Krjukova J, Osna N, Pilmane M
Latvian Postgraduate and Continuing Medical Education Institute, Riga.
Scand J Clin Lab Invest. 2000 Aug;60(5):419-27. doi: 10.1080/003655100750019332.
Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.
通过流式细胞术,对健康供体的静息和PHA(植物血凝素)激活的人外周血淋巴细胞(HPL)中不同类型钾通道的表达进行了评估。在静息外周血淋巴细胞中,应用钾离子通道阻断剂(一种电压依赖性钾通道(K(V))的选择性阻断剂)会导致淋巴细胞膜电位明显去极化,在内质网钙释放剂毒胡萝卜素存在的情况下,这种去极化会进一步增强。在活化的HPL中,利用细胞周期分析数据评估了各种类型钾通道的表达。与静息细胞相比,在G0/G1期PHA激活的淋巴细胞中,钾离子通道阻断剂诱导的膜电位去极化不会因毒胡萝卜素而增强,而在S期和G2/M期PHA激活的淋巴细胞中,我们能够观察到钾离子通道阻断剂诱导的膜电位去极化后膜电位的复极化。用蝎毒素(一种同时阻断K(V)和K(Ca)通道的非选择性药物)替代钾离子通道阻断剂可消除PHA激活的淋巴细胞中的上述效应。因此,K(V)通道在静息和活化的HPL中均有活性,并且在PHA诱导外周淋巴细胞激活后,K(Ca)通道的表达随细胞周期进展而出现。