亚麻木酚素对大鼠生长板软骨细胞凋亡的影响

Effect of Flax lignans on apoptosis of growth plate chondrocytes in rats

目的:比较不同浓度亚麻木酚素抑制生长板软骨细胞凋亡的效果,筛选出最适药物浓度,为其延迟大鼠骨骺闭合促进长骨生长进行理论支持。方法:选取2只4周龄雄性SD大鼠,SPF级,体质量80 g。解剖大鼠胫骨及股骨生长板软骨并进行体外分离,获得生长板软骨细胞进行培养,利用倒置相差显微镜及Ⅱ型胶原免疫荧光实验对软骨细胞分别进行形态学观察、鉴定,然后采用白细胞介素-1β(interleukin-1β,IL-1β)20 ng/ml诱发生长板软骨细胞凋亡为模型组,同时加入分别含1、10、20、40μM亚麻木酚素为实验组,同时加入5μM来曲唑为阳性对照组,分别培养24、48 h,MTT法观察药物促进细胞增殖情况,流式细胞仪检测药物抑制细胞凋亡情况。结果:含量1、10、20、40μM的亚麻木酚素均可不同程度促进细胞增殖,其原理为药物抑制了IL-1β诱发的生长板软骨细胞凋亡,抑制细胞凋亡的最佳药物浓度为20μM。结论:适宜浓度的亚麻木酚素对大鼠生长板软骨细胞凋亡抑制效果明显且最适药物浓度为20μM,为骨骼延迟闭合提供更长的生长时间,从而为促进发育期骨骼增长提供可能。

Objective:To compare the effects of different concentrations of linolenin on inhibiting apoptosis of chondrocytes in the growth plate,and to screen the optimal concentration of linolenin,so as to provide theoretical support for delaying epiphyseal closure and promoting long bone growth in rats.Methods:Two 4-week-old male SD rats(SPF grade)with a body mass of 80 g were selected.The growth plate cartilage of rat tibia and femur was dissected and isolated in vitro to obtain growth plate chondrocytes for culture.The chondrocytes were observed and identified by inverted phase contrast microscope and typeⅡcollagen immunofluorescence test,and then 20 ng/ml IL-1βwas used to induce apoptosis of growth plate chondrocytes as model group,and added with 1,10,20,40μM linolenin as the experimental group,and 5μM letrozole as the positive control group.The cells were cultured for 24 and 48 hours respectively.The drug promoted cell proliferation was observed by MTT method,and the drug inhibited cell apoptosis was detected by flow cytometry.Results:Contents 1,10,20,40μM could promote cell proliferation in varying degrees,and the principle was that the drug inhibits IL-1βinduced chondrocyte apoptosis in the growth plate,and the optimal concentration of drugs to inhibit apoptosis was 20μM.Conclusion:The appropriate concentration of linseed lignans can significantly inhibit the apoptosis of chondrocytes in the growth plate of rats,and the optimal drug concentration is 20μM.It provides possibility for delayed bone closure and longer growth time to promote bone growth during development.