NICD基因转染对胰腺腺泡细胞凋亡的影响  

Anti-apoptosis effects of NICD gene transfer to pancreatic acinar cells

作　　者：肖勇[1] 孟庆良[1] 舒晓刚[1] 卢晓明[1] 张维康[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院胃肠外科,武汉430022

出　　处：《中华实验外科杂志》2012年第1期39-41,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察重组腺病毒介导的NICD基冈对胰腺腺泡细胞凋亡的影响。方法在293细胞中培养扩增重组腺病毒Ad—NICD，当细胞汇合度达到60％～70％时，用蚀斑形成实验测定其感染滴度；将该病毒感染胰腺腺泡细胞AR4—2J（48h）通过实时定量聚合酶链反应（PCR）检测其表达。同时分对照组（未转染组）、Ad—CMV转染组、Ad—NICD转染组分别用Ad—CMV（MOI：100）和Ad—NICD（MOI：100）转染AR4—2J细胞，采用AnnexinV／PI双标流式细胞术检测Ad．NICD对细胞凋亡的影响。结果重组腺病毒Ad—NICD滴度为2．2×10^10pfu／L，病毒颗粒感染腺泡细胞可以有效地提高NICD的表达，在MOI为100时，NICD的表达量最高。Ad—NICD感染胰腺腺泡细胞后，雨蛙素诱导的细胞凋亡率为（7．32±1．67）％，与对照组（18．55±2．61）％及Ad—CMV转染组（19．76±4．28）％比较差异有统计学意义（P〈0．05）。结论重组腺病毒介导的NICD基因转染可以显著抑制雨蛙素诱导的胰腺腺泡细胞凋亡，这种抑制可能与急性胰腺炎的严重程度有关。Objective To investigate effect of the recombinant adenovirus Ad-NICD on apoptosis of pancreatic acinar cells in vitro. Methods The recombinant adenovirus Ad-NICD was propagated in 293 cells and the titer of adenovirus was determined by plaque bacteriophage formation test. The adenovimses were infected into acinar cells AR4-2J, and the expression of NICD was detected by using real-time poly-merase chain reaction （PCR）. Control group, Ad-CMV-transfected group and Ad-NICD-transfected group were set up. Apoptosis of the infected acinar cells was induced by cerulein and the effect of Ad-NICD was detected by Annexin V/PI double labelling cytometry. Results The titer of adenovirus was 2. 2×10^l0pfu/L and the expression of NICD was increased effectively after the acinar cells were infected by the virus. When MOI was 100, the expression of N1CD was highest. After treatment with cerulein, the apoptosis index of acinar cells in Ad-NICD-transferred group was （7. 32±1.67 ） %, which was significantly lower than in control group （ 18.55±2. 61 ） % and Ad-CMV-transferred group （ 19. 76±4. 28） %. Conclusion Recombinant adenovirus-mediated gene transfer of NICD could inhibit the apoptosis of pancreatic acinar cells induced by cerulien, which may be related to the severity of acute pancreatitis.

关 键 词：胰腺腺泡细胞 基因转染 脱噬作用 

分 类 号：R5[医药卫生—内科学]