摘要
目的 探讨胡黄连苷Ⅱ调节受体相互作用蛋白激酶(RIP)1/RIP3/混合系列蛋白激酶样结构域(MLKL)信号对地塞米松(DEX)诱导成骨细胞凋亡的影响.方法 体外培养hFOB 1.19细胞,随机分为对照组、DEX组、DEX+胡黄连苷Ⅱ组、DEX+RIP1过表达组、DEX+空载质粒组、DEX+胡黄连苷Ⅱ+RIP1过表达组,分组处理后检测各组细胞活力、凋亡率、活性氧(ROS)相对水平、乳酸脱氢酶(LDH)、白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)及IL-6释放量、BCL-2及BAX相对表达、RIP1/RIP3/MLKL信号蛋白表达.结果 与对照组相比,DEX组细胞活力、BCL-2相对表达降低(P＜0.05),凋亡率、ROS相对水平、LDH及IL-1β、TNF-α、IL-6释放量、细胞BAX及RIP1、RIP3、MLKL蛋白相对表达升高(P＜0.05).与DEX组相比,DEX+胡黄连苷Ⅱ组细胞活力、BCL-2相对表达升高(P＜0.05),凋亡率、ROS相对水平、LDH及IL-1β、TNF-α、IL-6释放量、细胞BAX及RIP1、RIP3、MLKL蛋白相对表达降低(P＜0.05);DEX+RIP1过表达组细胞活力、BCL-2相对表达降低(P＜0.05),凋亡率、ROS相对水平、LDH及IL-1β、TNF-α、IL-6释放量、细胞BAX及RIP1、RIP3、MLKL蛋白相对表达升高(P＜0.05);DEX+空载质粒组细胞各指标差异无统计学意义(P＞0.05);过表达RIP1可减弱胡黄连苷Ⅱ对DEX诱导的hFOB 1.19细胞的作用.结论 胡黄连苷Ⅱ可通过下调RIP1/RIP3/MLKL通路而减轻炎症,从而抑制DEX诱导的成骨细胞凋亡.
Abstract
Objective To investigate the effect of picroside Ⅱ on dexamethasone(DEX)-induced osteoblast apoptosis by regulating the receptor-interacting protein(RIP)l/RIP3/mixed lineage kinase domain-like(MLKL)signal.Methods HFOB 1.19 cells were cultured in vitro and randomly grouped into control group,DEX group,DEX+picroside Ⅱ group,DEX+RIP 1 overexpression group,DEX+empty plasmid group,and DEX+picroside Ⅱ+RIP1 overexpression group.After grouping treatment,cell viability rate,apoptosis rate,the relative level of reactive oxygen species(ROS),the releases of lactate dehydrogenase(LDH),interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-6,the relative expressions of BCL-2 and BAX,the expressions of RIP1/RIP3/MLKL signal proteins were detected in cells of each group.Results Compared to those in the control group,the cell viability and the relative expression of BCL-2 in DEX group decreased(P＜0.05),the apoptosis rate,ROS relative level,LDH and IL-1β,TNF-α,IL-6 releases,and the relative expressions of BAX and RIP1,RIP3,MLKL proteins increased(P＜0.05).Compared to those in DEX group,the cell viability and the relative expression of BCL-2 in DEX+picroside Ⅱ group increased(P＜0.05),the apoptosis rate,ROS relative level,LDH and IL-1β,TNF-α,IL-6 releases,and the relative expressions of BAX and RIP1,RIP3,MLKL proteins decreased(P＜0.05).The cell viability and the relative expression of BCL-2 in the DEX+RIP1 overexpression group decreased(P＜0.05),the apoptosis rate,ROS relative level,LDH and IL-1β,TNF-α,IL-6 releases,and the relative expressions of BAX and RIP1,RIP3,MLKL proteins increased(P＜0.05).There was no obvious difference in cell indexes in DEX+empty plasmid group(P＞0.05).Overexpression of RIP1 weakened the effect of picroside Ⅱ on DEX induced hFOB 1.19 cells.Conclusion Picroside Ⅱ reduces inflammation by down-regulating RIP1/RIP3/MLKL pathway,thus inhibiting the apoptosis of osteoblasts induced by DEX.
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