Cytokine effects on gap junction communication and connexin expression in human bladder smooth muscle cells and suburothelial myofibroblasts.
Cytokine effects on gap junction communication and connexin expression in human bladder smooth muscle cells and suburothelial myofibroblasts.
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DOI：
10.1371/journal.pone.0020792
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Neuhaus J
中科院分区：
文献类型：
--
作者：
Heinrich M;Oberbach A;Schlichting N;Stolzenburg JU;Neuhaus J
近十年来，细胞因子被认为是一组主要的局部细胞信号分子，与间质性膀胱炎(IC)和膀胱过度活动症(OAB)等膀胱功能障碍有关。缝隙连接细胞间通讯(GJIC)对正常膀胱功能的协调是必不可少的，已发现在膀胱功能障碍时会发生改变。间隙连接蛋白(Cx43)和Cx45是膀胱平滑肌细胞(HBSMC)和上皮下肌纤维母细胞(HsMF)中最重要的缝隙连接蛋白。细胞因子对缝隙连接蛋白表达的调节在多种组织中均有发现。因此，我们研究了白介素4、白介素6、白介素10、肿瘤坏死因子α和转化生长因子β1对人膀胱平滑肌细胞和人上皮下肌成纤维细胞GJIC和Cx43、Cx45表达的影响。从膀胱切除患者的膀胱组织中分离培养HBSMC和hsMF。在细胞因子刺激下，用荧光漂白后恢复法(FRAP)分析hBSMC和hsMF的GJIC。用定量聚合酶链式反应和共聚焦免疫荧光法检测Cx43和Cx45的表达。斑点印迹法定量Cx43和Cx45的膜蛋白组分。IL-6刺激后，两种细胞的细胞-细胞-通讯均上调。在hBSMC中，IL 4和转化生长因子β1均降低GJIC和Cx43的蛋白表达，而肿瘤坏死因子α不改变FRAP实验中的通讯，而增加Cx43的表达。GJ斑块大小与测得的偶联效率相关，而Cx45的表达与GJIC的调节无关。我们对GJIC的特异性细胞因子作用的发现支持了细胞因子在OAB和IC的病理生理学中起关键作用的观点。有趣的是，这些作用与促炎和抗炎细胞因子的经典定义无关。我们的结论是，连接蛋白的调节涉及基因组和/或翻译后事件，并且hBSMC和hsMF中的GJIC依赖于Cx43而不是Cx45。
The last decade identified cytokines as one group of major local cell signaling molecules related to bladder dysfunction like interstitial cystitis (IC) and overactive bladder syndrome (OAB). Gap junctional intercellular communication (GJIC) is essential for the coordination of normal bladder function and has been found to be altered in bladder dysfunction. Connexin (Cx) 43 and Cx45 are the most important gap junction proteins in bladder smooth muscle cells (hBSMC) and suburothelial myofibroblasts (hsMF). Modulation of connexin expression by cytokines has been demonstrated in various tissues. Therefore, we investigate the effect of interleukin (IL) 4, IL6, IL10, tumor necrosis factor-alpha (TNFα) and transforming growth factor-beta1 (TGFβ1) on GJIC, and Cx43 and Cx45 expression in cultured human bladder smooth muscle cells (hBSMC) and human suburothelial myofibroblasts (hsMF). HBSMC and hsMF cultures were set up from bladder tissue of patients undergoing cystectomy. In cytokine stimulated cultured hBSMC and hsMF GJIC was analyzed via Fluorescence Recovery after Photo-bleaching (FRAP). Cx43 and Cx45 expression was assessed by quantitative PCR and confocal immunofluorescence. Membrane protein fraction of Cx43 and Cx45 was quantified by Dot Blot. Upregulation of cell-cell-communication was found after IL6 stimulation in both cell types. In hBSMC IL4 and TGFβ1 decreased both, GJIC and Cx43 protein expression, while TNFα did not alter communication in FRAP-experiments but increased Cx43 expression. GJ plaques size correlated with coupling efficacy measured, while Cx45 expression did not correlate with modulation of GJIC. Our finding of specific cytokine effects on GJIC support the notion that cytokines play a pivotal role for pathophysiology of OAB and IC. Interestingly, the effects were independent from the classical definition of pro- and antiinflammatory cytokines. We conclude, that connexin regulation involves genomic and/or post-translational events, and that GJIC in hBSMC and hsMF depend of Cx43 rather than on Cx45.
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影响因子：
23.4
作者：
Kuijpers, Kamiel A. J.;Heesakkers, John P. F. A.;Schalken, Jack A.
通讯作者：
Schalken, Jack A.
影响因子：
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作者：
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