白藜芦醇通过上调SIRT1促进退变髓核细胞胞外基质合成

Resveratrol stimulates extracellular matrix synthesis in degenerative nucleus pulposus cells via upregulation of SIRT1

目的:研究白藜芦醇(RES)对人退变椎间盘髓核细胞(DNPCs)合成细胞外基质(ECM)的影响。方法:对人退变椎间盘髓核组织进行分离、单层培养细胞鉴定、藻酸盐培养。取原代藻酸盐培养细胞分别用0、12.5、25、50、100、200μmol/L的RES分别刺激DNPCs 12、24、48 h,Western blot法检测沉默交配型信息调节因子2同源蛋白1(silent mating typeinformation regulation 2 homolog 1,SIRT1)、Ⅱ型胶原(Colla2α1)、蛋白聚糖聚合物(aggrecan)蛋白表达,real-time荧光定量PCR检测SIRT1 mRNA表达水平。用SIRT1-siRNA转染后再与100μmol/L RES共培养24 h,观察Colla2α1、aggrecan的蛋白表达。结果:RES上调SIRT1 mRNA和蛋白水平的表达,促进DNPCs合成ECM的表达,与对照组相比具有统计意义(P<0.05)。siRNA靶向沉默SIRT1表达后,再加入RES刺激,观察到Colla2α1与aggrecan的蛋白表达与对照组相比显著下降(P<0.05)。结论:RES可刺激DNPCs ECM的合成,且呈现一定的量效关系,这种调节作用与SIRT1的活性有关。

AIM： To study the impact of resveratrol （RES） on the synthesis of extracellular＇matdx （ECM） in de- generative nucleus pulpceus cells （ DNPCs ）. METHODS： Human degenerative nucleus pulposus tissues were isolated, identified through monolayer culture, and cultivat- ed in alginate. Primary alginate-cultured DNPCs were irritat- ed by 0, 12.5, 2.5, 50, 100 and 200 μmol/L RES for 12, 24 and 48 h, respectively. The protein expressions of silent mating type information regulation 2 homolog I （SlRT]）, Colla2al and aggrecan were examined by Western blotting, and the expression level of SlRT1 mRNA was measured through real-time fluorescence quantitative PCR. The cells were transfected by SlRTI-siRNA and cultured in 100 μmol/L RES for 24 h. Then the protein expressions of Colla2a1 and aggrecan were observed. RESULTS.. RES up-regulated the expressions of SIRT1 at mRNA and protein levels, and promoted the expression of DNPCs-synthesized ECM, which were significantly different from the control group （P〈0.05）. After the expression of SlRT1 was silenced by siRNA, RES was ＇added for irritation. The protein expres- sions of Colla2a1 and aggrecan were significantly reduced in comparison with the control group （ P 〈 0.05）. CONCLU- SION： RES can stimulate the synthesis of ECM in DNPCs, which exhibits a dose-effect relationship. This regulating effect is related to the activity of SIRT1.