瞬时感受器电位离子通道香草素受体亚家族Ⅳ在帕金森病细胞模型中介导PC12细胞炎症反应的机制

Mechanism of transient receptor potential vanillin receptor subfamilyⅣmediating inflammation of PC12 cells in Parkinson's disease cell model

目的探讨瞬时感受器电位离子通道香草素受体亚家族Ⅳ(TRPV4)在帕金森病(PD)细胞模型中介导PC12细胞炎症反应的机制。方法2023年6―8月,用1-甲基-4-苯基吡啶离子(MPP^(+))建立PD细胞模型,用TRPV4特异性抑制剂HC067047抑制TRPV4。将培养的PC12细胞采用随机数字表法分为四组:对照组、HC067047组、MPP^(+)组、HC067047+MPP^(+)组。细胞毒性检测试剂盒(CCK-8)检测各组细胞的增殖活力。蛋白印迹法和酶联免疫吸附测定(ELISA)检测各组细胞TRPV4和炎症因子白细胞介素-18(IL-18)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的水平变化。结果与对照组1.00±0.08相比,MPP^(+)组TRPV4的表达量2.14±0.20明显升高(P<0.001)。与对照组1.00±0.01相比,MPP^(+)组的PC12细胞活力0.65±0.08明显降低(P<0.01),而HC067047能明显回复MPP^(+)引起的细胞活力0.83±0.07降低(P<0.01)。与对照组相比,MPP^(+)组的IL-181.96±0.27和IL-61.92±0.18、IL-1β(874.61±108.09)ng/L和TNF-α(791.28±106.88)ng/L明显升高(P<0.001),HC067047能明显抑制MPP^(+)引起的IL-181.45±0.11和IL-61.58±0.22、IL-1β(626.28±84.53)ng/L和TNF-α(592.94±86.9)4 ng/L明显升高(P<0.01或P<0.05)。结论TRPV4参与了MPP^(+)诱导的PD细胞模型的炎症反应,抑制TRPV4有抗炎作用。

Objective To investigate the mechanism of transient receptor potential vanillin receptor subfamilyⅣ(TRPV4)mediating inflammation of PC12 cells in Parkinson's disease(PD)cell model.Methods TRPV4 specific inhibitor HC067047 was used to inhibit TRPV4 in 1-methyl-4-phenylpyridinium(MPP^(+))-induced cell model of Parkinson's disease from June to August 2023.PC12 cells were randomly divided into four groups:control group,HC067047 group,MPP^(+)group and HC067047+MPP^(+)group.Cell Counting Kit-8(CCK-8)assays were used to detect the cell viability of each group.The expressions of TRPV4 and the levels of inflammatory factors,such as interleukin(IL)-18,IL-6,IL-1βand tumor necrosis factor-α(TNF-α)were detected by western blot and ELISA.Results Compared with the control group 1.00±0.08,the expression of TRPV4 in MPP^(+)group 2.14±0.20 was increased(P<0.001).Compared with the control group 1.00±0.01,the cell viability of MPP^(+)group 0.65±0.08 was decreased obviously(P<0.01),while HC067047(0.83±0.07)could restore the decrease of cell viability caused by MPP^(+)(P<0.01).Additionally,the levels of IL-181.96±0.27,IL-61.92±0.18,IL-1β(874.61±108.09)ng/L and TNF-α(791.28±106.88)ng/L in MPP^(+)group were increased significantly(P<0.001),while HC067047 could obviously inhibit the high levels of IL-181.45±0.11,IL-61.58±0.22,IL-1β(626.28±84.53)ng/L and TNF-α(592.94±86.94)ng/L induced by MPP^(+)(P<0.01 or P<0.05).Conclusion TRPV4 participates MPP^(+)-induced inflammatory response in PD cell model,inhibiting the expression of TRPV4 has anti-inflammatory effect.