咪达唑仑抑制人骨髓间充质干细胞的成骨分化。

Midazolam suppresses osteogenic differentiation of human bone marrow-derived mesenchymal stem cells.

作者信息

Zhang T, Shao H, Xu K-q, Kuang L-t, Chen R-f, Xiu H-h

机构信息

Department of Anesthesiology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2014;18(9):1411-8.

PMID:24867522

Abstract

OBJECTIVE

Previous study showed that peripheral-type benzodiazepine receptors (PBRs) are expressed in human mesenchymal stem cells (hMSCs) and diazepam was found to inhibit hMCSs viability in high concentration. Midazolam, a benzodiazepine derivative, is widely used as an intravenous sedative in hospital. Peripheral-type benzodiazepine receptors (PBRs) affect a broad spectrum of cellular functions. We tested the cell viability and osteogenic differentiation of hMSCs.

PATIENTS AND METHODS

Bone marrow was collected from 12 patients during the operation of spine internal fixation. Cultivated with basal medium, the hBMSCs were incubated with or without midazolam (0.1, 1, 5, 10, 15, 20 µM, respectively). Cell viability were tested with MTS assay after 2, 4, 6 hours respectively. Cell morphology was observed and recorded at 6 hour. After cultivated with osteogentic medium, the hBMSCs were incubated with or without midazolam (5, 10, 15, 20 µM, respectively). Alkaline phosphatase (ALP) activity and alizarin red S staining were measured. Cultivated with osteogentic medium with or without treatment of 15 µM midazolam, the mRNA expression of ALP, type 1 collagen (COL1), Runx2 and PPARγ was analyzed by real-time RT-PCR.

RESULTS

The treatments of midazolam inhibited cell viability to 85%-16% respectively (p < 0.05). Rounded up phenomenon with floating cells, Membrane-blebbed cells and cytoplasmic contraction were observed after 10, 15 or 20 µM midazolam treatment. The ALP activity and Calcium deposition of hBMSCs exposed to 15 and 20 µM midazolam was significantly inhibited at 7, 14 and 21 days (p < 0.05). And the mRNA expression of ALP, COL1 and PPARγ was significantly suppressed in the hBMSCs cultured with 15 µM midazolam (p < 0.05).

CONCLUSIONS

Midazolam exert negative effect on cell viability and osteogenic differentiation of cultured hBMSCs. During sedation in critical care, the use of midazolam may suppress activity of hBMSCs.

摘要

目的

先前的研究表明，外周型苯二氮䓬受体（PBRs）在人间充质干细胞（hMSCs）中表达，并且发现高浓度地西泮会抑制hMCSs的活力。咪达唑仑作为一种苯二氮䓬衍生物，在医院中广泛用作静脉镇静剂。外周型苯二氮䓬受体（PBRs）影响广泛的细胞功能。我们测试了hMSCs的细胞活力和成骨分化。

患者和方法

在脊柱内固定手术期间从12名患者采集骨髓。用基础培养基培养，hBMSCs分别在有或没有咪达唑仑（分别为0.1、1、5、10、15、20μM）的情况下孵育。分别在2、4、6小时后用MTS法测试细胞活力。在6小时时观察并记录细胞形态。在用成骨培养基培养后，hBMSCs分别在有或没有咪达唑仑（分别为5、10、15、20μM）的情况下孵育。测量碱性磷酸酶（ALP）活性和茜素红S染色。在用15μM咪达唑仑处理或未处理的成骨培养基中培养后，通过实时RT-PCR分析ALP、1型胶原（COL1）、Runx2和PPARγ的mRNA表达。

结果

咪达唑仑处理分别将细胞活力抑制至85%-16%（p<0.05）。在10、15或20μM咪达唑仑处理后观察到细胞出现圆缩现象、有漂浮细胞、细胞膜泡状化细胞和细胞质收缩。在7、14和21天时，暴露于15和20μM咪达唑仑的hBMSCs的ALP活性和钙沉积受到显著抑制（p<0.05）。并且在15μM咪达唑仑培养的hBMSCs中，ALP、COL1和PPARγ的mRNA表达受到显著抑制（p<0.05）。