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论文编号:
贵阳护理职业学院
2012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
2012 年 6 月
目录
中文摘要 IV
英文摘要 V
前言 1
1. BMP蛋白 2
BMP结构和功能 2
BMP诱导成骨机制 2
2. Noggin基因的概况 3
Noggin基因的结构 3
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 ic 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***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(P<),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2×10-9 mol/L,℅;noggin为8×10-9mol/L,℅;IGF-I为1×10-8mol/L,℅。
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 ic protein 2 (BMP2) and insulin-like growth factor I (IGF-I) on the proliferation rates and differentiation of osteoblasts.
Methods: The human a 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 different time points (12h, 24h, 48h, 72h) after optimal dosage of noggin BMP2, IGF-I were added into the culture 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 pared with that of CON group(