目的 观察三七皂苷单体(Notoginsenoside R1,NGR1)和成骨细胞之间的相关性.方法 将成骨前体细胞MC3T3-E1培养于含有10% FBS(GibcoTMInvitrogen)的α-MEM培养基中,通过在各培养孔中加入不同体积质量的三七皂苷单体来研究体外三七皂苷单体对成骨细胞的影响,检测方法为:荧光定量法检测细胞增殖情况;比色法检测细胞成骨分化的ALP活性情况;ELISA对各组OCN含量进行检测;RT-PCR检测,将β-actin表达作为内参照依据,校准各基因的临床表达水平情况.结果 同一时间点各体积质量的三七皂苷单体均呈现开口向下的抛物线状作用趋势,即在50 mg/L时显著促进细胞增殖,随后效果下降,超过100 mg/L则抑制细胞增殖(P<0.05);体积质量在50 mg/L时三七皂苷单体可显著促进ALP活性,随着体积质量的升高达到峰值后下降;早期三七皂苷单体上调OCN表达并无显著差异,7 d后其上调OCN表达量呈现体积质量依赖效应递增趋势;相关基因表达方面,三七皂苷单体在体积质量为50 mg/L时,对Runx2和OCN的基因表达均有较为明显促进作用,达到平台期后促进作用趋于稳定.结论 从成骨细胞增殖数量、碱性磷酸酶活性、骨钙素表达量等方面证实了一定体积质量的三七总皂苷单体对成骨细胞增殖的促进作用.%Objective To observe the relationship between Notoginsenoside R1(NGR1)and osteoblasts. Methods The osteoblastic progenitor cells MC3T3-E1 were cultivated in alpha-MEM culture medium containing 10%FBS(GibcoTMInvitrogen).Different concen-trations and different kinds of NGR1 were added for searching the effect of NGR1 on osteoblasts in vitro.The following were measured in this experiment:quantitative fluorescence methods were used to detect cell proliferation; cell osteogenic differentiation alkaline phos-phatase(ALP)activity was detected by colorimetric;OCN content in the medium was assayed through enzyme-linked immunosorbent assay(ELISA). The expression level of each gene was calibrated through real-time fluorescence quantitative RT-PCR detection, and the expression level of housekeeping beta -actin gene was taken as an internal reference. Results When the concentration of NGR1 was 50 mg/L,it could significantly promote cell proliferation,but the effect of increasing concentration decreased,more than 100 had the function of inhibiting cell proliferation;when the concentration of NGR1 was 50 mg/L,it could significantly promote the activity of ALP,with the increasing of NGR1 concentration,the activity of ALP increased,and reached a plateau after the fall. NGR1 up-regula-ted early,the expression of OCN was not significantly different,the expression of OCN increased when the concentration of NGR1(5-200 mg/L)was up-regulated after 7 days. There was no significant difference in the overall expression on the 7th day,however on the 7th day at the concentration of 200 mg/L the most significant effect was induced;NGR1 significantly promoted the proliferation of oste-oblasts,and gradually increased with the increasing of the concentration of NGR1. Conclusion NGR1 at fixed concentrations can pro-mote osteoblasts proliferation.
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