司坦唑醇对体外培养雌激素受抑的青春期大鼠生长板软骨细胞胶原合成的影响

Effects of stanozolol on collagen synthesization of cultured growth plate chondrocytes from pubertal female rat treated with GnRHa in vitro

目的探讨司坦唑醇（stanozolol，ST）对离体培养的促性腺激素释放激素拟似物（gonadotropin releasing hormone agonist，GnRHa）处理后青春期大鼠生长板软骨细胞的Ⅱ、Ⅹ型胶原合成的影响。方法将6只经GnRHa处理后的雌鼠原代软骨细胞分为时效组（观察ST干预时限的影响）、量效组（观察ST干预剂量的影响）。采用免疫组化法检测ST对软骨细胞Ⅱ、Ⅹ型胶原的表达。结果ST作用3d后胶原Ⅱ的表达明显高于对照组（P〈0．05），但至第5天却表达下降；而3d内胶原X分泌量与对照组比较无差异，4天起显著递增（P=0．000）。ST各个浓度的软骨细胞胶原Ⅱ表达较基值均明显增加（P〈0．01），并以10^-9M最显；10^-11M～10^-8M的ST不增加胶原X表达，至10^-7M始随剂量增加而增，10^-5M时达最高值（P〈0．01）。结论ST对Ⅱ、Ⅹ胶原的量效和时效影响存在“错峰”改变，时效影响的差异较量效差异为显著。对保持细胞生长／成熟正平衡，时效影响更需关注。

Objective To observe the effects of stanozolol on collagen Ⅱ and Ⅹ synthesization of cultured growth plate chondrocytes from pubertal female rat treated with GnRHa in vitro. Methods Primary Chondrocytes from 6 female rats were divided into two groups （ time-effect groups and concentration-effect groups）. immunohistochemical staining of collagen Ⅱ, Ⅹ were conducted. Results the results of type Ⅱ collagen synthesization demonstrated stanozolol enhanced the proliferation of the chondrocytes, time-course studies had shown that the proliferation were maximally stimulated by stanozolol after 3 days of incubation and decreased again after longer periods of incubation. Stanozolol did not stimulated type Ⅹ collagen synthesization from 10^-11 M^-10^-8M, but experiments showed that type Ⅹ collagen was already stimulated after incubation in stanozolol （10^-7-10^-5 M）. Time-course studies had shown those type Ⅹ collagen synthesizations were stimulated by stanozolol after 4-5 days of incubation. Conclusion Stanozolol enhances the proliferation of chondrocytes of pubertal female rat treated with GnRHa in vitro（time-course-dependent and concentration-dependent）.