白藜芦醇对软骨细胞去分化现象的作用机制

doi:10.16333/j.1001-6880.2017.1.002

天然产物研究与开发 ›› 2017, Vol. 29 ›› Issue (1): 7-13.doi: 10.16333/j.1001-6880.2017.1.002

白藜芦醇对软骨细胞去分化现象的作用机制

陈文杰1,2，耿书国3，殷嫦嫦4，李忠友1,2，殷明1*

1 南昌大学第二附属医院骨科；2 南昌大学研究生院医学部，南昌 330006；3 九江市武宁县人民医院，九江 332300；4 九江学院九江市转化医学重点实验室，九江 332000

出版日期:2017-01-31 发布日期:2017-02-17

Possible Mechanism of Resveratrol on Dedifferentiation of Chondrocyte

CHEN Wen-jie1,2,GENG Shu-guo3, YIN Chang-chang4,LI Zhong-you1,2, YIN Ming1*

1 Department of orthopedics,The Second Hospital Affiliated of Nanchang University; 2 Medicine School of Graduate Center,Nanchang University,Nanchang 330006,China; 3 Wuning County People’s Hospital,Jiujiang 332300,China; 4 Jiujiang Key Laboratory of Translational Medicine,Jiujiang College,Jiujiang 332000,China

Online:2017-01-31 Published:2017-02-17

摘要/Abstract

摘要： 该实验观察白藜芦醇（Resveratrol，Resv）对兔关节软骨细胞去分化现象的作用并探讨其可能的机制。首先，在无菌条件下取4周龄兔关节软骨细胞，行软骨细胞鉴定后将细胞随机分成5组：空白组、0 μmol/L Resv组、2.5μmol/L Resv组、5 μmol/L Resv组和10 μmol/L Resv，体外单层培养传代至第5代。分别取P1、P3、P5代细胞在倒置相差显微镜下观察细胞形态变化，倒置相差显微镜下观察到各组P1代软骨细胞形态基本一致，P3、P5代2.5 μmol/L Resv组细胞形态变化不大，其余各组细胞出现胞体变大，胞浆变淡，镜下胞核不明显等衰老的形态变化；Alcian Blue染色检测蛋白聚糖（Aggrecan）含量和β-半乳糖苷酶染色检测衰老软骨细胞，各组P1代细胞Alcian Blue染色和β-半乳糖苷酶染色无明显差别，P3、P5代2.5 μmol/L白藜芦醇组较其他组明显深染、衰老软骨细胞数明显减少；RT-PCR检测蛋白多糖（Aggrecan），沉默信息调节因子1（SIRT1），抑癌基因p53、p21以及P16的表达，2.5μmol/L Resv组的Aggrecan和SIRT1基因表达明显上调，P53、P21及P16基因表达显著下调。综上所述：低浓度白藜芦醇（2.5 μmol/L）可能通过上调SIRT1，下调P53、P21、P16基因的表达抑制软骨细胞去分化的现象。

关键词: 白藜芦醇, 软骨细胞, SIRT1, 衰老, 去分化

Abstract: In this study,the role of resveratrol on dedifferentiation of rat chondrocyte and its possible mechanism were investigated.Cartilage cells was harvested under sterile conditions from normal rabbit′s joint of 4 weeks old rabbit and was then identified.The experiment were randomly divided into five groups:control group,0 μmol/L Resv group,2.5 μmol/L Resv group,5 μmol/L Resv group and 10 μmol/L group.The rabbit articular chondrocytes were subcultured in vitro to the 5th generation (named P1-P5).Inverted microscope was used to observe cell morphology at 1,3 and 5 generation respectively.The results indicated that the morphology at passage 1 chondrocytes were similar.The morphology of 2.5 μmol/L Resv group at passage 3 and 5 were nearly the same with passage 1,but other groups became enlarged,cytoplasmic stained light and the nucleus were not obvious;Alcian Blue staining was used to observe the changes of Proteoglycans.The results of Alcian Blue staining showed that the poly polysaccharide protein expression of 2.5 μmol/L Resv group were higher than other groups; β-galactosidase staining were used to identify the ageing cells.The results showed that the number of aging cells of 2.5 μmol/L Resv group was lower than other groups.RT-PCR were adopted to detect the RNA expression level of Aggrecan gene,SIRT1 gene,p53 gene,p21gene and P16 gene.RT-PCR analysis showed that SIRT1 gene of2.5 μmol/L Resv was significantly upregulated,and P53,P21 and P16 gene was downregulated.In conclusion,this experiment showed that 2.5 μmol/L Resv can inhibit the dedifferentiation of chondrocytes via upregulating expression of SIRT1 gene and downregulating expression of P53,P21 and P16.

Key words: resveratrol, chondrocyte, SIRT1, aging, dedifferentiation

中图分类号:

R681.3

陈文杰1,2，耿书国3，殷嫦嫦4，李忠友1,2，殷明1*. 白藜芦醇对软骨细胞去分化现象的作用机制[J]. 天然产物研究与开发, 2017, 29(1): 7-13.

CHEN Wen-jie1,2,GENG Shu-guo3, YIN Chang-chang4,LI Zhong-you1,2, YIN Ming1*. Possible Mechanism of Resveratrol on Dedifferentiation of Chondrocyte[J]. NATURAL PRODUCT RESEARCH AND DEVELOPMENT, 2017, 29(1): 7-13.

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白藜芦醇对软骨细胞去分化现象的作用机制

Possible Mechanism of Resveratrol on Dedifferentiation of Chondrocyte

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