香豆雌酚对膝骨关节炎患者软骨细胞OPG/RANKL及MMP-1、MMP-3表达的影响  

Effect of coumestrol on the expression of OPG / RANKL,MMP-1 and MMP-3 in the chondrocytes of patients with knee osteoarthritis

作　　者：王斌[1] 高益[1] 彭立波[1] 赵洪[1] 瞿玉兴[1] 

机构地区：[1]南京中医药大学附属常州市中医医院骨科,江苏常州213003

出　　处：《东南大学学报（医学版）》2015年第1期47-51,共5页Journal of Southeast University(Medical Science Edition)

基　　金：常州市科技计划项目(CY20130014)

摘　　要：目的:研究香豆雌酚(coumestrol)对膝骨关节炎患者软骨细胞OPG/RANKL及MMP-1、MMP-3表达的影响,初步探讨其对骨关节炎的治疗机制。方法:分离、培养因膝骨关节炎行膝关节置换患者的膝关节软骨细胞,运用Ⅱ型胶原免疫荧光染色进行软骨细胞鉴定。用不同浓度(10-9、10-8、10-7mol·L-1)香豆雌酚干预第3代细胞,并设0浓度为空白对照组,分别于培养后3、7 d用real-time PCR法测定细胞骨保护素(osteoprotegerin,OPG)、核因子κB受体活化因子配基(receptor activator of nuclear factor-κB ligand,RANKL)、基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)和基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)mRNA的表达。结果:干预3 d,10-8mol·L-1组OPG及RANKL mRNA表达均增加,其OPG/RANKL相对表达是空白对照组的2.59倍(P<0.05);干预7 d,10-9和10-8mol·L-1组OPG mRNA表达均增加,10-7mol·L-1组RANKL mRNA表达增加,10-8mol·L-1组OPG/RANKL相对表达是空白对照组的3.85倍(P<0.05)。干预3 d,10-8mol·L-1组MMP-1 mRNA表达减少(P<0.05);干预7 d,10-8mol·L-1组MMP-1和MMP-3 mRNA表达均减少(P<0.05)。结论:香豆雌酚能增加膝骨关节炎患者关节软骨细胞OPG表达及OPG/RANKL值,减少其MMP-1、MMP-3表达,具有一定的软骨保护作用。Objective：to study effect of coumestrol on the expression of OPG, RANKL, MMP-1 and MMP-3 in＆nbsp;the chondrocytes of patients with knee osteoarthritis（KOA）. Methods： Chondrocytes from KOA patients who had taken a total knee arthroplasty（ TKA） were isolated and cultured, and cells were identified with collagen typeⅡimmunofluorescence staining. The third generation cells were incubated in presence of various concentrations （10 -9 ,10 -8 ,10 -7 mol·L-1 ） of coumestrol, or 0 mol·L -1 for negtive control. real-time PCR assay was used to determine the OPG, RANKL, MMP-1 and MMP-3 mRNA expression level of the cells at the 3th and 7th day. Results：OPG/RANKL expression： At the 3th day, 10 -8 mol·L-1 coumestrol significantly increased OPG and RANKL mRNA expression and the ratio of OPG/RANKL gene expression was 2 . 59 times as much as that of the negtive control（P 〈0. 05）. At the 7th day, 10 -9 and 10 -8 mol·L-1 coumestrol significantly increased OPG mRNA expression, while 10 -7 mol · L-1 coumestrol significantly increased RANKL expression. And, in 10 -8 mol·L-1 coumestrol group, the ratio of OPG/RANKL gene expression was 3. 85 times as much as that of the negtive control（P〈0. 05）. MMP-1, MMP-3 expression：At the 3th day, 10 -8 mol·L-1 coumestrol significantly decreased MMP-1 mRNA expression（P 〈0. 05）. At the 7th day, 10 -8 mol·L-1 coumestrol significantly decreased MMP-1 and MMP-3 mRNA expression（P〈0. 05）. Conclusion：Coumestrol has its protective effects on cartilage by increasing the OPG mRNA expression and the ratio of OPG/RANKL gene expression and decreasing the MMP-1 and MMP-3 expression in the chondrocytes of patients with knee osteoarthritis.

关 键 词：香豆雌酚 软骨细胞 膝骨关节炎 骨保护素 基质金属蛋白酶 

分 类 号：R684.3[医药卫生—骨科学] R966[医药卫生—外科学]