自主神经递质在乳鼠心房肌细胞Ca-L和KAch通道表达中的作用

发布时间：2018-06-28 04:13

本文选题：自主神经系统 + 离子通道　；参考：《遵义医学院》2017年硕士论文

【摘要】：目的:探讨不同浓度的自主神经递质去甲肾上腺素(NE)和乙酰胆碱(Ach))对乳鼠心房肌细胞L型钙离子通道(Ca-L)和乙酰胆碱敏感性钾通道(KAch)相关基因和蛋白表达的影响。方法:1天龄SD乳鼠,消化分离获得心房肌细胞,培养24h后干预分组。分为低药物浓度组、高药物浓度组。两药物浓度组分别按如下方案各自分成5亚组。(1)低浓度组:(1)对照组:不予干预,培养24h。(2)NE组:NE 0.1μmol/L干预24h。(3)Ach组:Ach 0.1μmol/L干预24h。(4)NE+Ach组:NE 0.1μmol/L+Ach0.1μmol/L干预24h;(5)NE+Meto(美托洛尔)组:NE 0.1μmol/L+Meto 0.1μmol/L干预24h。(2)高浓度组:(1)对照组:不予干预,培养72h。(2)NE组:NE 10μmol/L干预72h。(3)Ach组:Ach 10μmol/L干预72h。(4)NE+Ach组:NE 10μmol/L+Ach10μmol/L干预72h。(5)NE+Meto组:NE 10μmol/L+Meto 10μmol/L干预72h。实时荧光定量逆转录PCR、免疫印迹(WB)检测各组Cav1.2、Cav1.3、Kir3.1、Kir3.4 m RNA及蛋白表达水平。结果:(1)低浓度组:给予NE刺激24h后Cav1.2、Cav1.3 m RNA及蛋白表达上调,Kir3.1、Kir3.4 m RNA及蛋白表达下调;Ach刺激24h后Cav1.2、Cav1.3m RNA及蛋白表达下调,Kir3.1、Kir3.4 m RNA及蛋白表达上调;NE+Ach组与NE组比较,Cav1.2、Cav1.3 m RNA及蛋白表达下调,Kir3.1、Kir3.4 m RNA及蛋白表达上调;NE+Ach组与Ach组比较,Kir3.4 m RNA及蛋白表达下调;NE+Meto组与NE组比较,Meto干预后Cav1.2、Cav1.3 m RNA及蛋白表达下调,Kir3.1、Kir3.4 m RNA及蛋白表达上调。(2)高浓度组中:NE、Ach刺激72h后Cav1.2、Cav1.3、Kir3.1、Kir3.4 m RNA及蛋白均表达下调;NE+Ach组分别与NE组、Ach组比较,Kir3.1、Kir3.4蛋白表达均下调;NE+Meto组与NE组比较,Cav1.2、Cav1.3 m RNA及蛋白均表达上调,Kir3.1、Kir3.4蛋白表达下调。结论:(1)低浓度NE促进培养乳鼠心房肌细胞Cav1.2、Cav1.3基因和蛋白表达,抑制Kir3.1、Kir3.4表达;迷走神经则与之相反。Meto干预可抑制NE上述效应。(2)高浓度NE、Ach抑制Ca-Lh和KAch通道Cav1、2、Cav1.3及Kir3.1、Kir3.4 m RNA和蛋白表达。Meto干预可部分抑制上述效应。
[Abstract]:Aim: to investigate the effects of different concentrations of autonomic neurotransmitter norepinephrine (NE) and acetylcholine (Ach) on the expression of genes and proteins related to L-type calcium channel (Ca-L) and acetylcholine sensitive potassium channel (KAch) in neonatal rat atrial myocytes. Methods Atrial myocytes were isolated from 1 day old Sprague-Dawley rats and cultured for 24 hours. They were divided into low drug concentration group and high drug concentration group. The two drug concentration groups were divided into 5 subgroups according to the following scheme: (1) low concentration group: (1) control group: no intervention, (2) NE group: NE 0.1 渭 mol / L intervention 24 h. (3) Ach group: Ach 0.1 渭 mol / L intervention 24 h. (4) NE Ach group: NE 0.1 渭 mol / L Ach0.1 渭 mol / L for 24 h; (5) NE Meto 0.1 渭 mol / L Meto 0.1 渭 mol / L for 24 h. (2) High concentration group: (1) control group: no intervention. (2) NE group: 10 渭 mol / L 1: 10 渭 mol / L for 72 h; (3) Ach group: 10 渭 mol / L for 72 h; (4) NE group 10 渭 mol / L Ach10 渭 mol / L for 72 h; (5) NE group 10 渭 mol / L Meto 10 渭 mol / L for 72 h; (2) NE group 10 渭 mol / L Meto 10 渭 mol / L for 72 h; (2) NE group 10 渭 mol / L Ach10 渭 mol / L for 72 h; (5) NE group 10 渭 mol / L Meto 10 渭 mol / L for 72 h. Real-time fluorescence quantitative reverse transcription (PCR) and Western blotting (WB) were used to detect the expression of Cav1.2Cav1.3Cav1.3Cav1.1Kir3.4 mRNA and protein. Results: (1) in the low concentration group, the mRNA and protein expression of Cav1.2Cav1.3 mRNA and protein up-regulated Kir3.1 and Kir3.4 mRNA and protein expression after 24 hours of NE stimulation. After 24 hours of stimulation by Ach, the mRNA and protein expression of Cav1.2Cav1.3m and protein down-regulated Kir3.1Cav1.4mRNA and protein expression in NE Ach group were lower than those in NE group, compared with NE group, the expression of Cav1.2Cav1.3 mRNA in NE group was higher than that in NE group. The expression of Kir3.4 m RNA and protein in NE Ach group was lower than that in Ach group. The expression of Kir3.4 m RNA and protein in NE Meto group was significantly lower than that in NE group. (2) the expression of Kir3.1 ~ Kir3.4 m RNA and protein in NE Meto group was significantly higher than that in NE group. (2) the expression of Kir3.1 ~ Kir3.4 m mRNA and protein in NE Meto group was significantly higher than that in NE group. (2) the expression of Kir3.1 ~ Kir3.4 m mRNA and protein in NE Meto group was significantly higher than that in NE group. The expression of Cav1.2Cav1.3Cav1.3Cav3.1and Kir3.4m mRNA and protein were down-regulated in NE Ach group and NE group, respectively. The expression of Kir3.1 Kir3.4 protein was down-regulated in NE Meto group compared with NE Meto group and NE group. Both mRNA and protein expression of Cav1.2Cav1.3 m mRNA and protein were up-regulated and down-regulated in NE Ach group and NE group respectively. Conclusion: (1) low concentration of NE promotes the expression of Cav1.2Cav1.3 gene and protein in cultured rat atrial myocytes, and inhibits the expression of Kir3.1 and Kir3.4; (2) High concentration of NE-Ach inhibited Ca-Lh and KAch channel Cav1.3 and Kir3.1 Kir3.4 mRNA and protein expression. Meto partially inhibited these effects.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.75

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