Bae Hyemi, Kim Taeho, Lim Inja
Department of Physiology, College of Medicine, Chung-Ang University, Seoul 06974, Korea.
Department of Internal Medicine, College of Medicine, Chung-Ang University Hospital, Seoul 06973, Korea.
Korean J Physiol Pharmacol. 2022 Jan 1;26(1):25-36. doi: 10.4196/kjpp.2022.26.1.25.
To identify the effect and mechanism of carbon monoxide (CO) on delayed rectifier K currents () of human cardiac fibroblasts (HCFs), we used the wholecell mode patch-clamp technique. Application of CO delivered by carbon monoxidereleasing molecule-3 (CORM3) increased the amplitude of outward K currents, and diphenyl phosphine oxide-1 (a specific blocker) inhibited the currents. CORM3- induced augmentation was blocked by pretreatment with nitric oxide synthase blockers (L-NG-monomethyl arginine citrate and L-NG-nitro arginine methyl ester). Pretreatment with KT5823 (a protein kinas G blocker), 1H-[1,-2,-4] oxadiazolo-[4,-3-a] quinoxalin-1-on (ODQ, a soluble guanylate cyclase blocker), KT5720 (a protein kinase A blocker), and SQ22536 (an adenylate cyclase blocker) blocked the CORM3 stimulating effect on . In addition, pretreatment with SB239063 (a p38 mitogen-activated protein kinase [MAPK] blocker) and PD98059 (a p44/42 MAPK blocker) also blocked the CORM3's effect on the currents. When testing the involvement of -nitrosylation, pretreatment of N-ethylmaleimide (a thiol-alkylating reagent) blocked CO-induced activation and DL-dithiothreitol (a reducing agent) reversed this effect. Pretreatment with 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)-21H,23H porphyrin manganese (III) pentachloride and manganese (III) tetrakis (4-benzoic acid) porphyrin chloride (superoxide dismutase mimetics), diphenyleneiodonium chloride (an NADPH oxidase blocker), or allopurinol (a xanthine oxidase blocker) also inhibited CO-induced activation. These results suggest that CO enhances in HCFs through the nitric oxide, phosphorylation by protein kinase G, protein kinase A, and MAPK, -nitrosylation and reduction/oxidation (redox) signaling pathways.
为了确定一氧化碳（CO）对人心脏成纤维细胞（HCFs）延迟整流钾电流（）的影响及机制，我们采用了全细胞模式膜片钳技术。应用一氧化碳释放分子-3（CORM3）递送的CO可增加外向钾电流的幅度，而二苯基氧化膦-1（一种特异性阻滞剂）可抑制该电流。一氧化氮合酶阻滞剂（L-NG-单甲基精氨酸柠檬酸盐和L-NG-硝基精氨酸甲酯）预处理可阻断CORM3诱导的增强作用。用KT5823（一种蛋白激酶G阻滞剂）、1H-[1,2,4]恶二唑并-[4,3-a]喹喔啉-1-酮（ODQ，一种可溶性鸟苷酸环化酶阻滞剂）、KT5720（一种蛋白激酶A阻滞剂）和SQ22536（一种腺苷酸环化酶阻滞剂）预处理可阻断CORM3对的刺激作用。此外，用SB239063（一种p38丝裂原活化蛋白激酶[MAPK]阻滞剂）和PD98059（一种p44/42 MAPK阻滞剂）预处理也可阻断CORM3对电流的作用。在检测亚硝基化的参与情况时，N-乙基马来酰亚胺（一种硫醇烷基化试剂）预处理可阻断CO诱导的激活，而二硫苏糖醇（一种还原剂）可逆转这种作用。用5,10,15,20-四（1-甲基吡啶-4-基）-21H,23H卟啉锰（III）五氯化物和锰（III）四（4-苯甲酸）卟啉氯化物（超氧化物歧化酶模拟物）、二苯基碘鎓氯化物（一种NADPH氧化酶阻滞剂）或别嘌呤醇（一种黄嘌呤氧化酶阻滞剂）预处理也可抑制CO诱导的激活。这些结果表明，CO通过一氧化氮、蛋白激酶G、蛋白激酶A和MAPK的磷酸化、亚硝基化以及还原/氧化（氧化还原）信号通路增强HCFs中的。
