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目的 研究四环素 哌嗪雌酚酮 (XW 6 30 )对长骨原癌基因c myc蛋白表达的影响 ,探讨其对骨的作用机制。方法 取 16d孕龄的雌性胚胎小鼠 ,剥取前肢尺骨 ,在特制的培养装置上于BGbJ培养基中培养4 8h ,培养基中XW6 30终浓度分别为 1× 10 -7,1× 10 -8和 1× 10 -9mol·L-1。用免疫组织化学方法测定长骨骺板c Myc蛋白的表达。图象分析系统下测定静止区、增殖区、肥大区c Myc蛋白免疫反应阳性细胞面积。结果 当培养基中XW6 30浓度为 1× 10 -9mol·L-1时 ,增殖区阳性细胞面积与同浓度雌酚酮组相比增加 ,静止区阳性细胞面积与对照组相比也增加 ,此浓度的雌酚酮组静止区阳性细胞面积与对照组无差异。XW 6 30浓度为 1× 10 -8和 1× 10 -7mol·L-1时 ,各区阳性细胞面积均比对照组及同浓度雌酚酮组增加。结论 XW6 30对骨的雌激素活性强于雌酚酮。XW 6 30可能通过促进骺板软骨细胞表达c Myc蛋白而促进了软骨细胞的增殖和分化 ,从而促进了软骨内成骨。
Objective To study the effect of tetracycline piperazine estrone (XW 6 30) on the expression of c-myc protein in long bone and explore its mechanism of action on bone. Methods Female embryonic mice of gestational age of 16 days were harvested and the ulna of forelimb was stripped and cultured on BGbJ medium for 48 h. The final concentration of XW6 30 in culture medium was 1 × 10 -7 and 1 × 10 - 8 and 1 × 10 -9 mol·L-1. Immunohistochemistry was used to detect the expression of c Myc in the long bone. The image analysis system was used to determine the area of c Myc protein immunoreactive cells in quiescent zone, proliferative zone and hypertrophic zone. Results When the concentration of XW6 30 in culture medium was 1 × 10 -9 mol·L-1, the area of positive cells in proliferating zone increased compared with that in the same concentration of estrone, and the area of positive cells in quiescent zone also increased compared with the control group Concentration of estrone group quiescent zone positive cell area and the control group no difference. When the concentration of XW 6 30 was 1 × 10 -8 and 1 × 10 -7 mol·L-1, the area of positive cells in each area increased compared with the control group and the same concentration of estrone group. Conclusion The estrogen activity of bone in XW6 30 is stronger than that of estrone. XW 6 30 may promote chondrogenic osteogenesis through promoting the proliferation and differentiation of chondrocytes by promoting the expression of c Myc protein in epiphyseal plate chondrocytes.