Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA.
Chem Biol Drug Des. 2011 Jun;77(6):421-30. doi: 10.1111/j.1747-0285.2011.01108.x. Epub 2011 Apr 27.
Interstitial cystitis/painful bladder syndrome is a chronic bladder disorder with epithelial thinning or ulceration, pain, urinary frequency and urgency, for which there is no reliably effective therapy. We previously reported that interstitial cystitis/painful bladder syndrome bladder epithelial cells make a glycopeptide antiproliferative factor or 'APF' (Neu5Acα2-3Galβ1-3GalNAcα-O-TVPAAVVVA) that induces abnormalities in normal cells similar to those in interstitial cystitis/painful bladder syndrome cells in vitro, including decreased proliferation, decreased tight junction formation, and increased paracellular permeability. We screened inactive APF derivatives for their ability to block antiproliferative activity of asialylated-APF ('as-APF') in normal bladder cells and determined the ability of as-APF-blocking derivatives to normalize tight junction protein expression, paracellular permeability, and/or proliferation of interstitial cystitis/painful bladder syndrome cells. Only two of these derivatives [Galβ1-3GalNAcα-O-TV-(d-pipecolic acid)-AAVVVA and Galβ1-3GalNAcα-O-TV-(d-proline)-AAVVVA] blocked as-APF antiproliferative activity in normal cells (p < 0.001 for both). Both of these antagonists also 1) significantly increased mRNA expression of ZO-1, occludin, and claudins 1, 4, 8, and 12 in interstitial cystitis/painful bladder syndrome cells by qRT-PCR; 2) normalized interstitial cystitis/painful bladder syndrome epithelial cell tight junction protein expression and tight junction formation by confocal immunofluorescence microscopy; and 3) decreased paracellular permeability of (14) C-mannitol and (3) H-inulin between confluent interstitial cystitis/painful bladder syndrome epithelial cells on Transwell plates, suggesting that these potent APF antagonists may be useful for the development as interstitial cystitis/painful bladder syndrome therapies.
间质性膀胱炎/膀胱疼痛综合征是一种慢性膀胱疾病，表现为上皮变薄或溃疡、疼痛、尿频和尿急，目前尚无可靠有效的治疗方法。我们之前报道过，间质性膀胱炎/膀胱疼痛综合征膀胱上皮细胞产生糖肽增殖抑制因子或“APF”（Neu5Acα2-3Galβ1-3GalNAcα-O-TVPAAVVVA），在体外诱导正常细胞出现类似于间质性膀胱炎/膀胱疼痛综合征细胞的异常，包括增殖减少、紧密连接形成减少和细胞旁通透性增加。我们筛选了无活性的 APF 衍生物，以评估它们阻断正常膀胱细胞中去唾液酸化-APF（“as-APF”）增殖抑制活性的能力，并确定了 as-APF 阻断衍生物使间质性膀胱炎/膀胱疼痛综合征细胞的紧密连接蛋白表达、细胞旁通透性和/或增殖正常化的能力。只有两种此类衍生物[Galβ1-3GalNAcα-O-TV-(d-哌啶酸)-AAVVVA 和 Galβ1-3GalNAcα-O-TV-(d-脯氨酸)-AAVVVA]能阻断正常细胞中 as-APF 的增殖抑制活性（两者均 p < 0.001）。这两种拮抗剂还 1）通过 qRT-PCR 显著增加了间质性膀胱炎/膀胱疼痛综合征细胞中 ZO-1、occludin 和 Claudin 1、4、8 和 12 的 mRNA 表达；2）通过共聚焦免疫荧光显微镜使间质性膀胱炎/膀胱疼痛综合征上皮细胞的紧密连接蛋白表达和紧密连接形成正常化；3）减少了 Transwell 板上间质性膀胱炎/膀胱疼痛综合征上皮细胞之间的（14）C-甘露醇和（3）H-尿囊素的细胞旁通透性，表明这些有效的 APF 拮抗剂可能可用于开发间质性膀胱炎/膀胱疼痛综合征的治疗方法。
