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.论文编号： 贵阳护理职业学院2014届高职高专学生毕业论文Noggin、BMP2和IGF-I对成骨细胞生长的调节作用系 别： 专 业： 指导老师： 学 生： 中国贵州贵阳 2013 年 6 月目 录中文摘要IV英文摘要V 前 言11. BMP蛋白21.1 BMP结构和功能21.2 BMP诱导成骨机制22. Noggin基因的概况32.1 Noggin基因的结构32.1.1 Noggin基因结构特征4 结 果25致 谢51参 考 文 献52中文摘要目的：研究noggin、骨形态发生蛋白2（bone morphogenetic protein 2，BMP2）、胰岛素样生长因子I（insulin like growth factor I，IGF-I）对成骨细胞增殖和分化的调节作用。方法：1、将人成骨肉瘤细胞MG63于RPMI 1640培养基中培养，添加不同浓度的noggin、BMP2、IGF-I蛋白进行药物干预，用四甲基偶氮唑盐（methylthiazolyl-tetrazolium，MTT）法研究细胞的增殖率。2、选用最适浓度的noggin、BMP2、IGF-I蛋白作用成骨细胞，运用ELISA方法和RT-PCR技术分别检测成骨细胞分化标志物骨碱性磷酸酶（bone alkaline phosphatase, BALP）和骨钙蛋白（osteocalcin, OC）的蛋白含量和mRNA的表达情况。结果：1、各药物干预组OD值与对照组相比有统计学差异（P0.05），对成骨细胞增殖的作用效应呈剂量和时间依赖性，其作用的最佳时间均在72h。各药物组内之间比较，确定了最佳作用浓度，BMP2为210-9 mol/L，对MG63细胞的增殖率达179.59；noggin为810-9mol/L，对其增殖的抑制率达到15.51；IGF-I为110-8mol/L，其增殖率达189.80。结论：BMP2、IGF-I对MG63细胞增殖有促进作用，而noggin则抑制MG63细胞的增殖。Noggin、BMP2和IGF-I促进BALP的分泌。Noggin对OC蛋白的分泌有抑制作用，而BMP2、IGF-I则促进OC蛋白的分泌。关键词：Noggin蛋白，BMP2蛋白，IGF-I蛋白，成骨细胞，增值，分化英文摘要Object: The aims is to study the effects of noggin, bone morphogenetic protein 2 (BMP2) and insulin-like growth factor I (IGF-I) on the proliferation rates and differentiation of osteoblasts.Methods: The human osteosarcoma cell line MG63 were chosen using the RPMI1640 medium with 10 fetal bovine serum (FBS). Then, the cultured MG63 cells were exposed to BMP2, noggin and IGF-I at different concentrations with zero concentration as control group (CON). Ability of the proliferation of MG63 cells was detected by methylthiazolyltetrazolium method (MTT). the concentration of BALP and OC of osteoblasts were determined by ELISA method.at different time points (12h, 24h, 48h, 72h) after optimal dosage of noggin BMP2, IGF-I were added into the culture medium.And the expression level of BALP and OC were investigated by reverse transcription PCR at the same time.Results: 1. The proliferation ability of MG63 cells treated with BMP2, noggin and IGF-I groups were significantly different compared with that of CON group(P0.05). The effects of three factors on MG63 cell was dose-dependently and time-dependently after cultured for 72h. The most effective growth of MG63 cells showed at 210-9 mol/L of BMP2 with the proliferation rate of 179.59 and 110-8mol/L of IGF-1 with the rate of 189.80, while 810-9mol/L noggin with the inhibitory proliferation rate of 15.51, respectively. 2. The concentration of BALP treated with BMP2, noggin and IGF-I groups were significantly different compared with that of CON group (P0.05) for 48h, especially, the noggin group was extremely significant (P0.01), and the BALP mRNA expression were also significantly different (P0.05). 3. The concentration of OC treated with noggin groups were significantly different compared with that of CON group (P0.01) for 24h, but the OC mRNA expression were not significantly different (P0.05). The concentration of OC of BMP2 group were significantly different compared with that of CON group significantly different compared with that of CON group (P0.01) , and the OC mRNA expression were also significantly different (P0.05). The concentration of OC of IGF-I group were significantly different compared with that of CON group significantly different compared with that of CON group (P0.01), and the OC mRNA expression were also significantly different (P0