去势大鼠骨髓体外培养破骨细胞观察及OPG mRNA表达的变化

Observation of bone marrow cells in ovariectomied rats and expression of osteoprotegerin mRNA

目的观察去势大鼠骨髓源性破骨细胞形成的动态变化和骨髓细胞护骨素（OPG）基因的变化。方法健康Wistar雌性大白鼠60只,分为实验组（去卵巢组）和对照组（假手术组）,每组30只,分别于术后2、4、6、8和12周取去卵巢组和对照组每只大鼠股骨骨髓细胞进行细胞培养,左侧股骨培养细胞作瑞氏-吉姆萨染色和抗酒石酸酸性磷酸酶（TRAP）染色,观察破骨细胞并计数。右侧股骨培养细胞提取总RNA,通过RT-RCR来观察去势大鼠骨髓细胞OPG mRNA表达强度变化。结果术后2、4周去势组破骨细胞形成数多于同期对照组,6周去势组破骨细胞形成达到高峰,显著多于同期对照组P〈0.01。8周去势组破骨细胞数较6周有所下降,但仍明显高于对照组。骨髓培养细胞OPG mRNA表达,术后2、4周OPG mRNA表达较同期对照组无明显差别,6周去势组OPG mRNA表达明显低于对照组,8周去势组OPG mRNA表达与同期对照组比较无明显差异。结论大鼠去势后可引起破骨细胞形成数量增多和OPG mRNA表达降低,且均在术后6周表现明显。

Objective To observe the changes of bone marrow osteoclast formation and expression of OPG gene. Methods 60 Wistar female rats were divided into experimental group （ovariectomied group） and control group （sham operating group）. The bone marrow cells were cultured and TRAP staining were performed. RT-PCR was performed to detect the expression of OPG mRNA in ovariectomied rats. Results The number of the osteoclast formation was more than that in the contrast group,and it reached the peak at the 6 th week after variectomy,which was significantly more than that in the contrast group（P〈0.01）. The expression of osteoprotegerin mRNA in ovariectomied group was obviously lower than that in the contrast group at the 6 th week. Conclusion The number of osteoclast formation in ovariectomied rats increased and expression of OPG mRNA decreased,which were all obvious at the 6 th week after variectomy.