苦杏仁苷联合羟基红花黄色素A对IL-1β诱导的大鼠椎间盘软骨终板细胞的影响  

The synergistic effect of amygdalin and HSYA on the IL-1β induced endplate chondrocytes of rat intervertebral discs

作　　者：牛凯[1,2] 赵永见[3,2] 张雷[1] 李晨光[3,2] 王拥军[3,2] 郑为超[1] 

机构地区：[1]安徽理工大学医学院,安徽淮南232001 [2]上海中医药大学脊柱病研究所,上海200032 [3]上海中医药大学附属龙华医院,上海200032

出　　处：《药学学报》2014年第8期1136-1142,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81273777,81102606,81072831);国家重点基础研究发展计划(973计划)资助项目(2010CB530400);教育部长江学者和创新团队发展计划(PCSIRT,IRT1270)

摘　　要：本文探讨了中药单体苦杏仁苷(amygdalin)联合羟基红花黄色素A(HSYA)对IL-1β诱导退变的大鼠椎间盘软骨终板细胞的影响,寻找可能的作用机制。从1月龄SD大鼠椎间盘中分离软骨终板并培养软骨终板细胞,经鉴定后选择第3代软骨终板细胞用于实验,随机分为正常组、诱导组、苦杏仁苷组、羟基红花黄色素A组和联合用药组。采用CCK-8法检测细胞增殖,流式检测细胞凋亡,Real-time PCR(RT-PCR)检测Aggrecan、Col 2α1、Col 10α1、MMP-13、IL-1βmRNA的表达,细胞免疫荧光检测II型胶原、X型胶原的表达。结果表明,与正常组比较,IL-1β诱导组软骨终板细胞增殖降低,细胞凋亡增加(P<0.05),下调Aggrecan、Col 2α1 mRNA的表达,上调Col 10α1、MMP-13、IL-1βmRNA的表达(P<0.05);同时II型胶原蛋白的表达降低、X型胶原蛋白的表达升高。与IL-1β诱导组比较,苦杏仁苷组、羟基红花黄色素A组和联合用药组均能在抑制软骨终板细胞凋亡的同时促进软骨终板细胞的增殖(P<0.05),上调Aggrecan、Col 2α1 mRNA的表达的同时下调Col 10α1、MMP-13、IL-1βmRNA的表达(P<0.05);促进II型胶原蛋白的表达增强、X型胶原蛋白的表达降低。联合用药组作用的效果明显优于苦杏仁苷组、羟基红花黄色素A组(P<0.05)。苦杏仁苷联合羟基红花黄色素A可抑制IL-1β诱导的大鼠椎间盘软骨终板软骨细胞发生退变,优于苦杏仁苷、羟基红花黄色素A单独用药。The effect of amygdalin joint hydroxysaffior yellow A （HSYA） on the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1β and the possible mechanism were studied and explored. Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary endplate chondrocytes. After identification, they were divided into normal group, induced group, amygdalin group, HSYA group and combined group. CCK-8 kit was adopted to detect the proliferation of the endplate chondrocytes. FCM was measured to detect the apoptosis. Real-time PCR method was adopted to observe themRNA expression of Aggrecan, Col 2α1, Col 10 α1, MMP-13 and the inflammatory cytokines IL-1β. The protein expression of Col II, Col X was tested through immunofluorescence. Compared with the normal group, the proliferation of the endplate chondrocytes decreased while the apoptosis increased （P〈0.05）. With down regulation of the mRNA expressions of Aggrecan, Col 2 al and up regulation of the mRNA expressions of Col 10 α1, MMP- 13, IL-1β （P〈 0.05）, the protein expression of Col II decreased while the protein expression of Col X increased. Compared with the induced group, amygdalin group, HSYA group, the combined group could inhibit the apoptosis and promote the proliferation （P〈0.05）. They could increase the mRNA expressions of Aggrecan and Col 2 α1 while decrease the mRNA expressions of Col 10 α1, MMP-13 and IL-1β （P〈0.05）. They could also enhance the protein expression of Col II while reduce the protein expression of Col X. The effect of the combined group was significantly better than that of amygdalin and HSYA. Amygdalin joint HSYA could inhibit the degeneration of the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1β and better than the single use of amygdalin or HSYA.

关 键 词：苦杏仁苷 羟基红花黄色素A 椎间盘 退变 软骨终板 

分 类 号：R285[医药卫生—中药学]