阻断Cn/NFAT通路对钛颗粒诱导成骨细胞分泌IL-6和PGE_2的影响

Blockade of calcium phosphatase(Cn) / activated T nuclear factor(NFAT) pathway by 11R-VIVIT peptide inhibits IL-6 and PGE_2 expression in wear particles induced osteoblast cells

目的:观察阻断钙调磷酸酶(Cn)/活化T细胞核因子(NFAT)通路对钛颗粒诱导成骨细胞分泌IL-6和PGE2的影响。方法:取新生大鼠头颅骨,经消化法获取成骨细胞,根据培养条件,分为钛颗粒组、钛颗粒/VIVIT肽组、VIVIT肽组和对照组。细胞培养96 h后行Western blot检测NFATc1蛋白表达,于6、24、96 h离心后取细胞培养上清,用ELISA法检测各组IL-6和PGE_2的含量。结果:钛颗粒组NFATc1表达高于对照组(P<0.01),钛颗粒/VIVIT肽组NFATc1表达显著低于钛颗粒组(P<0.01),钛颗粒组培养上清IL-6和PGE_2含量各时间点均显著高于对照组(P<0.05),钛颗粒/VIVIT肽组较钛颗粒组各时间点均显著降低(P<0.05)。结论:11R-VIVIT肽特异性阻断Cn/NFAT通路可显著抑制钛颗粒诱导的成骨细胞IL-6和PGE_2的分泌。

Objective To investigate the effects on IL-6 and PGE2 expression in wear-particles-induced osteoblast cells by blocking calcium phosphatase（Cn） / activated T nuclear factor（NFAT） pathway. Methods Fetal Sprague-Dawley rats were used in this study. Osteoblast were prepared from the calvariae of rats. Osteoblast cells were incubated in four group according to different supplementation：（1） neither Ti particles nor 11R-VIVIT（Control group）,（2） only Ti particles（Ti group）,（3） both Ti particles and 11R-VIVIT（Ti / VIVIT group）, and（4） only 11R-VIVIT（VIVIT group）. Cells were incubated for 96 hours and the expression of NFATc1 protein was detected by western blot. The expression of IL-6 and PGE2 in liquid supernatant of osteoblast were detected at 6,24 and 96 hours by ELISA. Results The expression of NFATc1 in the Ti group was higher than that in the Control group（P 〈0.01）, but in Ti / VIVIT group that was significantly lower than in the titanium particle group（P 〈0.01）. The IL-6 and PGE2 expression in the supernatant of the Ti group were significantly increased than those in the control group（P 〈0.05）. The IL-6 and PGE2 in the Ti / VIVIT group were significantly lower than that in the Ti group（P 〈0.05）. Conclusions 11R-VIVIT peptide specific blockade of Cn / NFAT signaling pathway significantly inhibited IL-6 and PGE2 of osteoblast cells induced by titanium particles.