[1]胡鸿扬 薛艳 曹月龙 庞坚 郑昱新 丁道芳△.反式维甲酸诱导大鼠关节软骨细胞的退变[J].中国中医骨伤科杂志,2018,26(08):1-4，10.
　HU Hongyang XUE Yan CAO Yuelong PANG Jian ZHENG Yuxin DING Daofang.Establishment of Articular Chondrocyte Degeneration of Rats by Trans Retinoic Acid[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2018,26(08):1-4，10.
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反式维甲酸诱导大鼠关节软骨细胞的退变()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第26卷
期数:
2018年08期
页码:
1-4，10
栏目:
实验研究
出版日期:
2018-08-02

文章信息/Info

Title:
Establishment of Articular Chondrocyte Degeneration of Rats by Trans Retinoic Acid
文章编号:
1005-0205(2018)08-0001-04
作者:
胡鸿扬1; 2 薛艳1; 2 曹月龙1; 2 庞坚1; 2 郑昱新1; 2 丁道芳1; 2△
1上海中医药大学附属曙光医院石氏伤科医学中心 (上海,201203) 2上海市中医药研究院骨伤科研究所
Author(s):
HU Hongyang1; 2 XUE Yan1; 2 CAO Yuelong1; 2 PANG Jian1; 2 ZHENG Yuxin1; 2 DING Daofang1; 2△
1Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 2Institute of Traumatology & Orthopedics, Shanghai Academy of Traditional Chinese Medicine, Shanghai 201203, China.
关键词:
反式维甲酸;  细胞增殖;  软骨退变;  细胞形态
Keywords:
Keywords: trans retinoic acid(TRA);  cell proliferation;  chondrocytes' degeneration;  cell morphology
分类号:
R-33
文献标志码:
A
摘要:
目的:观察反式维甲酸(TRA)对体外培养大鼠软骨细胞的生长增殖、形态和软骨退变相关基因表达的影响。方法:取出生24 h SD大鼠关节处软骨,Ⅱ型胶原酶多次消化后获得原代软骨细胞,取P1代细胞进行实验。对照组软骨细胞于体积分数为10%的胎牛血清(FBS)的高糖DMEM培养基中进行培养,而TRA组是在对照组培养基的基础上加入10 μmolTRA进行培养。培养24 h后,观察细胞形态变化,MTT法检测两组细胞的增殖情况,Western Blot检测Ⅱ型胶原和MMP13的表达,DMMB法测定细胞中GAG的含量,定量PCR检测ADAMTS5,MMP9,A-can,Sox9,COLX和ALP的表达。结果:TRA组的软骨形态由原来的多角形变成长梭形,且TRA组抑制软骨细胞的增殖,同时下调基质蛋白Ⅱ型胶原,A-can,GAG含量及转录因子Sox9表达,上调MMP13,MMP9,ADAMTS5,COLX及ALP的表达。结论:TRA具有促进软骨退变的作用,可用于建立体外软骨退变模型。
Abstract:
Abstracts Objective:To investigate the effects of trans retinoic acid(TRA)on the chondrocyte' proliferation, morphology, and the expression levels of cartilage degeneration associated gene in rat chondrocytes in vitro.Methods: Chondrocyte was cultured from articular cartilages of newly born(within 24 h)SD rats, and digested with type Ⅱ collagenase.Chondrocyte at passage one was used for the experiment.The chondrocyte was cultured with H-DMEM medium containing 10% FBS in Control group, and H-DMEM medium containing 10% FBS plus 10 μmol trans retinoic acid in TRA group.The morphology change was investigated under phase contrast microscope after chondrocyte were treated with trans retinoic acid for 24 h.Chondrocytes' proliferation was detected by MTT method, and the protein expression levels of type Ⅱ collagen and MMP13 were examined by Western-blot.The GAG contents in chondrocytes were detected by DMMB method.The mRNA levels of ADAMTS5, MMP9, A-can, Sox9, ALP and ColX were detected by real-time PCR.Results: The cell morphology was changed from polygon to fibro-like chondrocytes in TRA group compared to control group, and the proliferation of chondrocytes in TRA group were inhibited.The results showed that the expression levels of type Ⅱ collagen, A-can, GAG and Sox9 were decreased while the expression levels of MMP13, MMP9, ADMATS5, COLX and ALP were up-regulated in TRA group compared with control group.Conclusion: It was proved that TRA could be used as an efficient inducer in establishing OA model in vitro which induces chondrocytes' degeneration.

