CAJ | 학술논문

目的：观察不同浓度的苦杏仁苷对 IL-1β诱导的大鼠椎间盘软骨终板细胞凋亡的影响，并进一步探讨其作用的可能机制。方法从1月龄 SD 大鼠椎间盘中分离软骨终板并培养，经鉴定后，随机分为正常组、诱导组、苦杏仁苷10－2、10－3、10－4、10－5 mol·L －1给药组，采用流式细胞仪检测大鼠椎间盘软骨终板细胞的凋亡，Real-time PCR （RT-PCR）检测凋亡相关基因的表达情况，Western blot 检测细胞内 Bax、Bcl-2蛋白的表达。结果不同浓度的苦杏仁苷能够拮抗 IL-1β诱导的大鼠椎间盘软骨终板细胞凋亡。流式分析显示各浓度的苦杏仁苷可以降低 IL-1β诱导的大鼠椎间盘软骨终板细胞的凋亡比例；RT-PCR 结果提示苦杏仁苷各浓度组可拮抗 IL-1β上调 Bax mRNA 的表达，下调 Bcl-2 mRNA 的表达，各组与诱导组相比较，其差异具有统计学意义（P ＜0.05）；Western blot 检测结果显示苦杏仁苷10－4 mol·L －1给药组能够拮抗 IL-1β上调 Bax 蛋白的表达，下调 Bcl-2蛋白的表达。结论苦杏仁苷能够拮抗 IL-1β诱导大鼠椎间盘软骨终板细胞的凋亡，起到延缓椎间盘退变的作用。
목적：관찰불동농도적고행인감대 IL-1β유도적대서추간반연골종판세포조망적영향，병진일보탐토기작용적가능궤제。방법종1월령 SD 대서추간반중분리연골종판병배양，경감정후，수궤분위정상조、유도조、고행인감10－2、10－3、10－4、10－5 mol·L －1급약조，채용류식세포의검측대서추간반연골종판세포적조망，Real-time PCR （RT-PCR）검측조망상관기인적표체정황，Western blot 검측세포내 Bax、Bcl-2단백적표체。결과불동농도적고행인감능구길항 IL-1β유도적대서추간반연골종판세포조망。류식분석현시각농도적고행인감가이강저 IL-1β유도적대서추간반연골종판세포적조망비례；RT-PCR 결과제시고행인감각농도조가길항 IL-1β상조 Bax mRNA 적표체，하조 Bcl-2 mRNA 적표체，각조여유도조상비교，기차이구유통계학의의（P ＜0.05）；Western blot 검측결과현시고행인감10－4 mol·L －1급약조능구길항 IL-1β상조 Bax 단백적표체，하조 Bcl-2단백적표체。결론고행인감능구길항 IL-1β유도대서추간반연골종판세포적조망，기도연완추간반퇴변적작용。
Aim To observe the anti-apoptotic effect of different concentrations of amygdalin on the endplate chondrocytes induced by IL-1 βderived from rat inter-vertebral discs and explore the possible mechanism fur-ther.Methods Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary chondrocytes.After identifying,they were divided into normal group,induced group and A-mygdalin 1 0 -2 mol·L -1 ,1 0 -3 mol·L -1 ,1 0 -4 mol· L -1 ,1 0 -5 mol · L -1 administration group.Then the apoptosis was detected by flow cytometry (FCM).Re-al-Time PCR was adopted to detect the mRNA expres-sion of Bax and Bcl-2.The protein expression of Bax and Bcl-2 was detected by Western blot.Results The apoptosis of the endplate chondrocytes induced by IL-1 βderived from rat intervertebral discs could be inhib-ited by amygdalin with different concentrations.Amyg-dalin could reduce the apoptotic rate analysed by FCM,down-regulate the Bax mRNA expression of Bax and up-regulate the Bcl-2 mRNA assayed by RT-PCR;compared with the induced group the differences were statistically significant (P <0.05).Besides,observa-tion of the protein expression of Bax and Bcl-2 by Western blot found that amygdalin 1 0 -4 mol · L -1 could inhibit the effect of IL-1 β,which could increase the protein expression of Bax and reduce the protein expression of Bcl-2.Conclusion Amygdalin has an effect on anti-apoptosis of the end-plate chondrocytes induced by IL-1 βderived from rat intervertebral discs and improve the degeneration of intervertebral discs.