CAJ | 학술논문

目的：观察奥沙利铂（ OHP）对体外培养的背根神经节（ DRG）神经元的致凋亡作用，并探讨其机制。方法雌性Wistar大鼠雌性20只、雄性5只按4∶1合笼饲养，第2天选取孕鼠，至第12．5天（ E12．5）、19．5天（ E 19．5）时，将所获孕鼠麻醉后取出胚胎，体外培养DRG神经元。将选自E12．5及E19．5的胚胎DRG神经元各分为4组，在E12．5低、中、高剂量组及E12．5对照组培养液中分别加入10、20、50μmol/L OHP及正常细胞培养液；在E19．5低、中、高剂量组及E19．5对照组培养液中分别加入10、20、50μmol/L OHP及正常细胞培养液。各组均作用24 h。采用Hoechst33342染色DRG神经元细胞核，计算凋亡细胞所占比例。采用Western blot 法检测凋亡蛋白Caspase-3的表达。结果 E12．5中、高剂量组DRG细胞凋亡比例高于E12．5对照组及 E12．5低剂量组（ P＜0．05）。 E12．5中剂量组、E12．5高剂量组Caspase-3蛋白的表达高于E12．5对照组及E12．5低剂量组（P＜0．01），而低剂量组与对照组差异无统计学意义。结论 OHP可引起体外培养的E12．5 DRG神经元的凋亡，可能是通过干预DRG神经元的分化发挥致凋亡作用。
목적：관찰오사리박（ OHP）대체외배양적배근신경절（ DRG）신경원적치조망작용，병탐토기궤제。방법자성Wistar대서자성20지、웅성5지안4∶1합롱사양，제2천선취잉서，지제12．5천（ E12．5）、19．5천（ E 19．5）시，장소획잉서마취후취출배태，체외배양DRG신경원。장선자E12．5급E19．5적배태DRG신경원각분위4조，재E12．5저、중、고제량조급E12．5대조조배양액중분별가입10、20、50μmol/L OHP급정상세포배양액；재E19．5저、중、고제량조급E19．5대조조배양액중분별가입10、20、50μmol/L OHP급정상세포배양액。각조균작용24 h。채용Hoechst33342염색DRG신경원세포핵，계산조망세포소점비례。채용Western blot 법검측조망단백Caspase-3적표체。결과 E12．5중、고제량조DRG세포조망비례고우E12．5대조조급 E12．5저제량조（ P＜0．05）。 E12．5중제량조、E12．5고제량조Caspase-3단백적표체고우E12．5대조조급E12．5저제량조（P＜0．01），이저제량조여대조조차이무통계학의의。결론 OHP가인기체외배양적E12．5 DRG신경원적조망，가능시통과간예DRG신경원적분화발휘치조망작용。
Objective To investigate if oxaliplatin ( OHP) can induce apoptosis of dorsal root ganglion ( DRG) neu-rons and the mechanism of OHP-induced apoptosis .Methods In this study, we chose 20 female Wistar rats and 5 male rats, rearing for 4∶1 in five cages, and selected the female rats which had vaginal plug the next day .On embryonic 12.5 day (E12.5) and E19.5, the pregnant rats were anesthetized and then their embryos were removed to culture the DRG neurons in vitro.The cultured embryonic DRG neurons were divided into 4 groups respectively:E12.5 low, medium, high dose groups and E12.5 control group, which were incubated with 10, 20, 50 μmol/L OHP and normal cell culture fluid for 24 hours;E19.5 low, medium, high dose groups and E19.5 control group which were added 10, 20, 50μmol/L OHP and normal cell culture fluid for 24 hours.We used Hoechst33342 to stain the nucleus of DRG neurons and calculated the percentage of apoptosis cells .The expression of caspase-3 protein was detected by Western blotting .Results The percent-age of apoptosis cells in the E12.5 medium and high dose groups was higher than that of E 12.5 control and E12.5 low dose groups (P<0.05), and there was no significant difference between the low dose group and the control group .The expres-sion of caspase-3 protein in the E12.5 medium dose group and high dose group was higher than that of E 12.5 low dose group and E12.5 control group (P<0.01), but there was no significant difference between the low dose group and the control group.Conclusion OHP may induce the apoptosis of DRG neurons cultured in vitro by interfering the differentia-tion of DRG neurons .