Zuoying Armed Forces General Hospital, Kaohsiung, Taiwan.
Hum Exp Toxicol. 2009 Oct;28(10):655-63. doi: 10.1177/0960327109106972. Epub 2009 Sep 21.
The effect of 2,4,6-trimethyl-N-(meta-3-trifluoromethyl-phenyl)-benzenesulfonamide (m-3M3FBS), a presumed phospholipase C (PLC) activator, on cytosolic free Ca(2+) concentrations (Ca( 2+)) in Madin-Darby canine kidney (MDCK) cells is unclear. This study explored whether m-3M3FBS changed basal Ca(2+) levels in suspended MDCK cells using fura-2 as a Ca(2+)-sensitive fluorescent dye. M-3M3FBS at concentrations between 0.1 and 20 microM increased Ca(2+) in a concentration-dependent manner. The Ca(2+) signal was decreased by removing extracellular Ca(2+). M-3M3FBS-induced Ca(2+) influx was inhibited by the store-operated Ca(2+) channel blockers nifedipine, econazole, and SK&F96365, and by the phospholipase A2 inhibitor aristolochic acid. In Ca(2+)-free medium, 20-microM m-3M3FBS pretreatment abolished the Ca(2+) rise induced by the endoplasmic reticulum Ca(2+) pump inhibitors thapsigargin (TG) and cyclopiazonic acid (CPA). Conversely, pretreatment with TG or CPA partly reduced m-3M3FBS-induced Ca(2+) rise. The inhibition of PLC with U73122 did not alter m-3M3FBS-induced Ca(2+) rise. Collectively, in MDCK cells, m-3M3FBS induced Ca(2+) rises by causing PLC-independent Ca(2+) release from the endoplasmic reticulum and Ca(2+) influx via store-operated Ca(2+) channels and other unidentified Ca(2+) channels.
2,4,6-三甲基-N-(间-3-三氟甲基-苯基)苯磺酰胺(m-3M3FBS)被认为是一种磷脂酶 C(PLC)激活剂,其对 Madin-Darby 犬肾(MDCK)细胞胞浆游离 Ca(2+)浓度(Ca(2+))的影响尚不清楚。本研究使用荧光染料 fura-2 来探讨 m-3M3FBS 是否改变悬浮培养的 MDCK 细胞的基础 Ca(2+)水平。结果发现,浓度在 0.1 至 20μM 之间的 m-3M3FBS 可浓度依赖性地增加 Ca(2+)。去除细胞外 Ca(2+)可使 Ca(2+)信号减弱。m-3M3FBS 诱导的 Ca(2+)内流可被储存操纵的 Ca(2+)通道阻滞剂硝苯地平、依康唑和 SK&F96365 以及磷脂酶 A2 抑制剂马兜铃酸所抑制。在无 Ca(2+)的培养基中,20μM m-3M3FBS 预处理可消除内质网 Ca(2+)泵抑制剂 thapsigargin(TG)和环匹阿尼酸(CPA)引起的 Ca(2+)升高。相反,TG 或 CPA 的预处理部分减少了 m-3M3FBS 诱导的 Ca(2+)升高。U73122 抑制 PLC 不会改变 m-3M3FBS 诱导的 Ca(2+)升高。综上,在 MDCK 细胞中,m-3M3FBS 通过引起 PLC 非依赖性内质网 Ca(2+)释放以及通过储存操纵的 Ca(2+)通道和其他未识别的 Ca(2+)通道的 Ca(2+)内流引起 Ca(2+)升高。