洛伐他汀抑制培养的主动脉平滑肌细胞缝隙连接通讯。

Lovastatin inhibits gap junctional communication in cultured aortic smooth muscle cells.

机构信息

Department of Cardiovascular Sciences, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.

出版信息

J Cardiovasc Pharmacol Ther. 2010 Sep;15(3):296-302. doi: 10.1177/1074248410373750. Epub 2010 Jul 2.

DOI:10.1177/1074248410373750

PMID:20601591

Abstract

BACKGROUND

Gap junctions, which serve as intercellular channels that allow the passage of ions and other small molecules between neighboring cells, play an important role in vital functions, including the regulation of cell growth, differentiation, and development. Statins, the 3-hydroxy-3-methylglutaryl-coenzymeA (HMG-CoA) reductase inhibitors, have been shown to inhibit the migration and proliferation of smooth muscle cells (SMCs) leading to an antiproliferative effect. Recent studies have shown that statins can reduce gap junction protein connexin43 (Cx43) expression both in vivo and in vitro. However, little work has been done on the effects of statins on gap junctional intercellular communication (GJIC). We hypothesized in this study that lovastatin inhibits vascular smooth muscle cells (VSMCs) migration through the inhibition of the GJIC.

METHODS

Rat aortic SMCs (RASMCs) were exposed to lovastatin. Vascular smooth muscle cells migration was then assessed with a Transwell migration assay. Gap junctional intercellular communication was determined by using fluorescence recovery after photobleaching (FRAP) analysis, which was performed with a laser-scanning confocal microscope.

RESULTS

The migration of the cultured RASMCs were detected by Transwell system. Cell migration was dose-dependently inhibited with lovastatin. Compared with that in the control (110 ± 26), the number of migrated SMCs was significantly reduced to 72 ± 24 (P < .05), 62 ± 18 (P < .01), and 58 ± 19 (P < .01) at the concentration of 0.4, 2, and 10 umol/L, per field. The rate of fluorescence recovery (R) at 5 minutes after photobleaching was adopted as the functional index of GJIC. The R- value of cells exposed to lovastatin 10 umol/L for 48 hours was 24.38% ± 4.84%, whereas the cells in the control group had an R- value of 36.11% ± 10.53%, demonstrating that the GJIC of RASMCs was significantly inhibited by lovastatin (P < .01). Smaller concentrations of lovastatin 0.08 umol/L did not change gap junction coupling (P > .05).

CONCLUSIONS

These results suggest that lovastatin inhibits migration in a dose-dependent manner by attenuating JIC. Suppression of gap junction function could add another explanation of statin-induced antiproliferative effect.

摘要

背景

间隙连接作为细胞间的通道，允许离子和其他小分子在相邻细胞之间传递，在包括细胞生长、分化和发育在内的重要功能中发挥作用。他汀类药物，即 3-羟基-3-甲基戊二酰辅酶 A（HMG-CoA）还原酶抑制剂，已被证明可抑制平滑肌细胞（SMCs）的迁移和增殖，从而产生抗增殖作用。最近的研究表明，他汀类药物可在体内和体外降低间隙连接蛋白连接蛋白 43（Cx43）的表达。然而，关于他汀类药物对间隙连接细胞间通讯（GJIC）的影响，研究甚少。在本研究中，我们假设洛伐他汀通过抑制 GJIC 来抑制血管平滑肌细胞（VSMCs）的迁移。

方法

用洛伐他汀处理大鼠主动脉平滑肌细胞（RASMCs）。然后通过 Transwell 迁移测定法评估血管平滑肌细胞的迁移。通过荧光恢复后光漂白（FRAP）分析来确定间隙连接细胞间通讯，该分析使用激光扫描共聚焦显微镜进行。

结果

通过 Transwell 系统检测培养的 RASMCs 的迁移。洛伐他汀呈剂量依赖性地抑制细胞迁移。与对照组（110 ± 26）相比，浓度为 0.4、2 和 10 μmol/L 时，迁移的 SMC 数量分别显著减少至 72 ± 24（P <.05）、62 ± 18（P <.01）和 58 ± 19（P <.01）/视野。光漂白后 5 分钟时的荧光恢复率（R）被用作 GJIC 的功能指数。用洛伐他汀 10 μmol/L 处理 48 小时的细胞的 R 值为 24.38% ± 4.84%，而对照组细胞的 R 值为 36.11% ± 10.53%，表明洛伐他汀显著抑制了 RASMCs 的 GJIC（P <.01）。较小浓度的洛伐他汀 0.08 μmol/L 不会改变间隙连接偶联（P >.05）。