TGF-β1 Inhibits Cx43 Expression and Formation of Functional Syncytia in Cultured Smooth Muscle Cells from Human Detrusor
TGF-β1 Inhibits Cx43 Expression and Formation of Functional Syncytia in Cultured Smooth Muscle Cells from Human Detrusor
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DOI：
10.1016/j.eururo.2008.03.090
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发表时间：
2009-02-01
期刊：
影响因子：
23.4
通讯作者：
Stolzenburg, Jens-Uwe
中科院分区：
文献类型：
--
作者：
Neuhaus, Jochen;Heinrich, Marco;Stolzenburg, Jens-Uwe
背景：人逼尿肌平滑肌细胞 (hBSMC) 通过连接蛋白 43 (Cx43) 阳性间隙连接偶联形成功能性合胞体。间隙连接通讯可能对于同步逼尿肌收缩是必要的，并且应该在排尿障碍中改变。其他作者表明，多效性细胞因子 TGF-β 1 上调人主动脉平滑肌细胞中的 Cx43 表达。目的：在这项研究中，我们检查了 TGF-β 1 对培养的 hBSMC 中 Cx43 表达的影响。设计、设置和参与者：从接受膀胱切除术的患者中建立的 hBSMC 培养物用重组人 TGF-β 1 进行处理。测量：然后通过蛋白质印迹实时检查 Cx43 表达PCR 和免疫细胞化学。染料注射实验用于研究功能性合胞体的大小。结果和局限性：染料偶联实验揭示了传代细胞培养物（P1-P4）中功能性合胞体的稳定形成。 TGF-β1 刺激导致 Cx43 免疫反应性和偶联显着降低。 Cx43蛋白表达显着下调，Cx43 mRNA仅为对照水平的30%。有趣的是，Cx43 的低磷酸化种类尤其受到影响。结论：我们的实验表明，在培养的 hBSMC 中，TGF-β 1 对连接蛋白 43 具有显着下调作用。这些发现支持以下观点：TGF-β1 参与膀胱功能障碍的病理生理学。 (C) 2008 年欧洲泌尿外科协会。由 Elsevier B.V. 出版。保留所有权利。
Background: Human detrusor smooth muscle cells (hBSMCs) are coupled by connexin 43 (Cx43)-positive gap junctions to form functional syncytia. Gap junctional communication likely is necessary for synchronised detrusor contractions and is supposed to be altered in voiding disturbances. Other authors have shown that the pleiotropic cytokine TGF-beta 1 upregulates Cx43 expression in human aortic smooth muscle cells.Objective: In this study, we examined the TGF-beta 1 effects on Cx43 expression in cultured hBSMCs.Design, Setting, and Participants: hBSMC cultures, established from patients undergoing cystectomy, were treated with recombinant human TGF-beta 1. Measurements: Cx43 expression was then examined by Western blotting, real-time PCR, and immunocytochemistry. Dye-injection experiments were used to study the size of functional syncytia.Results and Limitations: Dye-coupling experiments revealed stable formation of functional syncytia in passaged cell cultures (P1-P4). Stimulation with TGF-beta 1 led to significant reduction of Cx43 immunoreactivity and coupling. Cx43 protein expression was significantly downregulated and Cx43 mRNA was only 30% of the control level. Interestingly, low phosphorylation species of Cx43 were particularly affected.Conclusions: Our experiments demonstrated a significant down regulation of connexin 43 by TGF-beta 1 in cultured hBSMCs. These findings support the view that TGF-beta 1 is involved in the pathophysiology of urinary bladder dysfunction. (C) 2008 European Association of Urology. Published by Elsevier B.V. All rights reserved.