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该文研究了不同浓度刺老苞根皮含药血清对原代成骨细胞Wnt/β-catenin信号通路中β-catenin,Wnt-1,Frizzed-2,TCF和Axin表达的影响。经SPF级健康雌性SD大鼠(80只)给予蒸馏水、刺老苞根皮水煎剂高、中、低3种剂量,连续灌胃7d,制备刺老苞根皮含药血清。体外培养原代成骨细胞(OB)并鉴定,取第3代成骨细胞,培养48 h后用各组含药血清干预培养10 d,采用茜素红染色钙化结节并计数;干预培养48 h后收集细胞,分别采用Real-time PCR(RT-PCR)和Western blot法测定β-catenin,Wnt-1,Frizzed-2,TCF和Axin的表达情况。结果显示,经鉴定体外培养细胞为原代成骨细胞;在干预培养后,与模型组相比,各剂量组可促进原代成骨细胞的矿化能力,且上调β-catenin,Wnt-1,Frizzed-2,TCF mRNA和蛋白的表达,下调Axin mRNA和蛋白的表达。结果发现刺老苞根皮含药血清可通过调节原代成骨细胞Wnt/β-catenin信号通路中β-catenin,Wnt-1,Frizzed-2,TCF和Axin表达,增强成骨细胞的分化和增殖。
The aim of this study was to investigate the effect of different concentrations of drug-containing serum from root bark on the expression of β-catenin, Wnt-1, Frizzed-2, TCF and Axin in primary osteoblast Wnt / β-catenin signaling pathway. SPF healthy female SD rats (80 rats) were given distilled water, the old Bupleurum root decoction decoction of high, medium and low doses of 3, continuous gavage 7d, preparation of thorn old bract root bark containing serum. Primary osteoblasts (OBs) were cultured in vitro and identified. The third generation osteoblasts were harvested for 48 h. The osteoblasts were harvested for 10 days. The calcified nodules were stained with alizarin red and counted. h. The cells were collected and the expressions of β-catenin, Wnt-1, Frizzed-2, TCF and Axin were detected by Real-time PCR and Western blot respectively. The results showed that the cultured cells were identified as primary osteoblasts. Compared with the model group, each dose group could promote the mineralization ability of primary osteoblasts and up-regulate the expression of β-catenin, Wnt-1 , Frizzed-2, TCF mRNA and protein expression, down-regulated Axin mRNA and protein expression. The results showed that the drug-containing serum of Eupatorium majus could enhance the differentiation of osteoblasts by regulating the expression of β-catenin, Wnt-1, Frizzed-2, TCF and Axin in Wnt / β-catenin signaling pathway of primary osteoblasts proliferation.