贝尼地平对处于细胞周期各阶段的培养人系膜细胞具有广泛的抗增殖作用。

Broad antiproliferative effects of benidipine on cultured human mesangial cells in cell cycle phases.

作者信息

Ono Takahiko, Liu Ning, Kusano Hitoshi, Nogaki Fumiaki, Makino Toshiaki, Muso Eri, Sasayama Shigetake

机构信息

Nephrology Division, Department of Cardiovascular Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan.

出版信息

Am J Nephrol. 2002 Sep-Dec;22(5-6):581-6. doi: 10.1159/000065266.

DOI:10.1159/000065266

PMID:12381964

Abstract

BACKGROUND AND PURPOSE

Recently, in vitro studies have shown that some calcium channel blockers inhibit the proliferation of mesangial cells. In the present study, we evaluated the antiproliferative effects of benidipine, a calcium channel blocker, in comparison with other calcium channel blockers, and attempted to further clarify its mechanism of action on cultured human mesangial cells in relation to cell cycle.

METHODS

Human mesangial cells were cultured in medium containing 5% fetal calf serum (FCS), with or without benidipine, or other calcium channel blockers for 20 h, and [(3)H]thymidine incorporation were measured. Analysis of cell cycle dependency was carried out, using platelet-derived growth factor as a competence factor, which transfers cells from the G0 to the G1 (G0/G1) phase, and insulin as a progression factor, which transfers cells from the G1 to the S (G1/S) phase. Cells were also analyzed by flow cytometry.

RESULTS

Benidipine and nifedipine showed significant inhibitory effects on FCS-induced proliferation (p < 0.001 and p < 0.01, respectively, by ANOVA), with 3.8 and 41.9% of the control level of [(3)H]thymidine incorporation at the concentration of 10 microM of the blockers, respectively. Diltiazem had no inhibitory effect at this concentration. Benidipine was found to inhibit cells in both the boundaries of G0/G1 and G1/S phases (p < 0.001 and p < 0.0001, respectively), whereas nifedipine inhibited only cells in G1/S phase (p < 0.05). The effects of benidipine (10 microM) on cells in G1/S were stronger than those on cells in the G0/G1 phase (p < 0.0001). Furthermore, flow cytometry showed that 10 mM benidipine significantly inhibited the G1 to S phase transition of FCS-stimulated mesangial cells (p < 0.03).

CONCLUSIONS

Benidipine markedly inhibited the proliferation of mesangial cells, at both the G0/G1 and G1/S phases, and it might be effective to suppress the progression of mesangioproliferative glomerular diseases.

摘要

背景与目的

最近，体外研究表明一些钙通道阻滞剂可抑制系膜细胞的增殖。在本研究中，我们评估了钙通道阻滞剂贝尼地平与其他钙通道阻滞剂相比的抗增殖作用，并试图进一步阐明其对培养的人系膜细胞作用的机制及其与细胞周期的关系。

方法

将人系膜细胞培养于含5%胎牛血清（FCS）的培养基中，分别加入或不加入贝尼地平或其他钙通道阻滞剂培养20小时，然后检测[³H]胸腺嘧啶核苷掺入量。以血小板衍生生长因子作为启动因子（可使细胞从G0期进入G1期）和胰岛素作为进展因子（可使细胞从G1期进入S期），进行细胞周期依赖性分析。同时采用流式细胞术对细胞进行分析。

结果

贝尼地平和硝苯地平对FCS诱导的增殖具有显著抑制作用（方差分析分别为p < 0.001和p < 0.01），在阻滞剂浓度为10微摩尔时，[³H]胸腺嘧啶核苷掺入量分别为对照水平的3.8%和41.9%。地尔硫䓬在该浓度下无抑制作用。发现贝尼地平可抑制处于G0/G1期和G1/S期边界的细胞（分别为p < 0.001和p < 0.0001），而硝苯地平仅抑制G1/S期的细胞（p < 0.05）。贝尼地平（10微摩尔）对G1/S期细胞的作用强于对G0/G1期细胞的作用（p < 0.0001）。此外，流式细胞术显示10毫摩尔贝尼地平可显著抑制FCS刺激的系膜细胞从G1期向S期的转变（p < 0.03）。