Caspase-9抑制剂对SD大鼠椎间盘软骨终板细胞凋亡的影响

目的探讨caspase-9抑制剂对低胎牛血清培养诱导的大鼠椎间盘软骨终板细胞凋亡影响的研究。方法取3月龄SD大鼠椎间盘软骨终板,序贯消化法获取细胞原代培养,以1%FBS培养48 h为诱导凋亡条件。实验分为1%FBS凋亡组、caspase-9抑制剂组（Z-LEHD-FMK）及DMSO对照组,分别处理细胞48 h,后经流式细胞仪检测细胞凋亡率、Western blot检测procaspase-9,active caspase-9及active caspase-3的表达。结果流式细胞仪检测显示,caspases-9抑制剂组细胞凋亡率（26.3±2.56）%与1%FBS组（40.8±0.84）%及DMSO组（40.2±1.56）%相比凋亡率较低,有显著统计学差异（P〈0.05）;Western blot检测caspases-9抑制剂组active caspase-9及active caspase-3较1%FBS凋亡组及DMSO对照组表达均明显减少,有显著统计学意义（P〈0.05）。结论 Caspase-9抑制剂能明显抑制低胎牛血清培养诱导的大鼠椎间盘软骨终板细胞凋亡,有望成为治疗椎间盘退变的新型药物。

Effect of caspase-9 inhibitor on apoptosis in cartilage endplate chondrocytes in rat vertebrae

Objective To explore the effect of caspase-9 inhibitor on low fetal bovine serum （ FBS）-induced apop-tosis in cartilage endplate chondrocytes in SD rat vertebrae.Methods Disc cartilage endplates were obtained from 3-month old SD rats and subjected to sequential digestion to harvest chondrocytes for primary culture, and apoptosis was in-duced by 1%FBS for 48 hours.Three groups of chondrocytes were treated by 1% FBS, caspase-9 inhibitor （ Z-LEHD-FMK） and DMSO, respectively.After 48 hours, apoptosis was detected by DAPI staining and flow cytometry.The expres-sion of procaspase-9, active caspase-9 and active caspase-3 was monitored by Western blot.Results Compared with the 1%FBS group （40.8 ＆#177;0.84）%and DMSO group （40.2 ＆#177;1.56）%, the apoptosis rate of the caspase-9 inhibitor group （26.3 ＆#177;2.56）% was significantly lower （P〈0.05）.The expressions of active caspase-9 and active caspase-3 in the caspase-9 inhibitor group were significantly lower than those in the other two groups （P〈0.05）.Conclusions Caspase-9 inhibitor can inhibit low FBS-induced apoptosis in cartilage endplate chondrocytes of rat vertebrae, and might become a new drug for the treatment of disc degeneration.