红景天苷通过JAK2/STAT3通路抑制退变髓核细胞的炎症反应和凋亡

Salidroside inhibits inflammatory response and apoptosis of degenerated nucleus pulposus cells through JAK2/STAT3 signaling pathway

目的观察红景天苷对退变髓核细胞凋亡和炎症反应的影响,并探究其与JAK2/STAT3信号通路的关系。方法通过氧糖剥夺(oxygen glucose deprivation,OGD)培养建立退变髓核细胞模型,退变细胞分别予浓度为10μg/mL、30μg/mL和50μg/mL的红景天苷培养液。后续实验选择30μg/mL浓度进行,分为Control组、OGD组、红景天苷组、AG490(JAK2/STAT3信号通路抑制剂)组和红景天苷联合AG490干预组。RT-PCR法检测髓核细胞蛋白聚糖(Aggrecan)和Ⅱ型胶原(Col2a1)表达水平;Western blot检测红景天苷对髓核细胞p-JAK2和p-STAT3蛋白水平的影响;ELISA检测上清液中炎症相关因子TNF-α、IL-1β和IL-6的表达水平,流式细胞术检测髓核细胞的凋亡。结果与Control组相比,OGD组Aggrecan和Col2a1 mRNA表达明显降低,炎症相关因子TNF-α、IL-1β和IL-6水平升高,p-JAK2和p-STAT3的蛋白水平明显上调。与OGD组相比,红景天苷组显著抑制细胞凋亡以及炎症相关因子TNF-α、IL-1β和IL-6的释放,同时,我们发现,AG490组和红景天苷联合AG490干预组上述指标均明显降低,其中红景天苷联合AG490干预组各检测指标较AG490组略低。红景天苷联合AG490干预明显改善髓核细胞的凋亡和炎症反应,并抑制JAK2和STAT3的磷酸化水平。结论红景天苷可能通过抑制JAK2/STAT3信号通路的活化而减轻退变髓核细胞炎症因子的产生和细胞凋亡。

Objective To observe the effect of salidroside on apoptosis and inflammatory response of degenerated nucleus pulposus cells and to explore its relationship with JAK2/STAT3 signaling pathway. Methods The degenerated nucleus pulposus cell model was established by oxidative glucose deprivation（ OGD） culture. The degeneration cells were cultured in addition of 10μg/mL,30 μg/mL,or 50 μg/mL of salidroside,respectively. Cells in 30 μg/mL of salidroside were used in the follow-up experiments. They were divided into control group,OGD group,salidroside group,AG490（ inhibitor of JAK2/STAT3 signaling pathway） group,and salidroside combined with AG490 intervention group. The mRNA expression of aggrecan and Col2 a1 in nucleus pulposus cells was detected by RT-PCR. Western blotting was used to detect the effects of salidroside on p-JAK2 and pSTAT3 protein levels in nucleus pulposus cells. The levels of TNF-α,IL-1β,and IL-6 in the supernatant were detected by ELISA.The apoptosis of nucleus pulposus cells was detected by flow cytometry. Results Compared with control group,the mRNA expression of aggrecan and Col2 a1 in OGD group decreased significantly,but the levels of TNF-α,IL-1β,and IL-6 increased,and the protein levels of p-JAK2 and p-sTAT3 were significantly up-regulated. Compared with OGD group,salidroside significantly inhibited release of apoptosis and inflammation-related factors TNF-α,IL-1β and IL-6. Meanwhile,we found that the above indicators in AG490 group and salidroside combined with AG490 intervention group decreased significantly,and those in salidrosidecombined with AG490 intervention group were slightly lower than those in AG490 group. Salidroside combined with AG490 pretreatment significantly relieved the apoptosis and inflammation response of nucleus pulposus cells and inhibited the phosphorylation of JAK2 and STAT3. Conclusion Salidroside reduces the apoptosis and inflammation response of degenerative nucleus pulposus cells by inhibiting the activation of JAK2/STAT3 signaling pathway.