苦杏仁苷对IL-1β诱导后大鼠椎间盘软骨终板细胞的影响  

Effects of amygdalin on the endplate chondrocytes induced by IL-1β derived from rats' intervertebral discs

作　　者：牛凯[1,2] 赵永见[3,2] 姚长风[2] 梁倩倩[3,2] 李晨光[3,2] 张宇[1] 郑为超[1] 王拥军[3,2] 

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：《中国药理学通报》2013年第12期1725-1730,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81273777,81072831,81102605,81102606)

摘　　要：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,随机分组为正常组、诱导组、苦杏仁苷10-2、10-3、10-4、10-5mol·L-1给药组,CCK-8法检测各组对大鼠椎间盘软骨终板细胞增殖的影响。Real-time PCR(RTPCR)检测聚集蛋白聚糖(aggrecan-1)、Ⅱ型胶原(type II collagen,Col2a1)、Ⅹ型胶原(type X collagen,Col10al)、基质金属蛋白酶-13(matrix metalloproteinase 13,MMP-13)基因的表达情况。细胞免疫荧光检测分析Col2a1、Col10al的表达,流式细胞仪检测其对软骨终板细胞凋亡的影响。结果苦杏仁苷10-2mol·L-1给药组能够抑制椎间盘软骨终板细胞的增殖,与正常组比较差异有统计学意义(P<0.05)。流式分析,其细胞凋亡比例较诱导组及正常组均较高。RT-PCR检测结果显示一定浓度的苦杏仁苷能够上调Aggrecan、Col2a1mRNA的表达,下调Col10al、MMP-13 mRNA的表达,与诱导组比较,差异有统计学意义(P<0.05)。一定浓度的苦杏仁苷能够上调Col2a1蛋白的表达,下调Col10al蛋白的表达。结论一定浓度的苦杏仁苷能够抑制IL-1β诱导大鼠椎间盘软骨终板细胞发生退变,起到延缓椎间盘退变的作用。Aims To observe the effect of different concentrations of amygdalin on the endplate chondro- cytes induced by IL-1β derived from rats＇ interverte- bral discs and to further explore the possible mecha- nism. Methods Chondrocytes were obtained from endplate of one-month SD rats＇ intervertebral discs and primary chondrocytes were cultured. After identifying, they were divided into normal group, induced group and amygdalin 10-2, 10 3 10-4, 10-5 mol · L^-1 ad- ministration groups. CCK-8 kit was used to measure the proliferation of the endplate ehondrocytes. Real- Time PCR was adopted to detect the mRNA expression of Aggreean, Col2al, Coll0al, MMP-13. The protein expression of Col2al , Colloal was stained by immuno- fluorescence and the apoptosis was detected by flow cy- tometry （FCM）. Results Amygdalin 10-2 mol · L-1 inhibited the proliferation of the endplate chondroeytesand increased the apoptotic rate. Compared with the normal group the differences were statistically signifi- cant （P 〈0. 05）. RT-PCR showed that a certain con- centration of amygdalin increased the mRNA expression of Aggrecan, Col2al and decreased the mRNA expres- sion of Coll0al, MMP-13. The mRNA expression differences were statistically significant （ P 〈 0. 05 ） be- tween normal group and the group of amygdalin with different concentrations. A certain concentration of a- mygdalin increased the protein expression of Col2al and reduced the protein expression of Coll0al. Con- clusions A certain concentration of amygdalin can in- hibit the degeneration of the endplate chondrocytes in- duced by IL-1β derived from rats＇ intervertebral discs and improve the degeneration of intervertebral discs.

关 键 词：苦杏仁苷 椎间盘 退变 软骨终板 IL-1Β 凋亡 

分 类 号：R-332[医药卫生] R323.4