热休克蛋白27在大鼠急性胰腺炎中的表达及其对细胞间连接的影响

Effects of HSP27 expression on the adhesion junction of cells during acute pancreatitis in rats

目的探讨热休克蛋白27(HSP27)在轻型急性胰腺炎(MAP)及重症急性胰腺炎(SAP)大鼠模型中的表达及其对腺泡细胞间连接的影响。方法雄性Wistar大鼠90只,随机分3组:A组为SAP组,B组为MAP组,C组为对照组。3组动物于成模后1,3,8h分批处死。检测血清淀粉酶(SAM),计算胰腺系数;Western-blot半定量分析HSP27蛋白和磷酸化蛋白含量;RT-PCR半定量分析HSP27 mRNA表达;透射电镜观察细胞间连接的变化。结果A,B组SAM水平升高,但B组明显低于A组(P<0.05);A组胰腺系数迅速升高并持续增长,间质水肿严重(P<0.05),细胞间连接损伤;B组间质水肿比A组轻(P<0.05),细胞连接比较稳定;A,B两组HSP27蛋白及其mRNA表达均显著高于对照组(P<0.05),B组明显高于A组(P<0.05);B组HSP27磷酸化蛋白(Ser78)含量显著高于对照组和A组(P<0.05),A组与对照组比较无明显差异(P>0.05)。结论细胞间连接破坏是急性胰腺炎的共同特征,但SAP损伤更严重,可能与细胞间连接调控因子HSP27及其磷酸化表达相对下降。提示HSP在AP发生、发展中具有重要作用。

Objective To explore the effects of heat shock protein 27 （HSP27） expression on adhesion junction of cells during severe acute pancreatitis （SAP） and mild acute pancreatitis （MAP） in rats. Methods Ninty male Wistar rats were divided randomly into three groups： Group A （SAP）, Group B （ MAP ） and group C （ control ） . Rats in the three groups were kiUed at 1 h, 3 h. and 8 h after model made respectively. The levels of serum amylase （SAM） and histopathology score were measured. Changes of adhesion junctions were examined by electron microscopy. HSP27 and its phosphorylation were measured by Westeru-blotting. The expression of HSP27 mRNA was determined by RT-PCR. Results Levels of SAM increased in group A and group B, but was markedly lower in Group B （ P 〈 0.05 ） . In group A, there was a rapid and sustained increase in levels of pancreatic indexes, and severe interstitial edema （ P 〈 0.05 ） , and damages of adhesion junctions were observed. In group B, there was moderate pancreatic edema and disruption of adhesion junctions. The expression of HSP27, its mRNA and its phosphorylation levels were lower in group A than those in group B （P 〈 0.05）. Conclusions Alterations of adhesion junction are a common feature of AP, but the damage of HSP27 and its phosphorylation which regulate adhesion junctions were more severe during SAP. The results suggest that HSP plays an important role in the onset and development of AP