(AMY), nitric oxide(NO), malondialdehyde(MDA)、glutathione peroxidase(GSH-Px), pancreas pathologic score. Enzyme-linked immunosorbent assay (ELISA)and analysis of serum interleukine-1β (IL-1β), interleukine-6 (IL-6) levels were carried out. The expression of inducible nitric oxide synthase(iNOS)、endothelial nitric oxide synthase(eNOS)in pancreas tissue were measured by Western blotting method.Results Two rats died in SAP group, no rat died in SO group and ROSI group. Compared with SO group, the levels of AMY, NO, MDA and pancreas pathologic score were significantly increased in SAP group.The level of GSH-Px activity decreased significantly in SAP group than in SO group[(39.93±15.18) vs (59.81±22.43) U/mg], which was higher in ROSI group than in SAP group[(49.01±9.42) vs (59.81±22.43) U/mg] (P<0.05). The expression of iNOS and eNOS in pancreas tissue increased significantly in SAP group than in SO group[(1.90±0.16) vs (0.75±0.09),(0.37±0.09) vs (0.09±0.01)] (P<0.05); compared with SAP group, these indexes decreased significantly in ROSI group[(1.31±0.18) vs (1.90±0.16)、(0.23±0.08) vs (0.37±0.09)] (P<0.05).Conclusions Rosiglitazone exerts the protective effect on severe acute pancreatitis through inhibiting oxidative stress and downregulating the expression of inflammatory cytokine.