论文部分内容阅读
目的 观察磷酸三钙(tricalciumphosphate,TCP)颗粒与锌(zinc,Zn)掺杂TCP(ZnTCP)颗粒对骨细胞MLO-Y4活性和功能影响及Zn对TCP颗粒诱导骨细胞损伤的作用.方法 骨细胞MLO-Y4分别与TCP颗粒(0.1 mg/ml)和ZnTCP颗粒(0.1 mg/ml)共培养,应用实时荧光定量PCR和Western blot等方法 检测Zn干预后骨细胞硬化蛋白SOST(sclerostin)和牙本质基质蛋白1(dentin matrix protein 1,DMP-1)mRNA水平及蛋白表达的变化,研究Zn对TCP磨损颗粒诱导骨细胞功能损伤的影响;通过Calcein-AM标记、MTT和流式细胞术定量分析Zn对TCP颗粒诱导骨细胞活性和凋亡的影响;利用Western blot方法 检测Zn干预后骨细胞Akt、phospho-Akt(p-AktSer473)和phospho-Akt(p-AktThr308)等蛋白的表达变化,探讨Akt信号分子在Zn对TCP颗粒诱导骨细胞损伤保护中的作用,阐明其分子机理.结果 Zn可明显抑制TCP颗粒诱导的骨细胞功能损伤,阻止骨细胞凋亡;同时Zn干预能显著减弱TCP颗粒诱导的骨细胞Akt蛋白失活,表现为上调p-Aktser473和p-AktThr308等蛋白表达(P<0.05).结论 Zn可阻止TCP颗粒诱导的骨细胞损伤,可能与抑制TCP颗粒诱导骨细胞Akt蛋白的失活密切相关.“,”Objective To observe the effect of tricalciumphosphate (TCP) particles and zinc-containing TCP (ZnTCP) particles on osteocytes MLO-Y4, and to examine the effect of Zn on osteocytes which were injured by TCP paritcles. Methods MLO-Y4 cells and TCP particles (0.1 mg/ml)/ ZnTCP particles (0.1 mg/ml) were co-incubated. The mRNA and protein expressions of SOST and DMP-1 of osteocytes were detected by real-time quantitative PCR and Western blot. Osteocytes injuries were assessed using calcein-AM labeling, MTT assay and the flow cytometry, respectively. The expressions of Akt, phospho-Akt (p-AktSer473) and phospho-Akt (p-AktThr308) of osteocytes were showed by Western blot. Results Zn significantly inhibited TCP particles-induced dysfunction of osteocytes MLO-Y4, and also significantly prevented osteocytes apoptosis (P<0.05). Furthermore, Zn greatly increased Akt activation which was inhibited by TCP particles, and up-regulated the levels of p-AktSer473 and p-AktThr308 (P<0.05).Conclusion Zn may prevent TCP particles-induced osteocytes injury, which is mediated by increasing activation of Akt profein in osteocytes MLO-Y4.