IL-1β通过过氧化氢双相调节大鼠主动脉平滑肌细胞大电导钙敏感钾通道活性研究

髙渊; 梁潇; 李红兵; 肖懿慧

doi:10.13191/j.chj.2018.0151

IL-1β通过过氧化氢双相调节大鼠主动脉平滑肌细胞大电导钙敏感钾通道活性研究

The study of IL-1β biphasic effect on large conductance calcium sensitive potassium channel activity in rat aortic smooth muscle cells through hydrogen peroxide

摘要

摘要: 目的观察不同时间点白介素（IL）-1β对大鼠主动脉平滑肌细胞（ASMCs）内过氧化氢（hydrogen peroxide，H₂O₂）浓度及大电导钙敏感钾通道（large conductance Ca²⁺-activated K⁺channel，BKCa）活性影响，探讨IL-1β时间依赖双相调节BKCa通道活性机制。方法 ①分组：生理盐水对照组（saline组）、IL-1β处理30 min组、过氧化氢酶（catalase）+IL-1β处理30 min组、IL-1β处理48 h组、catalase+IL-1β处理48 h组；②检测IL-1β处理大鼠ASMCs不同时间点细胞内H₂O₂水平变化；③运用膜片钳技术，观察各实验组BKCa通道单通道电流开放概率（NPo）变化。结果 ①与saline组（36.7±7.23）μmol/ml的结果相比较，IL-1β与ASMCs共培养30 min组H₂O₂的水平增至（79.1±8.93）μmol/ml （P<0.01）；共培养48 h组，H₂O₂水平明显上调至（116.4±12.77）μmol/ml （P<0.01）；H₂O₂清除剂catalase可完全逆转IL-1β对H₂O₂影响。②与saline组比较，IL-1β单独处理ASMCs 30 min组NPo明显增加（P<0.05）；加入catalase预处理细胞，与saline组比较NPo无明显变化，与IL-1β单独处理ASMCs 30 min组比较NPo减少（P<0.05）。IL-1β单独处理ASMCs 48 h组，与saline组比较，NPo明显减少（P<0.05）；加入catalase预处理细胞，与saline组比较NPo减少（P<0.05），同时与IL-1β单独处理ASMCs 48 h组比较NPo增加有统计学意义（P<0.05）。结论 IL-1β短时间处理ASMCs上调BKCa通道活性，低浓度H₂O₂介导这一过程；长时间处理则下调BKCa通道活性，其机制部分通过高浓度H₂O₂实现；BKCa通道可能是防治冠脉痉挛有前景的药物作用靶点。

Abstract: AIM To observe the effect of IL-1β on hydrogen peroxide (H₂O₂) concentration and large conductance Ca²⁺-activated K⁺ channel(BKCa) activity in rat aortic smooth muscle cells (ASMCs) at different time points and to explore the mechanism of IL-1β time dependent biphasic modulation of the activity of BKCa channels. METHODS Five groups included:normal saline control group (Group saline), IL-1β treatment 30 min group, catalase (H₂O₂ scavenger)+IL-1β treatment 30 min group, IL-1β treatment 48 h group, and catalase+IL-1β treatment 48 h group. Changes were observed of intracellular H₂O₂ levels in the ASMCs treated with IL-1β at different time points. A patch clamp technique was used to observe the change of BKCa single channel current open probability (NPo) in each experimental group. RESULTS Compared with the results of the saline group (36.7±7.23) μmol/ml, the level of H₂O₂ in the 30 min group co-cultured with IL-1β increased to (79.1±8.93) μmol/ml (P<0.01), and in the 48 h group co-cultured with IL-1β, the H₂O₂ level was up to (116.4±12.77) μmol/ml (P<0.01). H₂O₂ scavenger catalase can completely reverse the influence of IL-1β at either time piont. The IL-1β alone treated group with ASMCs for 30 min increased NPo significantly (P<0.05). When catalase pretreated with cells, no significant changes in NPo was observed when compared with the saline group, but decreased when compared with IL-1β alone treaed with ASMCs for 30 min (P<0.05). IL-1β alone treated with ASMCs for 48 h decreased NPo significantly (P<0.05). When catalase pretreated with cells, NPo increased when compared with saline group (P<0.05), but decreased when compared with IL-1β alone treaed with ASMCs for 48 h (P<0.05). CONCLUSION IL-1β short time treated with ASMCs increased the activity of BKCa channel, while low concentration H₂O₂ mediated the process. Long time treatment reduced the activity of BKCa channel, and its mechanism was partly achieved by high concentration H₂O₂; The BKCa channel may be a promising drug target for the prevention and treatment of coronary spasm.

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