血清中骨质疏松相关蛋白分子标志物的筛选研究

Screening of osteoporosis-related protein markers in serum

目的通过蛋白芯片筛选血清中反映早期绝经后骨质疏松症发生、发展的标志性分子。方法 3月龄雌性SD大鼠20只,随机分为卵巢切除(ovariectomized,OVX)组和假手术(sham operation,Sham)组,每组10只,分别进行卵巢去势手术和假手术处理。术后2、4、6、8周对两组大鼠进行活体显微CT扫描,测量股骨远端骨密度及相关松质骨形态计量学动态参数;同时经内眦静脉取血,利用蛋白芯片检测血清中不同蛋白因子的含量。结果显微CT检测结果显示OVX组大鼠骨密度(BMD)、相对骨体积(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)从第4周开始下降,骨小梁分离度(Tb.Sp)开始上升,Sham组未见明显变化;到第8周各指标在OVX组与Sham组之间差异有统计学意义(P<0.05)。蛋白芯片检测结果显示OVX组中干扰素-γ(IFN-γ)、β神经生长因子(b-NGF)分别从术后4周和6周后开始升高,同Sham组比较均至第8周具有明显差异(P<0.05)。结论 IFN-γ和b-NGF水平在绝经后骨质疏松早期开始增高,可考虑作为诊断早期绝经后骨质疏松症发生的新型蛋白分子。

Objective To screen the serum markers which can reflect the development of early stageof postmenopausal osteoporosis through protein chip. Methods Twenty 3-month-old female SD rats were ran-domly divided into sham operation control group（Sham） and ovariectomy group（OVX）, n=10 each. The bonemineral density（BMD） of distal femur and dynamic parameters of cancellous bone morphology of the two groupswere measured by micro CT at the end of 2, 4, 6, 8 weeks after operation. The blood of the two groups was takenfrom angular vein at the same time. The protein microarray was used to detect the concentration of 27 proteinfactors. Results Micro-CT examination showed that BMD, BV/TV, Tb.Th, Tb.Sp in OVX group began to de-crease and Tb.N began to increase at the end of 4 weeks, but there were no obvious changes in Sham group.There was significant difference between OVX and Sham groups by 8th weeks（P〈0.05）. Through the detectionof protein chip we found that IFN-gamma in OVX group increased at 4th week and b-NGF increased at 6thweek after operation, and there was significant difference between OVX and Sham groups by 8th weeks（P〈0.05）. Conclusion The concentrations of IFN-gamma and b-NGF began to increase at the early stage of post-menopausal osteoporosis. Both of them are likely to be used as new type of molecular markers in the diagnosis ofthe early stage of postmenopausal osteoporosis.