壮骨止痛胶囊调控OSX、OPN蛋白对去势大鼠成骨细胞分化的影响

目的 探究壮骨止痛胶囊含药血清调控成骨相关转录因子抗体（osterix，OSX）、骨桥蛋白（osteopontin，OPN）对去势大鼠成骨细胞分化的机制。方法 50只3月龄雌性大鼠，随机分为空白组、假手术组、模型组、骨松宝组和壮骨止痛胶囊组，10只/组，除空白组及假手术组外，摘除双侧完整卵巢，然后各组分别予以相应干预，获取相应血清；10只新生SD乳鼠提取原代成骨细胞，培养至第三代后添加上述分组相应10%血清，银染及碱性磷酸酶（alkaline phosphatase，ALP）染色鉴定成骨细胞，免疫组化检测OSX、OPN蛋白的表达。结果 各组中的成骨细胞银染呈褐色、碱性磷酸酶染色呈蓝紫色；免疫组化结果表明，骨松宝组、壮骨止痛胶囊组中细胞质呈棕色；空白组和假手术组呈浅棕黄色；模型组几乎不见棕黄色。与假手术组比，模型组OSX、OPN蛋白的表达率降低（P<0.05）；壮骨止痛方及骨松宝干预后OSX、OPN蛋白表达率较模型组显著升高，差异有统计学意义（P<0.05）。结论 壮骨止痛胶囊组含药血清能促进OSX、OPN蛋白分泌，从而促进成骨细胞增殖、分化，发挥抗绝经后骨质疏松作用。

Effect of Zhuanggu Zhitong Capsule on Osteoblast Differentiation in Ovariectomized Rats by Regulating Osterix and Osteopontin

Objective To investigate the effect of Zhuanggu Zhitong Capsule-containing serum on osteoblast differentiation in ovariectomized rats by regulating osterix (OSX) and osteopontin (OPN). Methods Fifty three-month-old female rats were equally and randomly divided into blank group, sham-operation group, model group, Gusongbao group, and Zhuanggu Zhitong Capsule group. Both intact ovaries were taken for all rats except those in the blank group and the sham-operation group. And then each group was given the corresponding intervention to collect their serum. Ten neonatal Sprague-Dawley mice were used to isolate primary osteoblasts. When the primary osteoblasts were cultured to the third generation, the 10% serum from the above groups was added. Silver staining and alkaline phosphatase (ALP) staining were used to identify osteoblasts, and immunohistochemistry was used to determine the expression of OSX and OPN. Results The osteoblasts in each group were brown in silver staining assay and blue-violet in ALP staining assay. The immunohistochemical results showed that the cytoplasm was brown in the Gusongbao group and the Zhuanggu Zhitong Capsule group; the cytoplasm was light brownish yellow in the blank group and the sham-operation group; no brownish yellow was observed in the model group. Compared with the sham-operation group, the model group had significantly lower expression rates of OSX and OPN (P<0.05). The Zhuanggu Zhitong Capsule group and the Gusongbao group had significantly higher expression rates of OSX and OPN than the model group (P<0.05). Conclusion The Zhuanggu Zhitong Capsule-containing serum can promote the secretion of OSX and OPN to promote the proliferation and differentiation of osteoblasts, thus exerting anti-menopausal osteoporosis.