重症急性胰腺炎大鼠外周血中性粒细胞凋亡的变化及乌司他丁的干预作用

Effects of ulinastatin on polymorphonuclear neutrophil apoptosis in rats with severe acute pancreatitis

目的探讨重症急性胰腺炎(SAP)大鼠外周血中性粒细胞凋亡的变化及乌司他丁对不同凋亡通路的影响。方法健康Wistar大鼠90只随机分为3组:假手术组(SHAM组,30只)、SAP模型组(SAP组,30只)和乌司他丁治疗组(ULi组,30只)。大鼠经十二指肠乳头逆行注射3.5%牛黄胆酸钠诱导构建SAP模型;ULi组同法诱导构建SAP模型后,经门静脉给予乌司他丁50 000 U/kg;SHAM组开腹仅翻动胰腺和十二指肠。各组于术后0、3、6、12和24 h分批处死大鼠,切取胰腺组织,采集下腔静脉血。对各组不同时点大鼠胰腺组织进行胰腺组织病理学评分,测定血清淀粉酶,并分离中性粒细胞进行凋亡检测及Caspase1、3和8活性的检测。结果SAP组各时点胰腺组织病理学评分、血清淀粉酶水平及Caspase1活性明显高于SHAM组和ULi组(P<0.05),外周中性粒细胞凋亡率、Caspase3和8明显低于ULi组。结论中性粒细胞凋亡在SAP的病程及转归中发挥重要作用;乌司他丁对Caspase 3和8的活性有影响,可以显著减轻胰腺病理损伤,改善SAP预后。

Objective To explore the changes in polymorphonuclear neutrophils （PMN） and the effects of ulinastatin on different biomarker signaling pathways in rats with severe acute pancreatitis （SAP）. Methods Ninty healthy Wistar rats were randomly allocated to Sham group （n=30）, SAP group （n=30） and ulinastatin （ULi） group （n= 30）. The SAP rat model was induced by 3.15% sodium tauroeholate retrograde injection to the common biliopancreatic ducts via the papilla duodeni. Following establishment of the SAP rat model, rats in group ULi were treated with ulinastatin 50 000 U/kg through portal vein injection. Pancreas and duodenum were only flipped after opening the abdominal cavity in Sham group. Rats were sacrificed at 0, 3, 6, 12, 24 hours postoperatively for extraction of the pancreatic tissues and sampling of the blood from inferior vena cava. This entailed pathological scoring of the pancreas at different time points. Blood amylase was assayed. Apoptosis assay was conducted on the isolated PMN. And the Caspasel, 3 and 8 activity was quantified. Results Compared with Sham and ULi group, the levels of blood amylase, histopathologieal scores of the pancreas and caspase 1 activity were significantly increased, and the PMN apoptotic rate, caspase 3 and 8 activity were markedly lower in SAP group （all P〈0.05）. Conclusion PMN apoptosis rate may have played an important role in the pathogenesis and prognosis of SAP. Ulinastatin might improve the prognosis of SAP by attenuating the caspase 3 and 8 activity leading to mitigate pancreatic injury.