中国实验动物学报
中國實驗動物學報
중국실험동물학보
ACTA LABORATORIUM ANIMALIS SCIENTIA SINICA
2014年
5期
17-21
,共5页
卢伟%李德芳%朱斌%吴靖平
盧偉%李德芳%硃斌%吳靖平
로위%리덕방%주빈%오정평
caspase-9抑制剂%胎牛血清%软骨终板%凋亡
caspase-9抑製劑%胎牛血清%軟骨終闆%凋亡
caspase-9억제제%태우혈청%연골종판%조망
Caspase-9 inhibitor%Fetal bovine serum%Cartilage endplate%Apoptosis Chondrocytes%Vertebra%Rat
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。
目的：探討caspase-9抑製劑對低胎牛血清培養誘導的大鼠椎間盤軟骨終闆細胞凋亡影響的研究。方法取3月齡SD大鼠椎間盤軟骨終闆，序貫消化法穫取細胞原代培養，以1％FBS培養48 h為誘導凋亡條件。實驗分為1％FBS凋亡組、caspase-9抑製劑組（ Z-LEHD-FMK）及DMSO對照組，分彆處理細胞48 h，後經流式細胞儀檢測細胞凋亡率、Western blot檢測procaspase-9，active caspase-9及active caspase-3的錶達。結果流式細胞儀檢測顯示，caspases-9抑製劑組細胞凋亡率（26.3±2.56）％與1％FBS組（40.8±0.84）％及DMSO組（40.2±1.56）％相比凋亡率較低，有顯著統計學差異（P＜0.05）；Western blot檢測caspases-9抑製劑組active caspase-9及active caspase-3較1％FBS凋亡組及DMSO對照組錶達均明顯減少，有顯著統計學意義（P＜0.05）。結論 Caspase-9抑製劑能明顯抑製低胎牛血清培養誘導的大鼠椎間盤軟骨終闆細胞凋亡，有望成為治療椎間盤退變的新型藥物。
목적：탐토caspase-9억제제대저태우혈청배양유도적대서추간반연골종판세포조망영향적연구。방법취3월령SD대서추간반연골종판，서관소화법획취세포원대배양，이1％FBS배양48 h위유도조망조건。실험분위1％FBS조망조、caspase-9억제제조（ Z-LEHD-FMK）급DMSO대조조，분별처리세포48 h，후경류식세포의검측세포조망솔、Western blot검측procaspase-9，active caspase-9급active caspase-3적표체。결과류식세포의검측현시，caspases-9억제제조세포조망솔（26.3±2.56）％여1％FBS조（40.8±0.84）％급DMSO조（40.2±1.56）％상비조망솔교저，유현저통계학차이（P＜0.05）；Western blot검측caspases-9억제제조active caspase-9급active caspase-3교1％FBS조망조급DMSO대조조표체균명현감소，유현저통계학의의（P＜0.05）。결론 Caspase-9억제제능명현억제저태우혈청배양유도적대서추간반연골종판세포조망，유망성위치료추간반퇴변적신형약물。
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 ±0.84)%and DMSO group (40.2 ±1.56)%, the apoptosis rate of the caspase-9 inhibitor group (26.3 ±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.
