HMGB1、TLR4在重症急性胰腺炎大鼠胰腺组织中的表达及乌司他丁的干预效应

目的：探讨 HMGB1、TLR4在重症急性胰腺炎大鼠胰腺组织中的作用机制以及乌司他丁的干预效应。方法将54只SD大鼠分为对照组、SAP组和乌司他丁治疗组，3组又分为6、12 h和24 h 3个小组（每小组n＝6）。对照组开腹后仅翻动胰腺组织，SAP组用5％的牛磺胆酸钠制备SAP模型，治疗组在SAP造模成功后经尾静脉注射乌司他丁。观察3组大鼠胰腺组织的病理学改变；EPS‐G7法检测血清中的淀粉酶；ELISA法检测血清及胰腺组织中的HMGB1；Envision两步免疫法检测胰腺组织中的HMGB1、TLR4的表达水平。结果 SAP组、治疗组各时间点的淀粉酶与对照组比较明显升高，病理学改变明显，差异均有统计学意义（P＜0．05），示SAP造模成功；SAP组在胰腺组织及血清中的 HMGB1表达在6 h开始升高，于12 h快速上升，至24 h保持上升趋势，与对照组大鼠相同时间点比较明显升高，差异有统计学意义（P＜0．05），治疗组与SAP组相同时间点的 HMGB1比较明显降低，差异有统计学意义（P＜0．05）；SAP组胰腺组织中的 TLR4表达在6 h开始升高，12 h达高峰，24 h开始下降，与对照组大鼠相同时间点比较明显升高，差异有统计学意义（P＜0．05）。治疗组与SAP组相同时间点的TLR4比较明显降低，差异有统计学意义（P＜0．05）。结论 HMGB1在SAP大鼠胰腺中的致炎作用可能是部分结合其受体 TLR4并通过MyD88依赖性途径而实现的，而乌司他丁可能是通过中断SAP大鼠胰腺组织中的 HMGB1、TLR4信号通路发挥保护作用。

The expression of HMGB1 and TLR4 in pancreatic tissue of rats with severe acute pancreatitis and the intervention effects of Ulinastatin

Objective To explore the mechanism of HMGB1 and TLR4 in pancreatic tissue of rats with severe acute pancreatitis and the intervention effect of Ulinastatin .Methods The 54 SD rats were completely random divided into control group ,SAP group and Ulinastatin treatment group ,and each group was divided into three groups :6 ,12 h and 24 h groups (each group n=6) .In con‐trol group ,we turned the pancreatic tissue ,in SAP group ,the SAP model was made with 5% taurocholic acid ;and in the treatment group ,and intravenous injection of ulinastatin was conducted after the SAP model was successfully made .Then we observed the pancreatic tissue pathology in the three groups .The amylase in serum was detected by EPS‐G7 assay ,the HMGB1 in serum and pancreatic tissue was detected by ELISA assay ,the expression levels of HMGB1 and TLR4 in pancreatic tissue were detected by Envision two‐step immunoassay .Results Compared with control group ,the amylase of each time point in SAP group and treatment group were significantly higher ,and the pathology changed obviously (P<0 .05) ,and the SAP model was successfully made .The HMGB1 expression in pancreatic tissue and serum started increase at 6 h ,increased quickly at 12 h and maintained the increasing trend to 24 h in SAP group and it was significantly higher at the same time point compared with that of control group (P<0 .05);at the same time point ,the HMGB1 in treatment group was significantly lower than that of SAP group (P<0 .05);in SAP group , the expression of TLR4 in pancreatic tissue started increasing at 6 h ,reached its peak at 12 h and started decreasing at 24 h ,it was significantly higher than the control group at the same time point (P<0 .05) .At the same time point ,the TLR4 was significantly lower in the treatment group than SAP group (P<0 .05) .Conclusion The proinflammatory effect of HMGB1 in SAP rats pancre‐atic could be partly combine its receptor TLR4 and MyD88‐dependent pathway through implementation ,and the protecting mecha‐nism of Ulinastatin could be interrupt the HMGB1 and TLR4 signaling pathway in SAP rats pancreatic tissue .