参考文献/References:

[1] Kroman SL,Roos EM,Bennell KL,et al.Measurement properties of performance-based outcome measures to assess physical function in young and middle-aged people known to be at high risk of hip and/or knee osteoarthritis:a systematic review[J].Osteoarthritis Cartilage,2014,22(1):26-39. [2] Corbett MS,Rice SJC,Madurasinghe V,et al.Acupuncture and other physical treatments for the relief of pain due to osteoarthritis of the knee:network meta-analysis[J].Centre for Reviews and Dissemination(UK),2013,21(9):1290-1298. [3] 张宇,赵永见,周泉,等.IL-1β诱导小鼠椎间盘软骨终板细胞炎性退变的细胞模型研究[J].免疫学杂志,2012,28(8):665-668. [4] Zhang Y,Ross AC.Retinoic Acid and the transcription factor mafB act together[J].J Cell Biochem,2013,114(2):471-479. [5] Suri S,Gill SE,Massena de Camin S,et al.Neurovascular invasion at the osteochondral junction and in osteophytes in osteoarthritis[J].Ann Rheum Dis,2007,66(11):1423-1428. [6] Aigner T,Sachse A,Gebhard PM,et al.Osteoarthritis:pathobiology-targets and ways for therapeutic intervention[J].Adv Drug Deliv Rev,2006,58(2):128-149. [7] Goldring MB.Update on the biology of the chondrocyte and new approaches to treating cartilage diseases[J].Best Pract Res Clin Rheumatol,2006,20(5):1003-1025. [8] Troeberg L,Nagase H.Proteases involved in cartilage matrix degradation in osteoarthritis[J].Biochim Biophys Acta,2012,1824(1):133-145. [9] Lin EA,Liu CJ.The role of ADAMTS in arthritis [J].Protein,2010,1(1):33-47. [10] Wu CW,Tchetina EV,Mwale F,et al.Proteolysis involving matrix metalloproteinase 13(collagenase-3)is required for chondrocyte differentiation that is associated with matrix mineralization[J].J Bone Miner Res,2002,17(4):639-651. [11] Dahlberg L,Billinghurst RC,Manner P,et al.Selective enhancement of collagenase-mediated cleavage of resident type Ⅱ collagen in cultured osteoarthritic cartilage and arrest with a synthetic inhibitor that spares collagenase 1(matrix metalloproteinase 1)[J].Arthritis Rheum,2000,43(3):673-682. [12] Aigner T,Dudhia J.Phenotypic modulation of chondrocytes as a potential therapeutic target in osteoarthritis:a hypothesis[J].Ann Rheum Dis,1997,56(5):287-291. [13] Sekiya I,Tsuji K,Koopman P,et al.SOX9 enhances aggrecan gene promoter/enhancer activity and is up-regulated by retinoic acid in a cartilage-derived cell line,TC6[J].J Biol Chem,2000,275(15):10738-10744. [14] Ushijima T,Okazaki K,Tsushima H,et al.CCAAT/enhancer-binding protein β regulates the repression of type Ⅱ collagen expression during the differentiation from proliferative to hypertrophic chondrocytes[J].J Biol Chem,2014,289(5):2852-2863. [15] Okubo Y,Reddi AH.Thyroxine downregulates Sox9 and promotes chondrocyte hypertrophy [J].Biochem Biophys Res Commun,2003,306(1):186-190. [16] Song Y,Xu K,Yu C,et al.The use of mechano growth factor to prevent cartilage degeneration in knee osteoarthritis[J].J Tissue Eng Regen Med,2018,12(3):738-749. [17] 李保驰,王维山,董金波,等.MMP-3、MMP-13在骨性关节炎患者滑膜中的表达及意义[J].中国骨质疏松杂志,2014,20(6):593-596.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目 (81073114,81072830,81503598) △通信作者 E-mail:d.wilhel@qq.com
更新日期/Last Update: 2018-08-02