[1]杜国庆 丁道芳 李玲慧 宋奕 王辉昊 张玉民 郑昱新 詹红生△.不同浓度的Chir99021对软骨细胞增殖和Ⅱ型胶原表达的影响[J].中国中医骨伤科杂志,2016,24(02):1-5.
　DU Guoqing DING Daofang LI Linghui SONG Yi WANG Huihao ZHANG Yumin ZHENG Yuxin ZHAN Hongsheng.Effect of Different Concentrations of Chir99021 on Proliferation and Col2a1 Expression of Neonatal Rat Chondrocytes[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2016,24(02):1-5.
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不同浓度的Chir99021对软骨细胞增殖和Ⅱ型胶原表达的影响
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第24卷
期数:
2016年02期
页码:
1-5
栏目:
实验研究
出版日期:
2016-02-14

文章信息/Info

Title:
Effect of Different Concentrations of Chir99021 on Proliferation and Col2a1 Expression of Neonatal Rat Chondrocytes
文章编号:
1005-0205(2016)02-0001-05
作者:
杜国庆1; 2 丁道芳1; 2 李玲慧1; 2 宋奕3 王辉昊1; 2 张玉民1; 2 郑昱新1; 2 詹红生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
3.上海中医药大学
△.通信作者 E-mail:zhanhongsheng2010@163.com
Author(s):
DU Guoqing1; 2 DING Daofang1; 2 LI Linghui1; 2 SONG Yi3 WANG Huihao1; 2 ZHANG Yumin1; 2 ZHENG Yuxin1; 2 ZHAN Hongsheng1; 2△
1.Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203,China;
2.Institute of Traumatology & Orthopedics, Shanghai Academy of Traditional Chinese Medicine, Shanghai 201203,China;
3.Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China.
关键词:
Wnt/β-catenin;  软骨细胞;  增殖;  Ⅱ型胶原
Keywords:
Wnt/β-catenin;  chondrocyte;  proliferation;  typeⅡcollagen
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨不同浓度的Wnt/β-catenin信号通路激活剂Chir99021对体外培养的大鼠软骨细胞增殖和Ⅱ型胶原表达的影响。方法:新生24 h SD大鼠8只(雌雄不限),取出关节软骨进行软骨细胞培养,细胞培养48 h后进行软骨细胞免疫荧光鉴定。以不同药物浓度将软骨细胞分为四组,分别加入浓度为0.00 μmol/L(对照组),0.75 μmol/L,1.50 μmol/L,3.00 μmol/L的Wnt/β-catenin信号通路激活剂Chir99021进行干预。24 h后分别观察不同浓度组细胞形态的变化,蛋白印迹分析法(Western blot)检测Ⅱ型胶原、β-catenin和PCNA蛋白的表达情况。结果:在0.00 μmol/L(对照组),0.75 μmol/L,1.50 μmol/L,3.00 μmol/L的Wnt/β-catenin信号通路激活剂Chir99021干预下,各组细胞数量明显逐渐增加,与对照组相比较,各加药组β-catenin和PCNA蛋白的表达逐渐上调(P<0.05),而Ⅱ型胶原蛋白的表达逐渐下降(P<0.05)。结论:不同浓度的Chir99021激活Wnt/β-catenin信号通路后明显促进了软骨细胞的增殖,却抑制了软骨细胞分泌的Ⅱ型胶原。
Abstract:
Objective:To investigate the effect of different concentrations of chir99021 on the proliferation and the expression of typeⅡcollagen(Col2a1)of rat chondrocytes. Methods: Eight 24 h old SD rats(female or male)were executed by dislocating. The articular cartilage of rats were removed and cuted into blocks of 1 mm×1 mm size. The articular cartilage were digested by type Ⅱcollagenase for four minutes five times. The mixed cells were cultured in thermostat incubator with 5%CO2 under the condition of 37 ℃. Chondrocytes were identificatied by the immunofluorescence. Rat chondrocytes were cultured with different concentrations of chir99021 for 24 h. The concentrations were 0.00 μmol/L, 0.75 μmol/L, 1.50 μmol/L, 3.00 μmol/L. Morphological changes, cell proliferation and protein expression levels of PCNA, β-catenin and Col2a1 were investigated and compared among four groups. Results: The chir99021 obviously promoted the proliferation of chondrocytes of newly born(within 24 h)SD rats and the promotion rate was associated with the concentration of chir99021. Compared with control group, the expression levels of both PCNA and β-catenin were upregulated while the expression level of Col2a1 was downregulated with the increasing concentration of chir99021(P<0.05). Conclusion: The different concentrations of chir99021 obviously promoted the proliferation of chondrocytes and inhibited the expression of typeⅡcollagen through activating Wnt/β-catenin signaling.

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备注/Memo

备注/Memo:
收稿日期:2015-05-09
基金项目:“中医骨伤科学”国家重点学科项目(100508)
上海市中医药领军人才建设项目(2012-63-15)
上海领军人才项目(041)
“海派中医流派传承研究基地”项目(ZYSNXD- CC-HPGC-JD-001)
国家自然科学基金项目(81173277,81072830)
上海市科委重点科技攻关项目(11DZ1972902, 11DZ1973402)
上海市中医药事业发展三年行动计划项目 (ZYSNXD-CC-ZDYJ047)
上海市卫生局中药新药及院内制剂研发项目 (2011ZJ007,2011ZJ012)
上海市教育委员会科研创新项目(11YZ64, 12YZ064)
更新日期/Last Update: 2016-02-02