摘要
目的观察成骨细胞特异性识别多肽对人颅骨成骨细胞(HCO)增殖和矿化作用的影响。方法体外常规培养人颅骨成骨细胞,分为5个实验组(10^(-4)、10^(-5)、10^(-6)、10^(-7)、10^(-8)mol/L)以及空白对照组,MTT法检测细胞的增殖,紫外分光光度法检测细胞碱性磷酸酶(ALP)的表达,茜素红染色及半定量分析检测细胞矿化程度,RT-QPCR法检测细胞骨钙素(OCN)mRNA的表达。结果此浓度范围内的特异性多肽对成骨细胞增殖有促进作用(P<0.05),最大效应浓度是10^(-5)mol/L;能增加ALP活性(P<0.05),最大效应浓度是10^(-4)mol/L。通过茜素红染色观察和半定量分析,14、21 d实验组钙盐沉积量高于对照组,半定量分析结果显示浓度10^(-5)mol/L时,吸光度(OD)值最高,但差异无统计学意义;RT-QPCR法显示14、21 d实验组OCN mRNA表达量均高于对照组(P<0.05)。结论 10^(-8)~10^(-4)mol/L浓度范围的成骨细胞特异性识别多肽能够促进人颅骨成骨细胞的增殖和矿化,且在10^(-5)mol/L浓度促进成骨细胞增殖及矿化相关蛋白表达最为显著。
Objective To investigate the effect of the peptide that specifically bind to osteoblasts on proliferation and mineralization of human ealvarial osteoblasts in vitro. Methods Human calvarial osteoblasts were routting cultured which were divided into 6 groups including experimental groups containing the peptide at various concentrations（ 10-4,10-5, 10-6,10-7 ,10-8 tooL/L）and a control group. The proliferative activity was examined by MTF, alkaline phosphatase activity was examined by ultraviolet spectrophotometry, the degree of cell mineralization was examined by alizarin red staining and semi-quantitative analysis, and RT-QPCR test was used to detect RNA level of OCN. Results Compared with the control group, the proliferation and alkaline phosphatase activity in different peptide groups were increased（P 〈 0. 05）, and the maximum response was obtained with 10-5 moL/L and 10 4 mol/L. Alizarin red staining cells cultured for 14 and 21 d, the deposition of the calcium in experimental group （10-5 mol/L） were higher than the control group, but there was no statistically significant difference. The results of RT-QPCR test showed that mRNA level of OCN in experimental groups was much higher than the control group （P 〈 0.05 ）. Conclusion The peptide that specifically bind to osteoblasts in the concentrations of 10-8 mol/L to 10 -4 mol/L could promote the proliferation and mineralization of human calvarial osteoblasts in vitro, and the best concentration is 10-5 mol/L.
出处
《安徽医科大学学报》
CAS
北大核心
2016年第3期337-340,共4页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81170993)
关键词
成骨细胞特异性识别多肽
人颅骨成骨细胞
增殖
矿化
the peptide that specifically bind to osteoblasts
human calvarial osteoblasts
proliferation
mineralization
