中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
1期
39-41
,共3页
肖勇%孟庆良%舒晓刚%卢晓明%张维康
肖勇%孟慶良%舒曉剛%盧曉明%張維康
초용%맹경량%서효강%로효명%장유강
胰腺腺泡细胞%基因转染%脱噬作用
胰腺腺泡細胞%基因轉染%脫噬作用
이선선포세포%기인전염%탈서작용
Pancreatic acinar cells%Gene transfer%Apoptosis
目的 观察重组腺病毒介导的NICD基因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 (48 h)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Annexin V/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2.2×1010 pfu/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.
目的 觀察重組腺病毒介導的NICD基因對胰腺腺泡細胞凋亡的影響.方法 在293細胞中培養擴增重組腺病毒Ad-NICD,噹細胞彙閤度達到60％～70％時,用蝕斑形成實驗測定其感染滴度；將該病毒感染胰腺腺泡細胞AR4-2J (48 h)通過實時定量聚閤酶鏈反應(PCR)檢測其錶達.同時分對照組(未轉染組)、Ad-CMV轉染組、Ad-NICD轉染組分彆用Ad-CMV( MOI:100)和Ad-NICD( MOI:100)轉染AR4-2J細胞,採用Annexin V/PI雙標流式細胞術檢測Ad-NICD對細胞凋亡的影響.結果 重組腺病毒Ad-NICD滴度為2.2×1010 pfu/L,病毒顆粒感染腺泡細胞可以有效地提高NICD的錶達,在MOI為100時,NICD的錶達量最高.Ad-NICD感染胰腺腺泡細胞後,雨蛙素誘導的細胞凋亡率為(7.32±1.67)％,與對照組(18.55±2.61)％及Ad-CMV轉染組(19.76±4.28)％比較差異有統計學意義(P＜0.05).結論 重組腺病毒介導的NICD基因轉染可以顯著抑製雨蛙素誘導的胰腺腺泡細胞凋亡,這種抑製可能與急性胰腺炎的嚴重程度有關.
목적 관찰중조선병독개도적NICD기인대이선선포세포조망적영향.방법 재293세포중배양확증중조선병독Ad-NICD,당세포회합도체도60％～70％시,용식반형성실험측정기감염적도；장해병독감염이선선포세포AR4-2J (48 h)통과실시정량취합매련반응(PCR)검측기표체.동시분대조조(미전염조)、Ad-CMV전염조、Ad-NICD전염조분별용Ad-CMV( MOI:100)화Ad-NICD( MOI:100)전염AR4-2J세포,채용Annexin V/PI쌍표류식세포술검측Ad-NICD대세포조망적영향.결과 중조선병독Ad-NICD적도위2.2×1010 pfu/L,병독과립감염선포세포가이유효지제고NICD적표체,재MOI위100시,NICD적표체량최고.Ad-NICD감염이선선포세포후,우와소유도적세포조망솔위(7.32±1.67)％,여대조조(18.55±2.61)％급Ad-CMV전염조(19.76±4.28)％비교차이유통계학의의(P＜0.05).결론 중조선병독개도적NICD기인전염가이현저억제우와소유도적이선선포세포조망,저충억제가능여급성이선염적엄중정도유관.
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 adenoviruses were infected into acinar cells AR4-2J,and the expression of NICD was detected by using real-time polymerase 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 × 1010 pfu/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 NICD 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.
