Objective To investigate the effects of Necrostatin-1 ( Nec-1 ) in caerulein induced acute pancreatitis ( AP) in mice. Methods AP was induced by 7 peritoneal injections of caerulein (50 jxg/kg) at hourly intervals. Expression of RIP1 and RIP3 protein expressions were assessed by Western blot analysis at 3 h, 6 h, 9 h and 12 h after the first injection of caerulein. Another model of AP was induced by 10 peritoneal injections of caerulein at hourly intervals. Male C57BL/6 mice were divided randomly into 4 groups; Control group, Vehicle group, Nec-1 group and Nec-1 I group ( Nec-1 inhibitor). Nec-1 ( 1 mg/kg, q6h) , Nec-1 I and vehicle were also administered to mice by peritoneal injection at two hours before the first injection of caerulein. The severity of AP was determined by measuring,amylase and lipase activities in the serum and histological grading at 12 h, 18 h and 24 h after the first injection of caerulein. Levels of IL-1 (B and TNF-a mRNA in the pancreas were determined by real-time RT-PCR. Results Expression of RIP1 and RIP3 protein were markedly upregulated and correlated with acinar cell necrosis in the caerulein-induced murine model of AP. Compared with Vehicle and Nec-1 I groups, Nec-1 significantly reduced levels of serum amylase and lipase and pathological scores and levels of TNF-a and IL-1 p mRNAs in pancreatic tissues. Conclusion Nec-1 has remarkable protective effect against acute pancreatitis. Necroptosis may contribute to pancreatic injury in experimental pancreatitis.%目的 观察Necrostatin-1(Nec-1)对雨蛙肽诱导的小鼠急性胰腺炎(AP)的影响.方法 C57BL/6小鼠予以50 μg/kg的雨蛙肽腹腔注射7次,间隔1h,建立AP模型.于第1次雨蛙肽注射后3h、6h、9h和12h,取出胰腺组织,HE染色.Western blot检测RIP1和RIP3蛋白的表达;C57BL/6小鼠随机分为:Control组、Vehicle组、Nec-1组和Nec-li组,予以50 μg/kg的雨蛙肽腹腔注射10次,间隔1h,第1次雨蛙肽注射2h前,腹腔注射Nec-1(1mg/kg,每6h1次)、Nec-li或等体积溶剂和空白对照 于第1次雨蛙肽注射后12 h、18 h和24 h,收集血清和胰腺组织.检测血清淀粉酶和脂肪酶.HE染色评估胰腺损伤程度.Real-time RT-PCR检测胰腺组织IL-1 β和TNF-α mRNA的表达.结果 RIP1和RIP3蛋白在雨蛙肽诱导的AP中逐渐升高,且与胰腺腺泡细胞的坏死呈正相关;应用Nec-1后,与Vehicle组和Nec-li组比较,于12h、18 h和24 h,小鼠血清淀粉酶和脂肪酶水平明显降低,胰腺组织病理评分也明显减少,同时胰腺组织中IL-1β和TNF-α mRNA的水平也明显降低.结论 Nec-1对实验性胰腺炎具有明显的保护作用;程序性坏死可能促进了急性胰腺炎的损伤.
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