Endothelial cell membrane cholesterol content regulates the contribution of TRPV4 channels in ACh-induced vasodilation in rat gracilis arteries

IF 1.9 4区医学 Q3 HEMATOLOGY Microcirculation Pub Date : 2022-06-11 DOI:10.1111/micc.12774

Emily E. Morin, Sophia Salbato, Benjimen R. Walker, Jay S. Naik

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引用次数: 1

Abstract

Objective

Our previous work demonstrated that endothelial cell (EC) membrane cholesterol is reduced following 48 h of chronic hypoxia (CH). CH couples endothelial transient receptor potential subfamily V member 4 (TRPV4) channels to muscarinic receptor signaling through an endothelium-dependent hyperpolarization (EDH) pathway does not present in control animals. TRVPV4 channel activity has been shown to be regulated by membrane cholesterol. Hence, we hypothesize that acute manipulation of endothelial cell membrane cholesterol inversely determines the contribution of TRPV4 channels to endothelium-dependent vasodilation.

Methods

Male Sprague–Dawley rats were exposed to ambient atmospheric (atm.) pressure or 48-h of hypoxia (0.5 atm). Vasodilation to acetylcholine (ACh) was determined using pressure myography in gracilis arteries. EC membrane cholesterol was depleted using methyl-β-cyclodextrin (MβCD) and supplemented with MβCD-cholesterol.

Results

Inhibiting TRPV4 did not affect ACh-induced vasodilation in normoxic controls. However, TRPV4 inhibition reduced resting diameter in control arteries suggesting basal activity. TRPV4 contributes to ACh-induced vasodilation in these arteries when EC membrane cholesterol is depleted. Inhibiting TRPV4 attenuated ACh-induced vasodilation in arteries from CH animals that exhibit lower EC membrane cholesterol than normoxic controls. EC cholesterol repletion in arteries from CH animals abolished the contribution of TRPV4 to ACh-induced vasodilation.

Conclusion

Endothelial cell membrane cholesterol impedes the contribution of TRPV4 channels in EDH-mediated dilation. These results provide additional evidence for the importance of plasma membrane cholesterol content in regulating intracellular signaling and vascular function.

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内皮细胞膜胆固醇含量调节TRPV4通道在乙酰胆碱诱导大鼠股薄动脉血管舒张中的作用

目的我们之前的研究表明，慢性缺氧(CH) 48小时后内皮细胞(EC)膜胆固醇降低。CH偶联内皮瞬时受体电位亚家族V成员4 (TRPV4)通道通过内皮依赖性超极化(EDH)途径传递毒毒碱受体信号，在对照动物中不存在。TRVPV4通道活性已被证明受膜胆固醇调节。因此，我们假设内皮细胞膜胆固醇的急性操纵与TRPV4通道对内皮依赖性血管舒张的贡献相反。方法雄性Sprague-Dawley大鼠暴露于环境大气压(atm)或缺氧48 h (0.5 atm)。用压力肌图测定股薄肌动脉对乙酰胆碱(ACh)的血管舒张。用甲基β-环糊精(m -β- cd)去除EC膜胆固醇，并添加m -β cd -胆固醇。结果抑制TRPV4对乙酰胆碱所致的血管舒张无影响。然而，TRPV4抑制降低了对照动脉的静息直径，提示基础活性。当EC膜胆固醇耗尽时，TRPV4有助于乙酰胆碱诱导的这些动脉血管舒张。抑制TRPV4可减弱乙酰胆碱诱导的动脉血管舒张，这些动物的EC膜胆固醇低于正常氧合对照组。CH动物动脉中EC胆固醇的补充消除了TRPV4对乙酰胆碱诱导的血管舒张的作用。结论内皮细胞膜胆固醇阻碍了TRPV4通道在edh介导的扩张中的作用。这些结果为质膜胆固醇含量在调节细胞内信号和血管功能中的重要性提供了额外的证据。

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来源期刊

Microcirculation 医学-外周血管病

CiteScore

5.00

自引率

4.20%

发文量

审稿时长

6-12 weeks

期刊介绍： The journal features original contributions that are the result of investigations contributing significant new information relating to the vascular and lymphatic microcirculation addressed at the intact animal, organ, cellular, or molecular level. Papers describe applications of the methods of physiology, biophysics, bioengineering, genetics, cell biology, biochemistry, and molecular biology to problems in microcirculation. Microcirculation also publishes state-of-the-art reviews that address frontier areas or new advances in technology in the fields of microcirculatory disease and function. Specific areas of interest include: Angiogenesis, growth and remodeling; Transport and exchange of gasses and solutes; Rheology and biorheology; Endothelial cell biology and metabolism; Interactions between endothelium, smooth muscle, parenchymal cells, leukocytes and platelets; Regulation of vasomotor tone; and Microvascular structures, imaging and morphometry. Papers also describe innovations in experimental techniques and instrumentation for studying all aspects of microcirculatory structure and function.