摘要
目的研究重组人骨形态发生蛋白（rhBMP）2和rhBMP13对软骨细胞蛋白多糖基因表达的调控作用。方法采用1，9二甲基亚甲蓝比色法检测高（625ng／m1）、中（125ng／m1）和低（25ng／m1）浓度组的rhBMP2和rhBMP13对软骨细胞硫化糖胺聚糖生成的影响；采用反转录聚合酶链反应测定3种不同浓度组的rhBMP2和rhBMP13对软骨细胞聚集蛋白聚糖mRNA表达的影响；用HoechstDye法检测不同浓度组的rhBMP2和rhBMP13对软骨细胞的增殖作用。结果与对照相比，不同浓度的rhBMP2和rhBMP13促进软骨聚集蛋白聚糖mRNA的表达差异均有统计学意义（P〈0．05）；在DNA的检测中，与对照相比，不同浓度组的rhBMP2和rhBMP13促进软骨细胞增殖差异均无统计学意义。结论rhBMP2和rhBMP13均能增加软骨细胞聚集蛋白聚糖的表达，rhBMP2比rhBMP13对软骨细胞基因表达有更大的正向调控作用。但rhBMP2和rhBMP13对软骨细胞增殖差异无统计学意义。
Objective By exploring the effects of recombinant human bone morphogenetic protein （rhBMP） 2 and rhBMP13 on chondrocytes proteoglycan production and phenotype expression to establish the theoritical mechanisms for the treatment of disc degeneration with chondrocytes transplantation plus BMPs. Methods The dose-dependent effects of rhBMP2 and rhBMP13 on PG protein synthesis and gene expression were detected under different concentrations （0, 25, 125, and 625 ng/ml）. The suffated-glycosaminoglycan （s-GAG） in the culture media and the pericellular matrix was measured with a 1,9-dimethyl-methylene blue （DMMB） colorimetric assay. Reverse transcriptase polymerase chain reaction （RT-PCR） was performed respectively to quantify the relative abundance of aggrecan mRNA. Cell proliferation was examined by Hoechst Dye assay. Results All rhBMP2 and rhBMP13 in different concentrations could significantly increase s-GAG synthesis and gene expression in chondrocytes （P〈0.05）. And at the same concentration, rhBMP2 was more potent than rhBMP13 on s-GAG synthesis. Hoechst Dye assay showed neither rhBMP2 nor rhBMPI3 had significant effect on cell proliferation. Conclusion rhBMP2 and rhBMP13 are able to upregulate s-GAG syn- thesis, in addition, rhBMP2 is more potent than rhBMP13 on aggrecan gene expression regulation, but rhBMP2 and rhBMP13 do not have significant effect on chondrocyte proliferation.
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2009年第2期76-78,共3页
Chinese Journal of Rheumatology
基金
国家自然科学基金（30400163）
