目的探讨细胞凋亡在急性胰腺炎炎症中的作用及受体相互作用蛋白(RIP)的调控作用。方法将30只C57小鼠分为3组:对照组、急性水肿性胰腺炎(AEP)组、急性坏死性胰腺炎(ANP)组。AEP组连续注射雨蛙素(50μg/kg)13次;ANP组连续注射雨蛙素(50μg/kg)13次,另注射1次脂多糖(15mg/kg);对照组注射等量生理盐水7次。末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记法观察胰腺腺泡细胞凋亡,实时荧光定量PCR法检测RIP1mRNA的表达,蛋白质免疫印迹法检测RIP1蛋白的表达。结果成功建立AEP及ANP小鼠模型。与对照组比较,2个胰腺炎模型组均存在细胞凋亡,且与AEP组比较,ANP组小鼠细胞凋亡减少,差异有统计学意义(P〈0.05)。与对照组比较,AEP组RIP1mRNA及蛋白表达均升高,而ANP组降低,差异均有统计学意义(P〈0.05)。结论 RIP1参与急性胰腺炎的发病过程,可能与调控腺泡细胞凋亡有关。
Objective To investigate the role of apoptosis and the regulating role of receptor interacting protein 1(RIP1)in acute pancreatitis.Methods Thirty C57 mice were divided into three groups:control group,acute edematous pancreatitis(AEP)group and acute necrotizing pancreatitis(ANP)group.The AEP group was continuously injected by cerulein 50μg/kg for 13 times,the ANP group was continuously injected by cerulein 50μg/kg for 13 times and lipopolysaccharide 15mg/kg once;the control group was injected by the same volume of normal saline for 7times.The acinar cell apoptosis was observed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling(TUNEL)assay.The RIP1 mRNA expression was measured by real time fluorescence PCR.The expression of RIP1 protein was detected by Western blotting.Results The mouse models of AEP and ANP were established successfully.Compared with the control group,acinar cell apoptosis existed in both AEP and ANP model groups,moreover compared with the AEP group,apoptosis in the ANP group were decreased,the differences were statistically significant(P〈0.05).Compared with the control group,the expression of RIP1 mRNA and protein in the AEP group was increased,while which in the ANP group were decreased,the differences were statistically significant(P〈0.05).ConclusionRIP1 participate in the pathogenesis of acute pancreatitis,which may associate with acinar cell apoptosis.