Oguri Gaku, Nakajima Toshiaki, Kikuchi Hironobu, Obi Shotaro, Nakamura Fumitaka, Komuro Issei
Department of Cardiovascular Medicine, University of Tokyo, Tokyo 113-8655, Japan.
Department of Cardiovascular Medicine, Dokkyo Medical University, Tochigi, 321-0293, Japan.
Heliyon. 2020 Dec 31;7(1):e05816. doi: 10.1016/j.heliyon.2020.e05816. eCollection 2021 Jan.
The effects of allyl isothiocyanate (AITC), transient receptor potential ankyrin 1 (TRPA1) agonist, on cultured human cardiac fibroblasts were examined by measuring intracellular Ca concentration [Ca] and whole-cell voltage clamp techniques. AITC (200 μM) increased Ca entry in the presence of [Ca]. Ruthenium red (RR) (30 μM), and La (0.5 mM), a general cation channel blocker, inhibited AITC-induced Ca entry. Under the patch pipette filled with Cs- and EGTA-solution, AITC induced the current of a reversal potential (Er) of approximately +0 mV. When extracellular Na ion was changed by NMDG, the inward current activated by AITC was markedly reduced. La and RR inhibited the AITC-induced current. The conventional RT-PCR analysis, Western blot, and immunocytochemical studies showed TRPA1 mRNA and protein expression. The present study shows the first evidence for functional Ca-permeable nonselective cation currents induced by AITC, possibly via TRPA1 in human cardiac fibroblast.
通过测量细胞内钙浓度[Ca]和全细胞膜片钳技术，研究了异硫氰酸烯丙酯（AITC）（一种瞬时受体电位锚蛋白1（TRPA1）激动剂）对培养的人心脏成纤维细胞的影响。在存在[Ca]的情况下，AITC（200μM）增加了钙内流。钌红（RR）（30μM）和镧（0.5 mM）（一种通用阳离子通道阻滞剂）抑制了AITC诱导的钙内流。在充满铯和乙二醇双四乙酸（EGTA）溶液的膜片钳电极下，AITC诱导出反转电位（Er）约为+0 mV的电流。当细胞外钠离子被N-甲基-D-葡萄糖胺（NMDG）置换时，AITC激活的内向电流明显降低。镧和RR抑制了AITC诱导的电流。传统的逆转录聚合酶链反应（RT-PCR）分析、蛋白质印迹法和免疫细胞化学研究显示了TRPA1信使核糖核酸（mRNA）和蛋白表达。本研究首次证明了AITC可能通过TRPA1在人心脏成纤维细胞中诱导功能性钙通透非选择性阳离子电流。
