Thapsigargin-resistant thiacalix[4]arene C-1087-sensitive component of the contractile activity in rat myometrium reflects the functioning of plasma membrane calcium pump

Q4 Agricultural and Biological Sciences
Olga Tsymbalyuk, Tetyana Veklich, Roman Rodik, Sergiy Karakhim, Sergiy Vyshnevskyi, Vitaly Kalchenko, Sergiy Kosterin
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Abstract

Background. According to existing knowledge, thiacalix[4]arene С-1087 is highly capable of inhibiting Са2+-pump (Са2+, Mg2+-АТРase) of the plasma membrane; at the same time it inhibits the functioning of Са2+-pump of the sarcoplasmic reticulum of uterine smooth muscles to some degree. The aim of this research was to study the effects of С-1087 on the concentration of Ca2+ ions and contractile activity of the rat myometrium cells using an inhibitor of Са2+-pump of the sarcoplasmic reticulum – thapsigargin. Materials and Methods. The experiments were conducted using outbred white non-pregnant rats. The contractile activity in the preparations of longitudinal SM of uterine horns with preserved endothelium was registered in the isometric mode. To determine the changes in [Ca2+]i level, myocytes were treated with probes Hoechst 33342 (to test the nucleus of the cell) and fluo-4 AM (to test the change in Са2+-concentration in the cell). Results. The tenzometric studies with the subsequent mechanokinetic analysis demonstrated that under the action of thapsigargin (0.5 µM), thiacalix[4]arene C-1087 (10 µM) caused considerable changes in the kinetics of the spontaneous contractile activity processes in the myometrium of rats, including the decrease in the maximal contraction velocity and the increase in the maximal relaxation velocity. By means of confocal microscopy with Ca2+-sensitive fluorescent probe fluo-4, it was demonstrated that the application of thiacalix[4]arene С-1087 to immobilized myocytes of the uterus against the background of thapsigargin caused a transient spike of Са2+-signal with the subsequent turn of the intracellular concentration of Ca ions to the stable increased level. The effects of С-1087 under the action of thapsigargin regarding the relaxation phase in the spontaneous myometrium contractions were removed after the preliminary blocking of nitric oxide synthases L-NAME (100 µM). Under the action of L-NAME, thiacalix[4]arene С-1087 (10 µM) caused complete inhibition of the relaxation process in the contraction of myometrium preparations, induced by high-potassium solution (80 mM). Conclusions. The primary reason for changes in the contractile activity and Ca2+-signal in uterine myocytes under the effect of thiacalix[4]arene С-1087 is its ability to inhibit Са2+-pump of the plasma membrane; further С-1087-induced changes in the smooth muscle tissues may be caused by the increased level of Са2+ concentration in myocytes. The obtained results demonstrate thiacalix[4]arene С-1087 is a promising compound for the elaboration of pharmacological preparations for modulating the contractile activity in smooth muscles, including myometrium.
thapsigarin -resistant thiacalix[4]芳烃c -1087在大鼠肌层收缩活性中的敏感成分反映了质膜钙泵的功能
背景。根据现有的知识,噻吩[4]芳烃С-1087具有很强的抑制质膜Са2+-泵(Са2+, Mg2+-АТРase)的能力;同时在一定程度上抑制子宫平滑肌肌浆网Са2+-泵的功能。本研究的目的是研究С-1087对Ca2+离子浓度和大鼠肌膜细胞收缩活性的影响,使用Са2+-肌浆网泵抑制剂- thapsigargin。材料与方法。实验是用未怀孕的纯种大鼠进行的。用等长模型记录保存内皮的子宫角纵SM制剂的收缩活性。为了确定[Ca2+]i水平的变化,用探针Hoechst 33342(用于测试细胞核)和fluo-4 AM(用于测试细胞中Са2+-浓度的变化)处理肌细胞。结果。张力学研究和随后的力学动力学分析表明,在thapsigargin(0.5µM)的作用下,thiacalix[4]芳烃C-1087(10µM)引起大鼠肌层自发收缩活动过程动力学的显著变化,包括最大收缩速度降低和最大松弛速度增加。通过使用钙离子敏感荧光探针fluo-4共聚焦显微镜,我们发现在thapsigargin的背景下,将thiacalix[4]芳烯С-1087应用于固定的子宫肌细胞,会引起Са2+信号的短暂尖峰,随后细胞内钙离子浓度转向稳定的升高水平。初步阻断一氧化氮合酶L-NAME(100µM)后,可消除С-1087在thapsigargin作用下对自发性肌层收缩松弛期的影响。在L-NAME作用下,thiacalix[4]芳烃С-1087(10µM)完全抑制高钾溶液(80 mM)诱导的肌层制剂收缩的松弛过程。结论。thiacalix[4]芳烃С-1087作用下子宫肌细胞收缩活性和Ca2+信号发生变化的主要原因是其能够抑制质膜的Са2+泵;平滑肌组织的进一步С-1087-induced变化可能是由肌细胞中Са2+浓度水平的增加引起的。所获得的结果表明噻烷[4]芳烃С-1087是一种很有前途的化合物,可用于制备调节平滑肌(包括肌层)收缩活性的药理制剂。
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来源期刊
CiteScore
0.60
自引率
0.00%
发文量
22
审稿时长
5 weeks
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