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补肾益髓类中药含药血清对骨髓基质细胞增殖及成骨分化的影响

Effect of kidney-reinforcing and marrow-beneficial traditional Chinese medicine-intervened serum on the proliferation and osteogenic differentiation of bone marrow stromal cells.

作者信息

Zhou DA-An, Deng Yue-Ning, Liu Lei, Li Jian-Jun

机构信息

Department of Spinal and Neural Function Reconstruction, China Rehabilitation Research Center, School of Rehabilitation Medicine of Capital Medical University, Beijing 100068, P.R. China ; Department of Rehabilitation, The Third Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121000, P.R. China.

Department of Rehabilitation, The Third Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121000, P.R. China.

出版信息

Exp Ther Med. 2015 Jan;9(1):191-196. doi: 10.3892/etm.2014.2062. Epub 2014 Nov 11.

Abstract

The present study aimed to investigate the effect of kidney-reinforcing and marrow-beneficial traditional Chinese medicine (TCM)-intervened (KRMBTI)-serum on the proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs) in rats. Rat BMSCs were isolated and cultured with various concentrations of serum obtained from rats at different time-points following treatment with low, medium and high doses of KRMBT. The alkaline phosphatase (ALP) activity and proliferation of the BMCSs was assessed to determine the optimal serum sampling time-point and serum concentration. Transforming growth factor (TGF)-β1 expression of the BMSCs was detected using enzyme-linked immunosorbent assay (ELISA), and hepcidin mRNA expression in the rat livers was detected using reverse transcription polymerase chain reaction. The proliferation of BMCSs treated with serum obtained l h after dosing was observed to be significantly higher than that for BMCSs treated with serum obtained at the four other time-points (P<0.05). Furthermore, the proliferation following treatment with 25% KRMBTI-serum was significantly higher than that for the other KRMBTI-serum concentrations (P<0.01). For a 25% concentration of the serum collected at l h, the proliferation in the high- and low-dose KRMBTI-serum groups was significantly higher than that of the medium-dose and control groups (P<0.01) and no statistical significance was observed between the high- and low-dose groups. In the osteogenic differentiation process of the high-dose group, the ALP activity at every time-point was significantly higher than that of the low-dose group and the peak value of the former was achieved at concentrations between 20 and 30%. KRMBTI-serum was shown to promote the expression of TGF-β1. Furthermore, hepcidin was observed to be expressed at significantly higher levels in the high-dose group than in the control group, and hepcidin expression was significantly higher after 10 weeks compared with that after five weeks. These findings suggest that KRMBTI-serum increases TGF-β1 and hepcidin expression levels, which may be the mechanism underlying the promotion of osteogenic differentiation induced by KRMBTI-serum in BMSCs.

摘要

本研究旨在探讨补肾益髓中药干预血清(KRMBTI)对大鼠骨髓基质细胞(BMSCs)增殖和成骨分化的影响。分离培养大鼠BMSCs,用低、中、高剂量KRMBT处理大鼠后在不同时间点获取的不同浓度血清培养BMSCs。评估骨髓基质细胞的碱性磷酸酶(ALP)活性和增殖情况,以确定最佳血清采样时间点和血清浓度。采用酶联免疫吸附测定(ELISA)检测BMSCs中转化生长因子(TGF)-β1的表达,采用逆转录聚合酶链反应检测大鼠肝脏中铁调素mRNA的表达。给药后1小时获取的血清处理的骨髓基质细胞的增殖明显高于其他四个时间点获取的血清处理的骨髓基质细胞(P<0.05)。此外,25%KRMBTI血清处理后的增殖明显高于其他KRMBTI血清浓度(P<0.01)。对于1小时采集的25%浓度血清,高剂量和低剂量KRMBTI血清组的增殖明显高于中剂量组和对照组(P<0.01),高剂量组和低剂量组之间无统计学意义。在高剂量组的成骨分化过程中,各时间点的ALP活性明显高于低剂量组,前者在20%至30%浓度时达到峰值。结果表明KRMBTI血清可促进TGF-β1的表达。此外,高剂量组铁调素的表达明显高于对照组,且10周后的铁调素表达明显高于5周后。这些研究结果表明,KRMBTI血清可提高TGF-β1和铁调素的表达水平,这可能是KRMBTI血清促进BMSCs成骨分化的潜在机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/552c/4247301/7b6c4cf17485/ETM-09-01-0191-g00.jpg

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