TFDG对IL-1β体外诱导大鼠软骨细胞炎性损伤的保护作用研究

茶叶科学 ›› 2017, Vol. 37 ›› Issue (3): 290-298.

TFDG对IL-1β体外诱导大鼠软骨细胞炎性损伤的保护作用研究

周盈¹, 黄倩², 张甜¹, 陈萍^1,*

1. 浙江大学茶学系,浙江杭州 310058;
2. 浙江大学基础医学院,浙江杭州 310058

收稿日期:2016-10-12 修回日期:2017-01-05 出版日期:2017-06-15 发布日期:2019-08-22
通讯作者: *pingchen@zju.edu.cn
作者简介:周盈,女,硕士研究生,主要从事茶叶药理功能和茶叶品质鉴定研究,vaelailai@zju.edu.cn。
基金资助:
国家茶叶产业体系（CARS-23）、国家自然科学基金（31200521）、高等学校博士学科点专项科研基金（20120101120112）

Research on the Protective Effects of TFDG on IL-1β-induced Inflammatory Injury in Rat Chondrocytes in Vitro

ZHOU Ying¹, HUANG Qian², ZHANG Tian¹, CHEN Ping^1,*

1. Department of Tea Science, Zhejiang University, Hangzhou 310058, China;
2. School of Basic Medical Sciences, Zhejiang University, Hangzhou 310058, China

Received:2016-10-12 Revised:2017-01-05 Online:2017-06-15 Published:2019-08-22

摘要/Abstract

摘要： 体外分离培养SD雄性大鼠膝关节软骨细胞,经甲苯胺蓝及II型胶原（Col II）免疫荧光染色鉴定后,加入白介素-1β（IL-1β）诱导建立骨关节炎（OA）细胞模型,探讨茶黄素双没食子酸酯（TFDG）对OA软骨细胞的保护作用。细胞形态观察发现TFDG能明显改善OA软骨细胞形态。实时荧光定量PCR（Real-time PCR）结果显示,TFDG不仅可以上调软骨细胞分子标志物Col II mRNA的表达,还可以下调炎症因子IL-1β、IL-6 mRNA的表达。酶联免疫吸附实验（ELISA）检测结果进一步表明TFDG可明显降低炎症因子的分泌。免疫印迹（Western blot）检测结果证明,TFDG预干扰可降低炎症诱导酶环氧化酶COX-2蛋白表达量。这些结果说明TFDG通过减弱炎症反应,从而对IL-1β体外诱导大鼠软骨细胞炎性损伤起到保护作用。

关键词: 茶黄素双没食子酸酯, 骨关节炎, 软骨细胞, 炎症

Abstract: The chondrocytes were isolated and harvested from knee articular surface of male Sprague-Dawley rats, which were identified by toluidine blue and type II collagen immunofluorescence staining. To investigate potential protective effects of theaflavin-3,3′-digallate (TFDG) on IL-1β-induced inflammatory injury in rat chondrocytes in vitro, a model of osteoarthritis was established through stimulating rat chondrocytes with IL-1β. Inverted phase contrast microscopy showed TFDG treatment significantly improved osteoarthritis chondrocytes morphology. Real-time PCR results showed that TFDG treatment not only up-regulated chondrocyte marker Col II mRNA expression, but also down-regulated pro-inflammatory cytokines IL-1β and IL-6 mRNA expression. ELISA analysis further confirmed TFDG treatment significantly decreased the secretion of inflammatory factors. Western blot results showed that TFDG treatment significantly inhibited the inflammation-related enzyme COX-2 expression. Taken together, these results indicate that TFDG has a preventive effect on IL-1β-induced inflammatory injury in rat chondrocytes in vitro by reducing inflammatory reaction.

Key words: TFDG, osteoarthritis, chondrocytes, inflammation

中图分类号:

周盈, 黄倩, 张甜, 陈萍. TFDG对IL-1β体外诱导大鼠软骨细胞炎性损伤的保护作用研究[J]. 茶叶科学, 2017, 37(3): 290-298.

ZHOU Ying, HUANG Qian, ZHANG Tian, CHEN Ping. Research on the Protective Effects of TFDG on IL-1β-induced Inflammatory Injury in Rat Chondrocytes in Vitro[J]. Journal of Tea Science, 2017, 37(3): 290-298.

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