摘要
采用白介素-1β(interleukin-1β,IL-1β)对体外培养的正常大鼠软骨细胞进行诱导,构建骨关节炎(osteoarthritis,OA)炎性退变细胞模型,探讨茶黄素(theaflavins,TFs)治疗骨关节炎的可行性。通过甲苯胺蓝及免疫荧光染色对细胞进行鉴定,利用CCK-8细胞活力检测筛选TFs药物浓度;取第2代生长状况良好的大鼠膝关节软骨细胞,根据所加培养物的不同将实验分为3组:空白组G0、模型组G1(10 ng/mL IL-1β刺激)、TFs组G2(不同浓度的TFs+10 ng/mL IL-1β刺激),通过实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测各组Ⅱ型胶原(typeⅡcollagen,ColⅡ)、蛋白聚糖(aggrecan,ACAN)、基质金属蛋白酶-13(matrix metalloproteinases-13,MMP-13)、IL-1β及环氧合酶-2(cyclooxygenase-2,COX-2)mRNA的表达。结果表明,甲苯胺蓝及免疫荧光染色结果呈阳性,证实体外分离培养的细胞为软骨细胞。CCK-8检测结果显示:TFs质量浓度在100μg/mL时,细胞活力较空白组明显减弱(P<0.05);TFs质量浓度在0~75μg/mL时不影响软骨细胞活力。qRT-PCR结果表明:与空白组相比,模型组中合成因子ColⅡ和ACAN mRNA表达量极显著降低(P<0.01),分解因子MMP-13、炎症细胞因子IL-1β及炎症诱导酶COX-2 mRNA表达量极显著增加(P<0.01);与模型组相比,TFs组能抑制IL-1β引起的炎症相关因子mRNA的表达,且呈浓度依赖性(P<0.05)。综上所述,茶黄素可通过增强软骨细胞合成因子活性、减弱分解因子活性并抑制细胞炎症反应,有效延缓大鼠软骨细胞炎性退变进程,对炎症软骨细胞有一定的保护作用。
Osteoarthritis(OA)is a degenerative joint disease with an obviously increasing morbidity as age increases,which seriously affects old people’s joint function and life quality.The key to pathological changes in OA is the damage and loss of articular cartilage.Inflammatory cytokines,such as interleukin-1β(IL-1β),could induce cartilage degeneration and inflammatory reaction through a series of cascade reactions,especially in cartilage lesions.Theaflavins(TFs)are the main active ingredients of black tea polyphenols with anti-inflammation and antioxidant functions.This study focuses on the protective effects of TFs against inflammatory degeneration of rat chondrocytes and evaluates the feasibility of TFs in the treatment of OA.
作者
周盈
黄倩
王帅
陈萍
ZHOU Ying;HUANG Qian;WANG Shuai;CHEN Ping(Department of Tea Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China;School of Basic Medical Sciences, Zhejiang University Hangzhou 310058, China)
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2017年第4期511-518,共8页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
国家茶叶产业技术体系项目(CARS-23)
