白藜芦醇对软骨细胞去分化现象的作用机制

发布时间：2018-08-18 10:59

【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、0μmol/L Resv组、2.5μmol/L Resv组、5μmol/L Resv组和10μmol/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2.5μmol/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2.5μmol/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(2.5μmol/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。
[Abstract]:The effect of resveratrol on dedifferentiation of rabbit articular chondrocytes was observed and its possible mechanism was discussed. Firstly, the articular chondrocytes of rabbits at 4 weeks of age were collected under aseptic condition. The chondrocytes were randomly divided into five groups: blank group (0 渭 mol/L Resv), 2.5 渭 mol/L Resv group (5 渭 mol/L Resv) and 10 渭 mol/L Resv group. The changes of cell morphology were observed under inverted phase contrast microscope. The morphology of P1 passage chondrocytes in each group was basically consistent with that of P3 / P5 passage 2.5 渭 mol/L Resv group. The morphologic changes of senescence such as light cytoplasm and unclear nucleus under microscope were detected by Alcian Blue staining and 尾 -galactosidase staining for the detection of the content of proteoglycan (Aggrecan) and 尾 -galactosidase staining for the detection of aging chondrocytes. Alcian Blue staining and 尾 -galactosidase staining of P1 passage cells in each group had no significant difference. The resveratrol group of 2.5 渭 mol/L in P3P5 passage was significantly more deeply stained than other groups. The number of senescent chondrocytes was significantly decreased. The expression of Aggrecan and SIRT1 genes in the group of p53p21 and P16 expression of 2.5 渭 mol/L Resv was significantly up-regulated by RT-PCR, and the expression of P53P21 and P16 was down-regulated by RT-PCR. In conclusion, resveratrol at low concentration (2.5 渭 mol/L) may inhibit the dedifferentiation of chondrocytes by upregulating SIRT1 and down-regulating the expression of P53P21 / P16 gene.
【作者单位】：南昌大学第二附属医院骨科;南昌大学研究生院医学部;九江市武宁县人民医院;九江学院九江市转化医学重点实验室;
【基金】：国家自然科学基金(81160226)
【分类号】：R684

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