三碘甲腺原氨酸对缺氧/复氧致乳小鼠心室肌细胞钙离子通道异常的影响及其信号通路机制

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摘要目的观察三碘甲腺原氨酸（T3）对缺氧/复氧（H/R）乳小鼠心室肌细胞钙离子通道异常的影响及PI3K/Akt信号通路在其中的作用。方法分离乳小鼠心室肌细胞并建立细胞H/R模型，设置正常对照（Control）组、H/R组、T3+H/R组、T3+H/R+LY294002（LY）组、H/R+LY组。利用膜片钳技术，观察L-型钙离子通道电流（ICaL）的变化，Western blot检测心肌细胞PI3K/Akt信号通路的表达。结果与Control组比较，H/R组ICaL峰值电流减小（P ＜ 0.01），I-V曲线上移；与H/R组比较，T3+H/R组ICaL峰值电流增大（P ＜ 0.01），H/R+LY组ICaL峰值电流减小（P ＜ 0.01）；与T3+H/R组比较，T3+H/R+LY组ICaL峰值电流减小（P ＜ 0.01）。与Control组比较，H/R组V1/2明显减小（P < 0.05），失活曲线右移，失活减慢。与H/R组比较，T3+H/R组V1/2增大（P < 0.05），失活曲线左移，失活加快。与Control组比较，H/R组V1/2升高（P < 0.05），激活曲线左移，激活加快；与H/R组比较，T3+H/R组V1/2减小（P < 0.05），激活曲线右移，激活减慢。与Control组比较，H/R组心肌细胞P-PI3K及P-Akt表达减少（P < 0.01）；与H/R组比较，T3+H/R组心肌细胞P-PI3K及P-Akt表达增加（P ＜ 0.01），H/R+LY组心肌细胞P-PI3K及P-Akt表达减少（P ＜ 0.01）；与T3+H/R组比较，T3+H/R+LY组心肌细胞P-PI3K及P-Akt表达减少（P ＜ 0.01）。结论心肌细胞H/R时ICaL电流减小，影响心肌细胞兴奋收缩，而T3可以通过激活PI3K/Akt信号通路保护L-型钙离子通道，减少通道电流的下降，从而发挥其心脏保护作用。

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	刘磊曾彬廖小婷

关键词 ：甲状腺激素, 心室肌细胞, 缺氧/复氧, L-型钙离子通道

Abstract：Objective To observe the effect of triiodothyronine (T3) on abnormal calcium channels in ventricular myocytes of hypoxia/reoxygenation (H/R) lactating mice and the role of PI3K/Akt signaling pathway in it. Methods Isolation of ventricular myocytes from neonatal mice and establishment of cell H/R model were carried out. Normal control (Control) group, H/R group, T3+H/R group, T3+H/R+LY294002 (LY) group and H/R+LY group were set up. Patch clamp technique was used to observe the changes of L-type calcium channel current (ICaL) and Western blot was used to detect the expression of PI3K/Akt signaling pathway in cardiomyocytes. Results Compared with control group, the peak current of ICaL in H/R group decreased (P < 0.01) and I-V curve moved up. Compared with H/R group, ICaL peak current increased in T3+H/R group (P < 0.01), and decreased in H/R+LY group (P < 0.01). Compared with T3+H/R group, ICaL peak current decreased in T3+H/R+LY group (P < 0.01). Compared with control group, V1/2 in H/R group decreased significantly (P < 0.05), inactivation curve shifted to the right and inactivation slowed down. Compared with H/R group, V1/2 of T3+H/R group increased (P < 0.05), the inactivation curve moved left, and the inactivation accelerated. Compared with control group, V1/2 of H/R group increased (P < 0.05), the activation curve moved left, and the activation accelerated. Compared with H/R group, V1/2 of T3+H/R group decreased (P < 0.05), the activation curve moved right and the activation slowed down. Compared with control group, the expression of P-PI3K and P-Akt decreased in H/R group (P < 0.01). Compared with H/R group, the expression of P-PI3K and P-Akt in myocardial cells of T3+H/R group increased (P < 0.01), while the expression of P-PI3K and P-Akt in myocardial cells of H/R+LY group decreased (P < 0.01). Compared with T3+H/R group, the expression of P-PI3K and P-Akt in myocardial cells of T3+H/R+LY group decreased (P < 0.01). Conclusion ICaL current decreases when cardiomyocyte H/R, affecting excitation and contraction of cardiomyocyte. T3 can protect L-type calcium channel by activating PI3K/Akt signaling pathway and reduce the decrease of channel current, thus exerting its cardioprotective effect.

Key words： Thyroid hormone Ventricular cardiomyocytes Hypoxia/reoxygenation L-type calcium channel

基金资助:国家自然科学基金资助项目（81570333）。

通讯作者: 曾彬（1978.11-），男，博士，主任医师，副教授；研究方向：冠心病基础与临床。

作者简介: 刘磊（1992.4-），女，硕士；研究方向：冠心病基础与临床。

引用本文:

刘磊曾彬廖小婷. 三碘甲腺原氨酸对缺氧/复氧致乳小鼠心室肌细胞钙离子通道异常的影响及其信号通路机制[J]. 中国医药导报, 2019, 16(29): 11-15.
LIU Lei ZENG Bin LIAO Xiaoting. Effect of triiodothyronine on abnormal calcium channel in ventricular myocytes of hypoxia/reoxygenation-induced lactating mice and its signaling pathway mechanism. 中国医药导报, 2019, 16(29): 11-15.

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http://www.yiyaodaobao.com.cn/CN/ 或 http://www.yiyaodaobao.com.cn/CN/Y2019/V16/I29/11

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