黄芪多糖通过JAK2/STAT3通路调控人膝骨关节炎软骨细胞增殖与凋亡

Astragalus Polysaccharide Regulates the Proliferation and Apoptosis of Human Knee Osteoarthritis Chondrocytes through JAK2/STAT3 Pathway

目的研究黄芪多糖(APS)对人膝骨关节炎软骨细胞增殖、凋亡及JAK2/STAT3通路的影响。方法无菌分离人膝骨关节炎软骨细胞,设空白对照组,APS低、中、高剂量(25、50、100mg/L)组和塞来昔布(20mg/L)组。药物干预48h后,MTT法检测细胞增殖活力,Annexin V-FITC/PI双染法检测细胞凋亡率,PI染色法检测细胞周期分布,Western blot法检测JAK2、p-JAK2、STAT3、p-STAT3、cyclin D1、p21、cleaved caspase-3、Bcl-2、Bax蛋白表达。结果与空白对照组比较,APS中、高剂量组和塞来昔布组细胞增殖活力升高、凋亡率降低(P<0.01),处于G_(0)/G_(1)期细胞比例降低、S期细胞比例升高(P<0.05);p-JAK2、p-STAT3、cyclin D1、Bcl-2表达上调而p21、cleaved caspase-3、Bax表达下调(P<0.01),p-JAK2/JAK2、p-STAT3/STAT3、Bcl-2/Bax比值升高(P<0.01)。与塞来昔布组比较,APS高剂量组细胞增殖活力升高、凋亡率降低(P<0.01);G_(0)/G_(1)期细胞比例降低、S期细胞比例升高(P<0.01);p-JAK2、p-STAT3、cyclin D1表达上调且p21、cleaved caspase-3、Bax表达下调(P<0.01),p-JAK2/JAK2、p-STAT3/STAT3、Bcl-2/Bax比值升高(P<0.01)。结论APS具有促进人膝骨关节炎软骨细胞增殖并抑制其凋亡的作用,其机制可能与激活JAK2/STAT3通路有关。

Objective To investigate the effects of astragalus polysaccharide(APS)on the proliferation,apoptosis and JAK2/STAT3 pathway of human knee osteoarthritis chondrocytes.Methods The human knee osteoarthritis chondrocytes were aseptically isolated,and the blank control group,APS low,medium,high(25,50,100mg/L)dose groups and celecoxib(20mg/L)group were set up.48h after drug intervention,the cell proliferation activity was detected by MTT method,and the cell apoptosis rate was detected by Annexin V-FITC/PI double staining,and the cell cycle distribution was detected by PI staining.The expression of JAK2,p-JAK2,STAT3,p-STAT3,cyclin D1,p21,cleaved caspase-3,Bcl-2,Bax were detected by Western blot.Results Compared with the blank control group,the cell proliferation activity in APS medium,high dose groups and celecoxib group was decreased while the apoptosis rate was decreased(P<0.01).The proportion of cells in G_(0)/G_(1) phase was significantly decreased while in S phase was increased(P<0.05 or P<0.01).The expression of p-JAK2,p-STAT3,cyclin D1,Bcl-2 were up-regulated while the expression of p21,cleaved caspase-3,Bax were down-regulated(P<0.01).The ratio of p-JAK2/JAK2,p-STAT3/STAT3,Bcl-2/Bax were increased(P<0.01).Compared with the celecoxib group,the cell proliferation activity in APS high dose group was decreased while the apoptosis rate was decreased(P<0.01).The proportion of cells in G_(0)/G_(1) phase was decreased while the proportion in S phase was increased(P<0.01).The expression of p-JAK2,p-STAT3,cyclin D1 were up-regulated and the of p21,cleaved caspase-3,Bax were down-regulated(P<0.01).The ratio of p-JAK2/JAK2,p-STAT3/STAT3,Bcl-2/Bax were increased(P<0.01).Conclusion APS can promote the proliferation of human knee osteoarthritis chondrocytes and inhibit its apoptosis,which may be related to the activation of JAK2/STAT3 pathway.