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目的 研究孕激素对正常成人成骨细胞转化生长因子 (TGF) β1、TGF β2表达的调节作用 ,探讨孕激素治疗绝经后骨质疏松症 (OP)的作用机制。方法 鉴定成人成骨细胞 (hOB) ,测定碱性磷酸酶 (ALP)活性、骨钙素分泌 ,VanGieson氏苦味酸酸性复红染色法进行Ⅰ型胶原染色。hOB用孕酮干预。Northern杂交、ELISA分别检测hOBTGF β1、TGF β2mRNA和蛋白质分泌。结果 观察到hOB分泌的ALP活性为 (74 3± 4 7)mU/mg蛋白 ,培养上清液骨钙素含量为 (3 84± 0 3 9)ng/ml蛋白 ,Ⅰ型胶原染色呈红色。观察到孕酮增强hOBTGF β1、TGF β2mRNA表达呈剂量依赖性 (P <0 0 0 1) ;10 -9mol/L孕酮诱导 12～ 2 4h促进TGF β1、TGF β2mRNA表达 ,呈时间依从性。孕酮促进hOBTGF β1、TGF β2蛋白质分泌呈剂量依赖性 (P <0 0 0 1) ;10 -9mol/L孕酮诱导 12～ 2 4h促进TGF β1、TGF β2蛋白质分泌 ,呈时间依从性。结论 孕激素可能通过诱导成骨细胞TGF β1、TGF β2表达 ,促进成骨细胞增殖与分化 ,增强成骨功能及骨基质合成 ,促进骨形成
Objective To study the regulatory effect of progesterone on the expression of transforming growth factor β1 and TGF β2 in adult human osteoblasts and to explore the mechanism of action of progesterone in the treatment of postmenopausal osteoporosis (OP). Methods Adult osteoblasts (hOB) were identified and alkaline phosphatase (ALP) activity, osteocalcin secretion and VanGieson’s picric acid complex complex red staining were used to stain type Ⅰ collagen. hOB with progesterone intervention. Northern blot and ELISA were used to detect hOBTGF β1, TGF β2 mRNA and protein secretion respectively. Results The ALP activity of hOB was (74 3 ± 4 7) mU / mg protein, the content of osteocalcin in culture supernatant was (3 84 ± 0 9 9) ng / ml protein, and type Ⅰ collagen was red. It was observed that progesterone enhanced the expression of hOBTGFβ1 and TGFβ2 mRNA in a dose-dependent manner (P <0.01). When induced by 10 -9 mol / L progesterone, the expression of TGFβ1 and TGFβ2 mRNA was promoted in a time-dependent manner. Progesterone promoted the secretion of hOBTGFβ1 and TGFβ2 in a dose-dependent manner (P <0.01). When induced by 10 -9mol / L progesterone, the secretion of TGFβ1 and TGFβ2 increased in a time-dependent manner. Conclusion Progesterone may promote the proliferation and differentiation of osteoblasts by inducing the expression of TGF-β1 and TGF-β2 in osteoblasts, enhance osteogenesis and bone matrix synthesis, and promote bone formation