目的探讨大鼠急性胰腺炎时Notch-1的表达及其与胰腺细胞凋亡的关系。方法用不同浓度的牛磺胆酸钠逆行胆胰管注射分别建立急性轻型胰腺炎(MAP)及急性重型胰腺炎模型(SAP),采用TUNEL、Westernblot以及实时定量PCR的方法分别检测不同类型急性胰腺炎在不同时相的细胞凋亡指数、Notch-1蛋白以及Notch-lmRNA的表达水平变化。结果MAP组在建模后4~24h内细胞凋亡指数均明显升高,但各时点之间细胞凋亡指数相差不大;SAP组在建模4h后细胞凋亡指数最高,其后逐渐下降,两组之间细胞凋亡指数有显著性差异。建模后4h开始,各时点SAP组和MAP组的Notch1表达均有上升,但在SAP组的Notch-l表达明显高于MAP组,两组之间相比有显著差异;MAP组Notch-l表达的峰值时问是建模后12h,而SAP组Notch-1表达的峰值时间是建模后8h。结论不同类型急性胰腺炎细胞凋亡指数及Notch1的表达均有明显差异,Notch-1的过度表达可能通过抑制细胞凋亡而加重急性胰腺炎的病情。
Objective To investigate the relationship between cell apoptosis and Notch-1 expression in rats after acute pancreatitis. Methods Mild acute pancreatitis (MAP) and severe acute pancreatitis (SAP) models were established by retrograde injection of different dose sodium taurocholae into pancreatic duct, The apoptotic index, Notch-l mRNA and protein expression in two types of acute pancreatitis at different time phases were detected by TUNEL, Western blot and real-time PCR, respectively. Results In MAP group, the apoptotic index was elevated at 4 hour after induction of pancreatitis and retained at a relative high and stable level. In SAP group, the value of apoptotic index reached its peak at 4 hour after induction of pancreatitis and then declined gradually. The apoptotic index had significant difference between the 2 groups (P〈0.01). The expression of both Notch-l mRNA and protein was elevated after induction of pancreatitis in both groups at different time phases within 24 hour. However, the expression of Notch-l was significantly higher in SAP than in MAP(P 〈0.05). The peak of Notch 1 expression in MAP was at 12 hour after induction of pancreatitis and at 8 hour in SAP. Conclusion Both apoptotic index and Notch 1 expression are markedly different in different type of acute pancreatitis. Excess expression of Notch-l might aggravate acute pancreatitis through its anti-apoptotic effect,