SK3表达在雌二醇介导的大鼠结肠平滑肌收缩中的作用
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国家自然科学基金青年基金(81400599)


The effect of small⁃conductance Ca2+⁃activated K+ channel 3 expression on relaxation of rat colonic smooth muscle regulated by estrogen
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    摘要:

    目的:探讨小电导钙激活钾通道3(SK3)表达在17β-雌二醇(E2)介导的大鼠结肠平滑肌收缩中的作用及机制。方法:将24只雄性SD大鼠随机分为4组。30 mm硅胶管皮下植入大鼠背部,内含不同溶液。分为对照组(C),内含溶剂;生理剂量组(EP)内含E2 0.3 mg/mL,使大鼠血清雌激素在生理浓度;超剂量组(ES)内含E2 0.9 mg/mL,使大鼠血清雌激素超生理浓度;ER抑制剂+E2组(EI)内含相同摩尔浓度E2和雌激素受体抑制剂(ICI 182780)。各组干预14 d后处理,免疫荧光及Western blot检测结肠平滑肌组织SK3分布及表达;MPA分析系统记录肌条收缩张力及对卡巴胆碱(Cch)刺激反应。E2孵育结肠平滑肌细胞(SMC)24 h后及过表达SK3后,激光共聚焦显微镜下动态观察Cch刺激后SMC内Ca2+变化。结果:Cch刺激后,EP组及ES组肌条收缩明显低于其他各组(P < 0.05)。 EP组及ES组平滑肌组织SK3表达高于C组及EI组,其中ES组有统计学意义(P < 0.05)。Cch刺激后,E2孵育组及过表达SK3组胞内Ca2+上升幅度较相应对照组显著降低(P < 0.05)。结论:E2可能通过促进SK3表达,减少Ca2+内流从而抑制结肠收缩。

    Abstract:

    Objective:The aim of the study was to explore the effect and possible mechanism of 17 β-estradiol(E2)on the expression of small conductance Ca2+ activated K+ channel 3(SK3)in rat colonic smooth muscle. Method:24 male Sprague-Dawley(SD)rats were randomly divided into 4 groups. Silicone tubes containing different solution was subcutaneously implanted in different groups of rats. Group C was filled with the solvent;Group EP was E2 in corn oil(0.3 mg/mL)which can keep serum levels of E2 at physiological levels;Group ES was E2 in corn oil(0.9 mg/mL)which can keep serum levels of E2 at supraphysiologic levels;Group EI was ICI 182780(estrogen receptor antagonist)plus E2(0.3 mg/mL). The contraction of muscle strips and SK3 expression in colonic smooth muscles were detected at 14 days after tube implanted. SK3 were overexpressed in SMCs and Ca2+ concentration in cells were detected by laser scanning confocal microscope. Results:The contraction of muscle strips of EP group and ES group is weaker than other groups(P < 0.05)after Cch stimulating. And the SK3 expression was upregulated in EP and ES groups(P < 0.05). Ca2+ decreased in SMCs after Cch stimulating in SK3 overexpression group and E2 stimulating group compared with control group. Conclusion:These findings indicate that E2 may suppress contraction of colonic smooth muscle by upregulating expression of SK3 and inhibiting Ca2+ influx in rats.

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熊文婕,姜 亚,汤玉蓉,郑永娉,俞 汀,姜 玲,沈小雪,林 琳. SK3表达在雌二醇介导的大鼠结肠平滑肌收缩中的作用[J].南京医科大学学报(自然科学版),2018,(6):753-757,815

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  • 收稿日期:2017-07-19
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  • 在线发布日期: 2018-06-22
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