APN基因表达的调控在OP骨代谢疾病中的作用机制研究

Genetic gene is closely related with osteoporosis(OP) which seriously hazard human health; our results of the previous project of National Nature Science fundation(NO.30860117) show that the polymorphisms of adiponectin (APN,Acrp30,AdipoQ,apM1,GBP28) gene is closely related to OP in the elderly pepole of Zhuang ethnic groups in Guangxi province. However it is unclear that what role the regulation of APN gene expression plays in the origin and development of OP. Up to now, the published documents about the mechanisms are rarely. Based on the results of our previous research, we will detect the wide-band ultrasonic attenuation(BUA) of calcaneal ; and genotypes of the APN gene and plasma APN protein levels will be measured by TaqMan-MGB probe technology and ELISA respectively; Lipid levels will be measured by automatic biochemical analyzer. Fat mass, muscle mass will be measured by body composition analyzer. Bone marrow fat will be measured by MRI.APN mRNA and APN protein in mononuclear cells from peripheral blood of samples with different genotypes and APN protein in the periosteum of the samples with osteoporotic fracture will be detected with quantitative real-time PCR and western blot respectively. And APN protein levels in osteoblasts and osteoclasts of periosteal will be detected by histological techniques and the number of osteoclasts in periosteal will be counted also. We will compare the differences of different genotype samples. The results will be used to analysis the effect of APN gene polymorphism on its mRNA and protein expression, and further to reveal the molecular mechanisms of regulation of APN gene expression in the origin and development of OP. Illuminating the molecular mechanisms will makes a significant contribution to the prevention and treatment of OP.

遗传基因与严重危害人类健康的骨质疏松症（OP）关联密切，在上个国家自然科学基金中我们发现脂联素（APN）基因多态性与OP密切相关，但APN基因表达的调控在OP发生发展中的作用机制尚不清楚，截至目前鲜见该方面的文献报道。在已有的前期基础上，本课题采用TaqMan-MGB 探针技术对APN基因进行分型，用ELISA 测定血浆脂联素水平，测定血脂水平、脂肪量、肌肉量、内脏脂肪量及骨髓脂肪量，用RT-PCR和western blot技术分别检测不同基因型样本外周血单个核细胞中APN mRNA和蛋白的表达水平及骨质疏松性骨折样本骨膜中脂联素水平，用组织学技术检测骨膜破骨细胞数量及成骨细胞和破骨细胞的脂联素水平，比较不同基因型样本间差异。多层面分析APN基因多态性对其mRNA和蛋白表达的影响，探讨APN基因表达的调控功能，揭示APN 基因表达的调控在OP发生发展中的分子作用机制，为防治OP提供参考。

遗传基因与严重危害人类健康的骨质疏松症（OP）关联密切，在上个国家自然科学基金中我们发现脂联素（APN）基因多态性与OP密切相关，但APN基因表达的调控在OP发生发展中的作用机制尚不清楚，截至目前鲜见该方面的文献报道。在已有的前期基础上，随机整群抽取广西40-90岁无血缘关系772个壮族人为研究对象。应用超声骨密度测量仪检测跟骨骨密度(BMD)，计算T值，确定骨量正常或减少。测定血脂水平，ELISA测定血清APN水平，采用iMLDR技术对APN基因进行分型，体成分分析仪测定脂肪量、肌肉量、内脏脂肪量，用RT-PCR和western blot技术分别检测不同基因型样本外周血单个核细胞中APN mRNA和蛋白的表达水平及骨质疏松性骨折样本骨膜中APN水平，用组织学技术检测骨膜破骨细胞数量及成骨细胞和破骨细胞的APN水平，比较不同基因型样本间差异。结果：（1）骨质疏松检出率男性27.7%、女性64.2%，骨量减少检出率男性29.3%、女性14.4%。BMD与APN、高密度脂蛋白弱负相关，其与甘油三脂、极低密度脂蛋白正相关。骨量减少组的脂联素浓度高于对照组的，其低密度脂蛋白（LDL-C）也是高于对照组的。LDL-C和APN基因第3外显子rs1063539多态性与壮族人群骨量减少有一定关联。rs1063539 CC型对骨密度正常具有一定的保护作用,CG型和GG型是骨量减少的危险因素。LDC-C水平升高发生骨量减少的危险增加。（2）不同性别中老年RSMI均与BMD显著正相关。壮族男性骨骼肌减少与骨质疏松存在明显的相关性。增加骨骼肌质量对中老年人群骨质疏松的防治有重要实践意义。（3）高龄、女性、吸烟、有骨折且伴有高血压或糖尿病的、抗骨质疏松药物治疗依从性差的骨质疏松性骨折患者其骨折再发率较高。（4）不同基因型样本外周血单个核细胞中APN mRNA和蛋白的表达水平及骨质疏松性骨折样本骨膜中脂联素水平存在差异，骨膜中破骨细胞数量及成骨细胞和破骨细胞的脂联素水平存在差异。本研究多层面探讨APN基因表达的调控功能，揭示APN基因表达的调控在OP发生发展中的分子作用机制，为防治OP提供参考。