骨质疏松肾阳虚和肾阴虚证型下左归丸含药血清干预成骨细胞ERK12,Wntβcatenin信号通路研究.docVIP

骨质疏松肾阳虚和肾阴虚证型下左归丸含药血清干预成骨细胞ERK12,Wntβcatenin信号通路研究 　　[摘要]?文研究骨质疏松（OP）肾阳虚和肾阴虚证型下左归丸含药血清对成骨细胞增殖和碱性磷酸酶（ALP）表达及对成骨细胞ERK1/2，Wnt/βcatenin信号通路相关因子βcatenin，ERK1，ERK2 mRNA及蛋白表达的作用。采用手术去卵巢法造成大鼠骨质疏松模型，并在手术10周后，分别注射氢化可的松、灌胃甲状腺素造成大鼠OP肾阳虚及OP肾阴虚模型；用新生24 h乳鼠颅骨体外培养得成骨细胞，以碱性磷酸酶和茜素红染色进行鉴定，分别用OP肾阳虚、OP肾阴虚及OP组（对照组）所得的左归丸含药血清、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，RTPCR法检测ERK1/2，βcatenin mRNA的表达，Western blot法检测ERK1/2，βcatenin蛋白表达。结果表明：OP肾阴虚证型下左归丸含药血清在促进成骨细胞增殖、ALP表达、成骨细胞ERK1/2，Wnt/βcatenin信号通路相关因子βcatenin，ERK1，ERK2 mRNA及蛋白表达方面比OP肾阳虚证型下左归丸含药血清的作用更强。这与“左归丸滋肾阴”的中医理论相吻合，为临床辩证治疗骨质疏松症提供科学依据。左归丸通过ERK1/2和Wnt/βcatenin信号通路调控成骨细胞的增殖与分化，可能是左归丸防治骨质疏松肾阴虚证的机制之一。 　　[关键词]左归丸； 成骨细胞； 含药血清； 骨质疏松肾阳虚； 骨质疏松肾阴虚； Wnt/βcatenin信号通路； ERK1/2信号通路 　　[Abstract]To clarify the effects of Zuoguiwan containing serum on osteoblast proliferation and alkaline phosphatase（ALP） expression and its effects on the expression of βcatenin， ERK1， ERK2 mRNA and protein of osteoblast through ERK1/2， Wnt/βcatenin signaling pathway in models with osteoporosis（OP） kidneyYangdeficiency， osteoporosis（OP） kidneyYindeficiency syndrome Rat osteoporosis models were established by ovariectomy surgery， and 10 weeks after surgery， hydrocortisone was injected and thyroxine was administered by intragastric administration to establish OP kidneyYangdeficiency rat model， and OP kidneyYindeficiency rat model Osteoblasts were obtained from 24 h newborn rat skull and were identified by alkaline phosphatase and alizarin red staining Zuoguiwan containing serum of OP， OP kidneyYangdeficiency， and OP kidneyYindeficiency， as well as the blank serum were used to intervene the osteoblast， and the cells proliferation was detected by MTS ELISA assay was used to detect ALP expression RTPCR assay was used to detect the mRNA expression of ERK1， ERK2， βcatenin and protein expression levels were detected by Western blot The results showed that Zuoguiwan containing serum in OP kidneyYindeficiency model had stronger effect than OP kidneyYangdeficiency in promoting ost