Tang Xiangyu,Cao Zhen,Dong Shiwu,et al.Effect of resveratrol on chondrocyte hypertrophy and related mechanism[J].J Third Mil Med Univ,2016,38(12):1392-1397.

白藜芦醇促进软骨细胞肥大化的作用机制研究(/HTML )




Effect of resveratrol on chondrocyte hypertrophy and related mechanism
Tang Xiangyu Cao Zhen Dong Shiwu Wu Jimin

Institute of Medical Equipment, Academy of Military Medical Sciences, Tianjin, 300161; Department of Biomedical Material Sciences, School of Biomedical Engineering, Third Military Medical University, Chongqing, 400038, China

pre-chondrocyte cells resveratrol hypertrophy endochondral ossification
R285.5; R322.71; R329.2

目的      探讨白藜芦醇(resveratrol,RES)对软骨前体细胞ATDC5肥大化的影响及其可能机制。      方法      通过CCK-8法研究不同浓度RES对ATDC5细胞活性的影响。建立软骨肥大化诱导体系,根据活性实验的安全浓度,设立对照组(0 μmol/L)和RES组(1 μmol/L)。qRT-PCR和Western blot法确定肥大化相关因子RUNX2、MMP13和COLX表达变化;甲苯胺蓝和免疫组化分析细胞外基质粘多糖及COLX、RUNX2表达;利用qRT-PCR检测软骨细胞肥大化经典通路WNT、IHH和FGF中相关因子的表达变化。      结果      RES安全使用浓度为1 μmol/L;与对照组相比,在肥大化诱导14 d后,RES组ATDC5细胞肥大化水平明显提升;RES组肥大化相关基因表达量在3、7、14 d时明显增加;WNT和IHH途径中促肥大化因子β-catenin和GLI2的表达水平在RES组较对照组升高,而GLI3表达水平下降。FGF途径中PI3K的表达水平较对照组没有变化。      结论      RES通过激活β-catenin及GLI2,抑制GLI3,促进RUNX2表达,实现促进ATDC5肥大化的目的。


Objective      To investigate the effect of resveratrol (RES) on hypertrophic differentiation of pre-chondrocyte cells (ATDC5) and related mechanism.       Methods      ATDC5 cells were treated with basal culture medium with different concentrations of RES for 24 h and 48 h, followed by CCK-8 assay. The hypertrophy inducing system was established. ATDC5 cells were divided into control group (0 μmol/L) and RES group (1 μmol/L). The expression levels of hypertrophy-related factors including RUNX2, MMP13 and collagens X (COLX) were detected using qRT-PCR and Western blotting. Toluidine blue staining for GAG and immunohistochemistry for COLX and RUNX2 were performed. Expression levels of altered transcriptional factors and key molecules in canonical chondrogenic pathways including WNT, IHH and FGF were examined by qRT-PCR.       Results      RES with concentration below 1 μmol/L had no toxic effect in this experiment. After induction with hypertrophy medium for 14 d, the hypertrophy of ATDC5 cells was increased after RES treatment. The treated ATDC5 cells showed a significant increase both at the mRNA and protein levels of hypertrophy-related genes including RUNX2, MMP13 and COLX on day 3, day 7, and day 14. The expression levels of β-catenin and GLI2 in the RES group were increased and that of  GLI3 was decreased significantly. Expression of PI3K was not changed in the RES group.       Conclusion      RES promotes hypertrophy of ATDC5 cells by up-regulating β-catenin and GLI2, down-regulating GLI3 and promoting expression of RUNX2.


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更新日期/Last Update: 2016-06-07