p38 MAPK抑制剂SB203580对雨蛙肽诱导的小鼠胰腺组织损伤的影响

Effect of p38 MAPK inhibitor SB203580 on caerulein-induced pancreatic tissue injury in mice

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4 h,用CAE(10-5mol/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1 h和4 h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Westernblotting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4 h明显降低(P<0.05);CAE刺激后1 h,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。

AIM： To investigate the effect of p38 MAPK inhibitor SB203580 （SB） on eaerulein （CAE）-in- duced injury in isolated mouse pancreatic tissues. METItODS ： The pancreatic tissues of the mice were isolated, cultured for 4 h, and stimulated with CAE （10-~mol/L） alone or combined with SB （10-Stool/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 HSP＇/0 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.