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首页> 外文期刊>Journal of Cellular Physiology >High glucose alters apoptosis and proliferation in HEK293 cells by inhibition of cloned BK Ca channel.
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High glucose alters apoptosis and proliferation in HEK293 cells by inhibition of cloned BK Ca channel.

机译:高葡萄糖改变细胞凋亡和增殖HEK293细胞克隆BK Ca的抑制通道。

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It has been reported that diabetic vascular dysfunction is associated with impaired function of large conductance Ca(2+) -activated K(+) (BK(Ca) ) channels. However, it is unclear whether impaired BK(Ca) channel directly participates in regulating diabetic vascular remodeling by altering cell growth in response to hyperglycemia. In the present study, we investigated the specific role of BK(Ca) channel in controlling apoptosis and proliferation under high glucose concentration (25 mM). The cDNA encoding the alpha+beta1 subunit of BK(Ca) channel, hSloalpha+beta1, was transiently transfected into human embryonic kidney 293 (HEK293) cells. Cloned BK(Ca) currents were recorded by both whole-cell and cell-attached patch clamp techniques. Cell apoptosis was assessed with immunocytochemistry and analysis of fragmented DNA by agarose gel electrophoresis. Cell proliferation was investigated by flow cytometry assays, MTT test, and immunocytochemistry. In addition, the expression of anti-apoptotic protein Bcl-2, intracellular Ca(2+) , and mitochondrial membrane potential (Deltapsim) were also examined to investigate the possible mechanisms. Our results indicate that inhibition of cloned BK(Ca) channels might be responsible for hyperglycemia-altered apoptosis and proliferation in HEK-hSloalpha+beta1 cells. However, activation of BK(Ca) channel by NS1619 or Tamoxifen significantly induced apoptosis and suppressed proliferation in HEK-hSloalpha+beta1 cells under hyperglycemia condition. When rat cerebral smooth muscle cells were cultured in hyperglycemia, similar findings were observed. Moreover, the possible mechanisms underlying the activation of BK(Ca) channel were associated with decreased expression of Bcl-2, elevation of intracellular Ca(2+) , and a concomitant depolarization of Deltapsim in HEK-hSloalpha+beta1 cells. In conclusion, cloned BK(Ca) channel directly regulated apoptosis and proliferation of HEK293 cell under hyperglycemia condition.
机译:据报道,糖尿病血管功能与功能受损有关大电导钙激活(2 +)K (+)(BK (Ca))通道。是否受损BK (Ca)直接渠道参与调节糖尿病血管重构通过改变细胞生长反应高血糖。调查了BK通道(Ca)的特定角色在控制细胞凋亡和增殖高葡萄糖浓度(25毫米)。编码α+ beta1 BK (Ca)的亚基频道,hSloalpha + beta1是暂时性的转染到人类胚胎肾293(HEK293)细胞。全细胞和cell-attached记录膜片钳技术。用免疫细胞化学和分析评估支离破碎的DNA琼脂糖凝胶电泳。细胞增殖是调查流程血细胞计数化验、MTT测试和免疫细胞化学。抗凋亡蛋白bcl - 2,细胞内Ca(2 +),线粒体膜电位(Deltapsim)也进行调查可能的机制。抑制克隆BK (Ca)渠道负责hyperglycemia-altered细胞凋亡并在HEK-hSloalpha + beta1细胞增殖。然而,激活NS1619 BK通道(Ca)的或三苯氧胺显著诱导细胞凋亡在HEK-hSloalpha + beta1抑制增殖在高血糖条件下细胞。脑血管平滑肌细胞培养高血糖,类似的研究结果。此外,可能的机制BK (Ca)通道的激活有关bcl - 2的表达减少,海拔细胞内钙(2 +),和伴随的去极化的DeltapsimHEK-hSloalpha + beta1细胞。汉堡王(Ca)直接调控细胞凋亡和频道HEK293细胞扩散在高血糖条件。

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