摘要
目的 观察二十二碳六烯酸（DHA）对视网膜动脉平滑肌细胞（RASMCs）大电导钙激活钾离子（BK）通道的激活作用。 方法 传代培养RASMCs，取第6～8代细胞用于膜片钳实验。采用膜内向外型单通道膜片钳实验技术，分别记录DHA浓度0.0、1.0、3.0、5.0、7.5、10.0 μmol/L作用下RASMCs的BK通道开放概率（NP0）。以0.0、1.0、5.0 μmol/L不同浓度的DHA干预RASMCs，并以此分为对照组、低剂量组及高剂量组。采用蛋白免疫印迹法（Western blot）检测RAMSCs的BK通道中β1亚基的相对蛋白表达量。 结果 DHA浓度为0.0、1.0、3.0、5.0、7.5、10.0 μmol/L时，RASMCs的BK通道NP0分别为0.044 4±0.001 2、0.081 2±0.004 2、0.209 0±0.006 1、0.310 5±0.005 3、0.465 0±0.007 8、0.497 7±0.014 5。随DHA浓度的升高，BK通道NP0增加。不同DHA浓度作用下RASMCs的BK通道NP0比较，差异有统计学意义（F=2.621，P＜0.05）。Western blot检测结果显示，对照组、低剂量组、高剂量组RASMCs的BK通道中β1亚基的相对蛋白表达量比较，差异无统计学意义（F=11.657，P＞0.05）。 结论 DHA可直接激活RASMCs的BK通道。
Objective To investigate the effects of docosahexenoic acid （DHA） on large conductance Ca2＋-activated K＋ （BK） channels in normal retinal artery smooth muscle cells （RASMCs）. Methods Cultured human RASMCs （6 th-8 th generations） were used to patch clamp experiment. The open probabihties （NP0） in BK channels with different concentrations （0.0, 1.0, 3.0, 5.0, 7.5, 10.0 μmol/L） of DHA were recorded by patch clamp technique in single channel configuration. RASMCs were intervened by different concentrations （0.0, 1.0, 5.0 μmol/L） of DHA as control group, low and high doses of DHA groups, respectively. The protein expressions of β subunit of BK channels in RASMCs from three groups were measured by Western blot. Results The NP0 of BK channels were 0.044 4±0.001 2, 0.081 2±0.004 2, 0.209 0±0.006 1, 0.310 5±0.005 3, 0.465 0±0.007 8 and 0.497 7±0.014 5 with perfusate of 0.0, 1.0, 3.0, 5.0, 7.5, 10.0 μmol/L DHA. DHA activated BK channels in a dose-dependent manner （F=2.621,P〈0.05）. There was no significant difference in the protein expression of control group, low and high doses of DHA groups （F=11.657,P〉0.05）. Conclusion DHA can directly activate BK channels, no increasing in subunit expression of BK channels.
出处
《中华眼底病杂志》
CSCD
北大核心
2017年第3期295-297,共3页
Chinese Journal of Ocular Fundus Diseases
基金
江苏省卫生厅面上项目（H201449）
无锡市卫生局青年项目（Q201405）
关键词
二十二碳六烯酸类
视网膜动脉
肌细胞
平滑肌
大电导钙激活钾通道
膜片钳术
Docosahexaenoic acids
Retinal artery
Myocytes, smooth muscle
Large-conductance calcium-activated potassium channels
Patch-clamp techniques
