PI3K抑制剂对急性胰腺炎细胞因子和组织病理学评分的影响

Effect of PI3K inhibitor wortmannin on cytokine levels and pancreatic histopathological scores in rats with severe acute pancreatitis

目的:研究磷脂酰肌醇-3激酶(phosphatidylino-sitol 3-kinase,P13K)/丝/苏氨酸激酶(ser-ine threonine kinase,Akt)信号通路抑制剂渥曼青霉素(wortmannin)对重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠细胞因子[肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、IL-6]转录和表达以及胰腺组织病理评分的影响,探讨其对SAP大鼠的保护作用.方法:将60只SD大鼠随机分为重症急性胰腺炎组(S组)、假手术组(C组)、生理盐水组(NS组)、溶剂(DMSO)对照组(R组)、PI3K/Akt抑制剂wortmannin组(W组).采用改良的Aho法制作SAP模型,分别在造模3、6h后抽血并处死大鼠,收集胰腺组织.采用酶联免疫吸附试验(ELISA)检测血清细胞因子(TNF-α、IL-1β、IL-6)表达.Real-time PCR方法检测胰腺组织细胞因子(TNF-α、IL-1β、IL-6)mRNA转录水平变化,并常规HE染色进行胰腺组织病理评分,同时观察各组腹水量、血清及腹水淀粉酶的变化.结果:S组和R组大鼠术后3、6h的腹水量、血清及腹水淀粉酶水平、胰腺组织病理评分较C组和NS组差异有统计学意义(P<0.05);W组的腹水量、血清及腹水淀粉酶水平、胰腺组织病理评分较同时段的S组和R组有所降低,差异有统计学意义(P<0.05).与C组和NS组相比,S组和R组大鼠术后3、6h的血清细胞因子TNF-α、IL-1β、IL-6表达水平和胰腺组织内TNF-α、IL-1β、IL-6mRNA转录水平差异有统计学意义(P<0.05);W组的上述指标均低于S组和R组,差异有统计学意义(P<0.05).结论:PI3K抑制剂wortmanin通过抑制PI3K/Akt信号转导通路下调SAP大鼠细胞因子(TNF-α、IL-1β、IL-6)的转录和表达水平,减轻胰腺组织的病理损伤,对SAP大鼠具有保护作用.

AIM：To investigate the effect of the PI3K/Akt signaling transduction pathway inhibitor wortmannin on the levels of cytokines [tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β） and IL-6] and pancreatic histopathological changes in rats with severe acute pancreatitis（SAP）.METHODS：Sixty male Sprague-Dawley（SD） rats were randomly divided into five groups,including SAP group,sham operation group,normal saline group,DMSO control group,and wortmannin group.The modified Aho＇s method was used to reproduce the SAP model.The rats were sacrificed 3 and 6 h after treatment.The levels of inflammatory cytokines TNF-α,IL-1β and IL-6 in serum were determined by ELISA.Transcription levels of these inflammatory cytokines in pancreatic tissue were determined by real-time PCR.In addition,the amount of ascites,the activities of serum amylase and ascites amylase,and the pathological scores of pancreatic tissue were also measured.RESULTS：At 3 and 6 h after treatment,all parameters tested,including the amount of ascites,the levels of serum and ascites amylase,the pathological scores of pancreatic tissue,serum levels of TNF-α,IL-1β and IL-6,and the transcription levels of TNF-α,IL-1β and IL-6 mRNAs in the pancreatic tissue,in the SAP group and DMSO group were significantly higher than those in the normal saline group and sham operation group（all P〈0.05）.Compared to the SAP group and DMSO group,the above parameters decreased significantly in the wortmannin group（all P〈0.05）.CONCLUSION：Wortmannin exerts a protective effect against SAP possibly by down-regulating the levels of TNF-α,IL-1β and IL-6 and reducing pathological injury of pancreatic tissue in rats.