摘要
目的探讨钛合金微粒(Ti-6Al-4V)对smad4、smad5、smad8影响,了解微粒对促骨形成信号通路影响。方法根据Ti-6Al-4V微粒浓度高低,将成骨细胞分为正常对照组(0μg/mL)、Ti-6Al-4V组(5、10、15μg/mL),采用RT-PCR和蛋白免疫印迹法(Western Blot)检测培养72h成骨细胞smad4、smad5、smad8 mRNA和蛋白表达。结果培养72h后,与对照组比较,smad 4、smad5、smad8 mRNA水平下降,差异有统计学意义(P<0.05),smad4 mRNA水平随Ti-6Al-4V微粒浓度增加呈抑制作用,差异有统计学意义(P<0.05),smad4、smad5、smad8蛋白水平分别与其mRNA表达变化趋势一致。结论 Ti-6Al-4V微粒下调受体调节蛋白smad5、smad8和公用型蛋白Smad4表达,是Ti-6Al-4V微粒抑制核转录因子runx2表达关联因素之一。
Objective Ti-6A1-4V alloy particles inhibited the expression of Runt-related transcription factor 2 （runx2） in osteoblast, but it was unclear whether it was a result of upstream of runx2 gene. This study aimed to investi- gate the effects of Ti-6A1-4V on Smad4, Smad5 and SmadS, and to explore the effect of signal pathway of bone formation. Methods Osteoblasts were cultured in media containing different concentrations of Ti-6A]-4V particles. The control group （0 p.g/mL） was cultured in medium without Ti-6A1-4V particles, while osteoblast Ti-6A1-4V group was cultured in medium containing Ti-6AI-4V particles at 5 p^g/mL, l0 p.g/mL and 15 p^g/mL, respectively. $madsprotein including Smad4, Smad5 and Smad8 mRNA levels and their protein expressions were detected by reverse transcription polymerase chain reaction and western-blotting, respectively during 72 h. Results After 72 h incubation, compared to the control group, the $mad4, Smad5 and Smad8 mRNA levels in the Ti-6A1-4V group significantly decreased （ P 〈 0. 05 ）. Smad4 mRNA levels had the obvious inhibiting effect with the increase of Ti-6A1-4V particles concentration （P 〈 0. 05 ）. The protein levels of Smad4, 5mad5 and Smad8 were also down-regulated by Ti-6AI-4V particles, which was consistent with the change in their mRNA levels. Conclusion The common-mediator Smad4, the receptor-regulated Smad5 and Smad8 mRNA and protein levels were significantly down-regulated by Ti-6A1-4V particles, suggesting that the suppression of Runx2 expression was probably related to Smad4, Smad5 and Smad8 in osteoblast-indueed Ti-6A1-4V particles.
出处
《中华骨质疏松和骨矿盐疾病杂志》
2015年第2期148-151,共4页
Chinese Journal Of Osteoporosis And Bone Mineral Research
基金
江苏省临床医学科技专项(BL2012002)
南京军区医学科技创新项目(11z028)
南京市科技发展项目(2012sc311020)
