内源性雌激素调控脓毒症小鼠心肌L型钙离子通道α1C亚基的表达

Endogenous estrogen regulates the expression of myocardial L-type calcium channel α1C subunit(CACNA1C) in septic mice

目的探讨内源性雌激素对脓毒症小鼠心肌钙离子通道α1C亚基(CACNA1C)的影响。方法雌性昆明小鼠行双侧卵巢摘除术,术后30 d利用盲肠结扎穿刺术(CLP)建立脓毒症小鼠模型,分为5组:脓毒症组(S组),去卵巢+脓毒症组(QS组),假手术组(C组),去卵巢+假手术组(QC组),去卵巢+苯甲酸雌二醇(QE组)。荧光定量PCR和Western blot检测造模后3 h,6 h,12 h,24 h左心室CACNA1C表达水平,免疫组化检测造模后12 h左心室CACNA1C表达及分布情况。结果(1)QC组CACNA1C的表达水平显著高于C组,QE组CACNA1C的表达水平显著低于QC组。(2)CLP后3 h,S组CACNA1C表达显著高于对照组和其它时间点。(3)CLP后3 h、6 h、12 h和24 h,QS组CACNA1C表达均显著低于对照组,且各时间点之间无差异。结论(1)双侧卵巢摘除后CACNA1C表达显著升高,双侧卵巢摘除后补给苯甲酸雌二醇可以逆转CACNA1C的升高。(2)正常小鼠CLP后3 h CACNA1C的表达出现短暂的升高,而双侧卵巢摘除的小鼠CLP后各时间点CACNA1C的表达均显著低于对照组。

Objective To investigate the effect of endogenous estrogen on myocardial L-type calcium channelα1C subunit(CACNA1C)in septic mice.Methods Female Kunming mice underwent bilateral ovarian ablation,a model of sepsis was established by cecal ligation puncture(CLP)30 days after surgery.The experimental mice were divided into 5 groups:sepsis group(S group),ovariectomy+sepsis group(QS group),sham group(C group),ovariectomy+sham group(QC group),ovariectomy+estradiol benzoate(QE group).The expression levels of left ventricular CACNA1C were detected by real-time PCR and Western blot at 3 h,6 h,12 h,and 24 h after CLP.The expression and distribution of CACNA1C was detected by immunohistochemistry 3 h after CLP.Results The expression of CACNA1C in QC group was significantly higher than that in the C group,while the expression level of CACNA1C in the QE group was significantly lower than that in the QC group.At 3 h after CLP,the expression of CACNA1C in the S group was significantly higher than that in the control and other time points.At 3 h,6 h,12 h and 24 h after CLP,the expression of CACNA1C in the QS group was significantly lower than that in the control group,and there was no significant difference among the four time points.Conclusion The expression of CACNA1C is significantly increased after bilateral ovarian ablation.Replenishment of estradiol benzoate after lateral ovarian ablation can reverse the increase of CACNA1C.The expression of CACNA1C is transiently increased at 3 h after CLP in normal mice,while the expression of CACNA1C at each time point after CLP in bilateral ovarian-extracted mice is significantly lower than that in the control group.