摘要
目的研究沉默DUSP1基因对急性胰腺炎（AP）小鼠促炎因子释放的调控机制。方法设计2条DUSP1-siRNA序列和1条阴性对照序列，选择沉默效率高的DUSP1-siRNA2序列。建立AP小鼠模型，分为6组：对照组、AP组、AP＋PD98059组、AP＋干扰组、AP＋siRNA组和AP＋PD98059＋siRNA组。建模成功后用酶联免疫吸附试验（ELISA）检测血清中促炎因子肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β和IL-6的表达，同时检测血清淀粉酶水平。采用定量实时聚合酶链反应（qPCR）法检测胰腺组织中TNF-α、IL-1β和IL-6水平，免疫印迹法检测胰腺组织中DUSP1、细胞外调节蛋白激酶（ERK）、c-Jun氨基末端激酶（JNK）、p38、p-ERK、p-JNK和p-p38的表达水平。结果与对照组相比，其他5组血清TNF-α、IL-1β、IL-6和血清淀粉酶及组织中DUSP1、TNF-α、IL-1β、IL-6、p-ERK、p-JNK和p-p38的表达均升高，差异均有统计学意义（均P〈0.05）。与AP组相比，AP＋PD98059＋siRNA组胰腺组织中DUSP1的表达下降，差异均有统计学意义（P〈0.05），AP＋PD98059组血清TNF-α、IL-1β、IL-6和淀粉酶的表达下降，胰腺组织中TNF-α、IL-1β、IL-6、p-ERK、p-JNK、p-p38的表达也下降，差异均有统计学意义（均P〈0.05）；而在DUSP1表达下降的AP＋siRNA组中结果却相反。结论沉默DUSP1基因可以激活促分裂原活化蛋白激酶（MAPK）信号通路，从而促进AP小鼠模型中促炎因子的释放。
ObjectiveTo investigate the regulation mechanism of silencing the DUSP1 gene on the release of proinflammatory cytokines in mice with acute pancreatitis（AP）.MethodsTwo DUSP1-siRNA and one scramble siRNA sequences were designed, and the sequence with higher silence efficiency was selected.Mice models with AP were established, and KM mice were divided into 6 groups： control group, AP group, AP＋ PD98059 group, AP＋ scramble group, AP＋ siRNA group and AP＋ PD98059＋ siRNA group.Expressions of proinflammatory cytokines tumor necrosis factor-α（TNF-α）, interleukin（IL）-1β and IL-6 in serum were detected by using enzyme linked immunosorbent assay（ELISA） after 12 h, 24 h, 48 h of modeling.Serum amylase levels were detected.The mRNA expression levels of DUSP1, TNF-α, IL-1β and IL-6 in pancreatic tissues were detected by using quantitative real time polymerase chain reaction （qPCR）. The protein expression levels of DUSP1, extracellular regulated protein kinases（ERK）, c-Jun N-terminal kinase（JNK）, p38, p-ERK, p-JNK and p-p38 in pancreatic tissues were detected by using Western blot.ResultsCompared with the control group, the other 5 groups displayed the increased expressions of TNF-α, IL-1β, IL-6 and amylase in serum, and expressions of DUSP1, TNF-α, IL-1β, IL-6, p-ERK, p-JNK, p-p38 in tissues, and there was a statistical significance （all P〈0.05）. Compared with the AP group, the AP＋ PD98059＋ siRNA group showed the decreased DUSP1 expression in tissues, and there was a statistical significance （all P〈0.05）; the AP＋ PD98059 group had decreased expressions of TNF-α, IL-1β, IL-6 and amylase in serum, and expressions of TNF-α, IL-1β, IL-6, p-ERK, p-JNK, p-p38 in tissues, and there were statistical significances （all P〈0.05）; while the opposite results were observed in the AP＋ siRNA group with DUSP1 expression decreased.ConclusionsThe results support that silencing the DUSP1 gene promotes the release of proinflammatory cytokines through activating the mitogen-activated protein kinase signaling pathway in mice with AP.
作者
齐凤芹
张波
Qi Fengqin;Zhang Bo(Department of Pediatrics,Liaocheng Second People′s Hospital,Linqing 252600,Shandong Province,Chin;Department of Emergency,Liaocheng Second People′s Hospital,Linqing 252600,Shandong Province,China)
出处
《中华实用儿科临床杂志》
CSCD
北大核心
2018年第19期1478-1482,共5页
Chinese Journal of Applied Clinical Pediatrics
