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目的:探讨α-玉米赤霉醇(α-zearalanol,α-ZAL)对小鼠成骨样细胞MC3T3-E1增殖、分化及护骨素(osteoprotegerin,OPG)和NF-кB受体活化配体(receptor activator of nuclear factor-кB ligand,RANKL)mRNA表达的影响。方法:传代培养小鼠成骨样细胞株MC3T3-E1,采用不同浓度的α-ZAL作用于细胞72 h后,MTT法检测细胞增殖活性,PNPP法检测碱性磷酸酶(alkaline phosphatase,ALP)活性,RT-PCR法检测ALP,OPG及RANKL mRNA的表达水平。结果:10-6～10-12mol·L-1的α-ZAL可显著抑制成骨细胞的增殖(P<0.05);显著增加ALP活性(P<0.05),但不同剂量间存在作用时间差异;并且可显著上调成骨细胞内OPG/RANKL mRNA的比值(P<0.05)。结论:α-ZAL可抑制成骨细胞的增殖、促进其分化,并可通过上调OPG/RANKL mRNA表达比值抑制破骨细胞的形成,有望作为骨质疏松症的治疗药物。
OBJECTIVE: To investigate the effects of α-zearalanol (α-ZAL) on the proliferation and differentiation of osteoblast-like cells MC3T3-E1 and the activation of osteoprotegerin (OPG) and NF- receptor activator of nuclear factor-кB ligand, RANKL) mRNA expression. Methods: The osteoblast-like cell line MC3T3-E1 was subcultured. The cells were treated with different concentrations of α-ZAL for 72 h. The cell proliferation activity was detected by MTT assay. The activity of alkaline phosphatase (ALP) The mRNA expression of ALP, OPG and RANKL were detected by RT-PCR. Results: α-ZAL at 10-6-10-12 mol·L-1 could significantly inhibit the proliferation of osteoblasts (P <0.05) and increase the activity of ALP significantly (P <0.05), but there was a time-effect difference between different doses of α-ZAL. And up-regulated the ratio of OPG / RANKL mRNA in osteoblasts (P <0.05). CONCLUSION: α-ZAL can inhibit the proliferation and promote the differentiation of osteoblasts and inhibit the formation of osteoclasts by up-regulating the ratio of OPG / RANKL mRNA expression. It is expected to be used as a therapeutic drug for osteoporosis.