重组人骨形态发生蛋白2缓释体影响细胞外基质金属蛋白酶诱导因子的表达

Effect of recombinant human bone morphogenetic protein-2 slow-release delivery system on extracellular matrix metalloproteinases expression

背景:目前,重组人骨形态发生蛋白2是否可以抑制炎性细胞因子的产生及活性,从而抑制溶骨反应的分子生物学机制仍不清楚。目的:观察重组人骨形态发生蛋白2缓释体在大鼠皮下气囊中的分子生物学反应,探讨人骨形态发生蛋白2抑制骨溶解生物活性的可能机制。方法:于鼠背部皮下注射过滤后的空气3mL,每2d1次,共6次。1周后,20只SD大鼠按注射成分的不同随机分为2组,每组10只。分别于囊腔内注射人骨形态发生蛋白2缓释体悬液(实验组)和生理盐水(对照组)。分别于1,2周后取囊腔组织进行光镜下观察,测定血清碱性磷酸酶水平;real-time PCR法检测细胞外基质金属蛋白酶诱导因子(EMMPRIN)的mRNA水平;Western blotting法测定EMMPRIN的蛋白表达。结果与结论:光镜下实验组与对照组囊腔组织均未见明显组织细胞反应。实验组第1周血清碱性磷酸酶水平与实验组第2周及对照组相比,差异具有显著性意义(P<0.05),实验组第2周较对照组略低,但差异无显著性意义(P>0.05)。实验组EMMPRIN的mRNA及蛋白水平明显低于对照组,而且第2周小于第1周。提示重组人骨形态发生蛋白2缓释体抑制了溶骨性细胞因子的产生及活性,从而抑制溶骨反应。

BACKGROUND：It has been still unclear that whether recombinant human bone morphogenetic protein-2（rhBMP-2） may inhibit production and activity of inflammatory factors,thereby inhibits molecular biomechanism of osteolytic reaction.OBJECTIVE：To investigate the subcutaneous molecular biological reaction of rhBMP-2 slow-release delivery system in the rat air pouch model,and to analyze the possible mechanism of the inhibition of osteolysis indueced by rhBMP-2.METHODS：Filtration air（3 mL） was subcutaneously injected into back of rats,once every two days,6 times in total.One week later,20 rats were randomly divided into 2 groups,with 10 rats per group.rhBMP-2 low-release formulation and saline were individually injected into air pouch of experimental and control groups.Air pouch tissue was obtained one or two weeks later to measure serum alkaline phosphatase level under optic microscope.Real-time PCR was used to detect extracellular matrix metalloproteinases mRNA level,and Western blotting was used to detect its protein expression.RESULTS AND CONCLUSION：No tissular and cellular reactions were found in both experimental and control groups.Serum alkaline phosphatase level in the experimental group in 1 week was significantly greater than in experimental group in 2 weeks and control group（P 0.05）,while the level in the experimental group in 2 weeks was less than in the control group,but there was no significant difference（P 0.05）.Extracellular matrix metalloproteinases mRNA and protein expressions in the experimental group were significantly less than in the control group,while the level in 1 week was less than in 2 weeks.The results suggested that rhBMP-2 slow-release delivery system inhibited the production and activities of osteolytic cytokines,so as to inhibit osteolytic reaction.