p38 MAPK抑制剂SB203580对雨蛙肽诱导的小鼠胰腺组织损伤的影响

摘要

AIM: To investigate the effect of p38 MAPK inhibitor SB203580 (SB) on caerulein (CAE)-induced injury in isolated mouse pancreatic tissues. METHODS: The pancreatic tissues of the mice were isolated, cultured for 4 h, and stimulated with CAE (10-5mol/L) alone or combined with SB (10-5mol/L). Normal saline (NS) was used as control reagent. At the time points of 1 h and 4 h after stimulation, the pancreatic tissues and the culture supernatants were harvested. The following parameters for evaluating the degree of injury were observed: the viability of the pancreatic tissues, the activity of amylase and lipase, and the content of interleukin-6 (IL-6) and cytokine-induced neutrophil chemo-tactic factor 1 (CINC-1) in the cultured supernatant. The levels of heat-shock protein (HSP) 60 and 70, and p38 and p-p38 proteins in the pancreatic tissues were measured by ELISA or Western blotting. RESULTS: After stimulation with CAE, the viability of the pancreatic tissues decreased at 1 h and was even lower at 4 h. The levels of amylase, lipase, IL-6, CINC-1, HSP60 and HSP70 as well as the expression of p38 and p-p38 were significantly increased at the time point of 1 h as compared with NS group. These changes were partly or totally prevented by SB treatment. CONCLUSION: CAE effectively damages the isolated pancreatic tissues. SB203580 reduces the injury, which may be related to the inhibition of the inflammatory responses.%目的:探讨p38 MAPK抑制剂SB203580(SB)对雨蛙肽(caerulein,CAE)诱导的小鼠离体胰腺组织损伤的影响,并探讨其可能的机制.方法:分离小鼠离体胰腺组织后培养4h,用CAE(10-5 mol/L)刺激,加用或不加用SB(10-5mol/L)进行干预,以生理盐水(NS)作为对照.在刺激1h和4h后测定胰腺组织活力(MTT)、培养上清液中淀粉酶和脂肪酶活性(生化法)以及白细胞介素6(IL-6)和细胞因子诱导的中性粒细胞趋化因子1(CINC-1)的水平(ELISA法)；同时测定胰腺组织中热休克蛋白60 (HSP60)和HSP70蛋白水平(ELISA法),并用Western blotting测定胰腺组织p38及磷酸化p38(p-p38) MAPK蛋白的表达.结果:CAE刺激后胰腺组织活力与NS组比较有所下降,尤其在刺激后4h明显降低(P ＜0.05)；CAE刺激后1h,离体胰腺组织培养上清液中淀粉酶、脂肪酶、IL-6和CINC-1水平较NS组均明显升高(P＜0.05)；胰腺组织中HSP60、HSP70、p38及p-p38 MAPK蛋白的表达较NS组有所升高(P＜0.05),而SB可不同程度干预CAE引起的这些指标的改变.结论:CAE对离体胰腺组织具有损伤作用,SB可减轻CAE造成的胰腺组织的损伤,其机制可能与其对p38 MAPK抑制进而使炎症反应受到抑制有关；HSP60和HSP70的变化是因为炎症反应减轻而使细胞应激程度降低所致,还是失去了p38 MAPK的调控作用所致,还有待进一步研究.