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目的:研究车前子含药血清对大鼠颅骨成骨细胞体系增殖、分化及成骨能力的影响,并初步探讨上皮钠离子通道(ENaC)参与车前子调节成骨细胞功能的作用。方法:按血清药理学实验方法制备车前子含药血清和对照血清,采用酶消化法分离培养大鼠成骨细胞,CCK-8试剂盒和AKP试剂盒分别测定车前子含药血清对细胞增殖和分化的影响,半定量PCR方法测定车前子含药血清对成骨细胞ENaC和成骨功能指标基因表达的影响。结果:不同浓度的车前子含药血清均能显著刺激成骨细胞增殖、分化,其促进作用与药物浓度呈正相关关系,具有浓度依赖性;与对照组比较,低浓度车前子含药血清能显著上调成骨细胞α-ENaC基因的表达水平,其增加趋势与成骨功能指标基因(OC、ALP、OP)mRNA表达的增加趋势一致。结论:车前子含药血清不仅刺激成骨细胞增殖分化,在促进骨形成的同时也增加α-ENaC mRNA表达,提示其作用机理可能与ENaC介导的调节途径有关,为中药治疗骨代谢疾病的研究提供了一个新思路。
OBJECTIVE: To study the effect of psyllium containing serum on the proliferation, differentiation and osteogenic ability of osteoblasts in the skull of rats, and to explore the role of ENaC in the regulation of osteoblast function by psyllium. Methods: The serum and control serum of Psyllium were prepared according to the method of serum pharmacology. The osteoblasts were isolated and cultured by enzymatic digestion method. CCK-8 kit and AKP kit were used to measure the effect of Psyllium-containing serum on cells Proliferation and differentiation. The semi-quantitative PCR method was used to determine the effect of psyllium-containing serum on the expression of ENaC and osteogenic markers of osteoblasts. Results: Different concentrations of psyllium containing serum could significantly stimulate the proliferation and differentiation of osteoblasts, and its promoting effect was positively correlated with drug concentration, with a concentration-dependent manner. Compared with the control group, The expression of α-ENaC gene in osteoblasts was significantly up-regulated. The increasing tendency was consistent with the increasing trend of osteoblast function index genes (OC, ALP, OP) mRNA expression. CONCLUSION: Psyllium-containing serum not only stimulates the proliferation and differentiation of osteoblasts, but also enhances the expression of α-ENaC mRNA in promoting bone formation, suggesting that its mechanism of action may be related to the ENaC-mediated regulatory pathway, The research provides a new idea.