来曲唑抑制大鼠生长板软骨细胞凋亡的最适药物浓度筛选  

Screening of the Optimal Drug Concentration of Letrozole for Inhibiting Apoptosis of Rat Growth Plate Chondrocytes

作　　者：王朋涛 谢艳 杨艳梅 孔凡国 王娜 张海龙 WANG Pengtao;XIE Yan;YANG Yanmei;KONG Fanguo;WANG Na;ZHANG Hailong(Henan University of Traditional Chinese Medicine, Henan Zhengzhou 450046,China;Henan Provincial Luoyang Orthopedic-Traumatological Hospital/Henan Provincial Orthopedic Hospital,Henan Luoyang 471002,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南省洛阳正骨医院/河南省骨科医院,河南洛阳471002

出　　处：《中国医药导刊》2021年第8期611-616,共6页Chinese Journal of Medicinal Guide

基　　金：河南省中医药科学研究专项课题(课题编号:2018ZY1023,课题名称:延迟骨骺闭合的芳香化酶抑制剂药物筛选)。

摘　　要：目的:筛选来曲唑(letrozole,LE)抑制大鼠生长板软骨细胞凋亡的最适药物浓度。方法:解剖取大鼠胫骨及股骨生长板软骨,采用二步酶消法体外分离获得生长板软骨细胞并进行培养,分别利用倒置相差显微镜及Ⅱ型胶原免疫荧光实验对生长板软骨细胞进行形态学观察和鉴定。用IL^(-1)β诱发生长板软骨细胞凋亡,分别加入药物浓度为2.5、5、10、15、20μmol·L^(-1)来曲唑的培养基进行培养,流式细胞仪检测来曲唑抑制细胞凋亡情况。同样以IL^(-1)β诱发生长板软骨细胞凋亡,分别加入药物浓度为1、2.5、5、10、15μmol·L^(-1)来曲唑的培养基进行培养,MTT法筛选来曲唑抑制细胞凋亡的最适药物浓度。结果:来曲唑浓度为2.5、5、10、15μmol·L^(-1)时均可以不同程度抑制生长板软骨细胞凋亡。模型对照组细胞OD值较正常对照组降低(P<0.01)。DMSO组与非造模组与正常对照组细胞OD值差异无统计学意义。结论:药物浓度为2.5μmol·L^(-1)的来曲唑抑制生长板软骨细胞凋亡效果最好,可以做为今后其他抑制细胞凋亡药物的阳性对照药物。Objective:To screen the optimal concentration of letrozole(LE)to inhibit the apoptosis of rat growth plate chondrocytes.Methods:Rat tibial and femoral growth plate cartilage were dissected,and growth plate chondrocytes were isolated and cultured by two-step enzyme elimination method.The morphology of growth plate chondrocytes was observed and identified by inverted phase contrast microscope and typeⅡcollagen immunofluorescence test.The chondrocyte apoptosis of growth plate was induced by IL^(-1)β,and the letrozole concentrations of 2.5,5,10,15 and 20μmol·L^(-1) respectively were added to culture the cells.The inhibition of letrozole on cell apoptosis was detected by flow cytometry.Also apoptosis chondrocytes in growth plate was induced by IL^(-1)β,the letrozole concentrations were 1,2.5,5,10 and 15μmol·L^(-1) respectively were added to culture the cells.The optimal concentration of letrozole to inhibit apoptosis was screened by MTT method.Results:The letrozole concentrations of 2.5,5,10 and 15μmol·L^(-1) could inhibit the apoptosis of growth plate chondrocytes in varying degrees.The OD value of model control group was lower than that of normal control group(P<0.01).There was no significant difference in cell OD between DMSO group,non model group and normal control group.Conclusion:The letrozole concentration of 2.5μmol·L^(-1) has the best effect on inhibiting apoptosis of growth plate chondrocytes,and can be used as a positive control drug for other anti-apoptosis drugs in the future.

关 键 词：来曲唑 生长板软骨细胞 凋亡 最适药物浓度 

分 类 号：R329.252[医药卫生—人体解剖和组织胚胎学]