骨质疏松肾阳虚、肾阴虚证型下右归丸含药血清对大鼠成骨细胞ERK1/2、Wnt/β-catenin信号通路的影响

Study on the effects of Yougui Pills containing serum on osteoblasts ERK1/2 and Wnt/β-catenin signaling pathway with syndrome of kidney-yang deficiency and kidney-yin deficiency osteoporosis

目的:研究骨质疏松症(OP)肾阳虚和肾阴虚证型下右归丸含药血清对大鼠成骨细胞增殖和碱性磷酸酶(ALP)表达的作用,及对成骨细胞ERK1/2、Wnt/β-catenin信号通路相关因子β-catenin、ERK1、ERK2mRNA及蛋白表达的作用。方法:分别用OP肾阳虚、OP肾阴虚及OP组(对照组)所得的右归丸含药血清及空白血清干预成骨细胞,采用MTS法检测细胞增殖情况,ELISA法检测ALP表达,RT-PCR法检测ERK1、ERK2、β-catenin mRNA的表达,Western Blot法检测ERK1、ERK2、β-catenin蛋白的表达。结果:OP肾阳虚证型下右归丸含药血清在促进成骨细胞增殖、ALP表达以及对成骨细胞ERK1/2、Wnt/β-catenin信号通路相关因子β-catenin、ERK1、ERK2 mRNA及蛋白表达方面比OP肾阴虚证型下右归丸含药血清的作用更强。结论:与"右归丸补肾阳"的中医理论相吻合,为临床辨证治疗OP提供科学依据。

Objective： To study the effects of Yougui Pills containing serum on osteoblasts proliferation and alkaline phosphatase（ALP） expression under osteoporosis（OP） with deficiency of kidney yang and deficiency of kidney yin model, and the effects on osteoblast ERK1/2, Wnt/β-catenin signaling pathway related genes factors β-catenin, ERK1, ERK2 mRNA and protein expressions. Methods： The osteoblasts were induced with the Yougui Pills containing serum of OP kidney-yang deficiency, OP kidney-yin deficiency and OP（control group） respectively. The proliferation of osteoblasts was detected by MTS assay. The expression of ALP was detected by ELISA. The mRNA expressions of ERK1, ERK2 and β-catenin were detected by RT-PCR. The protein expressions of ERK1, ERK2 and β-catenin were detected by Western Blot. Results： Yougui Pills containing serum of OP kidney-yang deficiency can promote the proliferation and expression of ALP, the mRNA and protein expressions of ERK1/2, Wnt/β-catenin signaling pathway related factors β-catenin, ERK1, ERK2 more significant than those of Yougui Pills containing serum of OP kidney-yin deficiency. Conclusion： Yougui Pills is more suitable for f OP kidney-yang deficiency, which is consistent with the TCM theory that ‘Yougui Pills is for kidney yang deficiency＇, and provides a scientific basis for clinical syndrome differentiation and treatment of osteoporosis.