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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 dynamic changes of bone marrow-derived osteoclasts and the changes of osteoprotegerin (OPG) gene in bone marrow of castrated rats. Methods Sixty healthy Wistar female rats were divided into experimental group (ovariectomized group) and control group (sham operated group), 30 in each group. The ovariectomized group was taken at 2, 4, 6, 8 and 12 weeks And control group, each rat femur bone marrow cell culture, the left femur culture cells were Wright - Giemsa staining and tartrate acid phosphatase (TRAP) staining observed osteoclasts and count. Total RNA was extracted from the right femur culture cells and the expression of OPG mRNA in ovariectomized rat bone marrow cells was observed by RT-PCR. Results The number of osteoclasts in castration group was more than that in control group at 2 and 4 weeks after operation. The formation of osteoclasts peaked at 6 weeks in castration group, which was significantly more than that in control group Cell number decreased compared with 6 weeks, but still significantly higher than the control group. OPG mRNA expression in bone marrow cells cultured at 2 and 4 weeks after OPG mRNA expression compared with the control group had no significant difference, 6 weeks castration group OPG mRNA expression was significantly lower than the control group, 8 weeks castration group OPG mRNA expression compared with the control group No significant difference. Conclusion Ovariectomy can result in the increase of osteoclast formation and the decrease of OPG mRNA expression in both groups, all at 6 weeks after operation.