Elevated polyamines in urothelial cells from OAB subjects mediate oxotremorine-evoked rapid intracellular calcium rise and delayed acetylcholine release.
Elevated polyamines in urothelial cells from OAB subjects mediate oxotremorine-evoked rapid intracellular calcium rise and delayed acetylcholine release.
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OAB 受试者尿路上皮细胞中多胺含量升高介导氧化震颤素引起的细胞内钙快速升高和乙酰胆碱释放延迟。
DOI：
10.1152/ajprenal.00345.2012
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
Chai,TobyC
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文献类型：
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作者：
Li,Mingkai;Sun,Yan;Tomiya,Noboru;Hsu,Yuchao;Chai,TobyC
膀胱尿路上皮细胞（布克）中多胺信号的增加可能在膀胱过度活动症（OAB）的病理生理学中起作用。我们定量了OAB和无症状（NB）受试者培养的布克细胞内多胺水平。我们评估了多胺是否调制快速细胞内钙（[Ca 2 +]i）的变化和延迟乙酰胆碱（ACh）释放诱发的oxotremorine（OXO，毒蕈碱激动剂）。从膀胱镜活检中培养布克。高效液相色谱法（HPLC）定量细胞内腐胺，亚精胺和精胺水平。五毫米二氟甲基鸟氨酸（DFMO），和一毫米甲基甘氨酸双胍酰腙（MGBG）的治疗被用来消耗细胞内的多胺。使用10微米的OXO来增加[Ca 2 +]水平（通过Fura 2微荧光测定法测量）并触发细胞外ACh释放（通过ELISA测量）。与NB布克C相比，OAB中的多胺水平升高（腐胺为0.5 ± 0.15 nmol/mg vs. 0.16 ± 0.03 nmol/mg，亚精胺为2.4 ± 0.21 nmol/mg vs. 1.01 ± 0.13 nmol/mg，精胺为1.90 ± 0.27 nmol/mg vs. 0.86 ± 0.26 nmol/mg;所有比较P< 0.05）。OXO使OAB组[Ca ~（2+）] i升高（205.10 ± 18.82%），NB布克组[Ca ~（2+）] i升高（119.54 ± 13.01%），差异有统计学意义（P< 0.05）。OAB和NB布克C中OXO诱发的[Ca ~（2+）] i升高分别为43.40 ± 6.45和38.82 ± 3.5%。OXO倾向于增加OAB与NB布克的ACh释放（分别为9.02 ± 0.1 vs. 7.04 ± 0.09 μM;P< 0.05）。多胺消耗减少ACh释放的OAB和NB布克。总之，OAB布克中多胺水平升高两倍。OXO引起OAB布克[Ca ~（2+）] i和ACh释放的增加，尽管这两个事件可能无关。多胺的耗尽导致OAB布克的行为类似于NB布克。
Increased polyamine signaling in bladder urothelial cells (BUC) may play a role in the pathophysiology of overactive bladder (OAB). We quantitated intracellular polyamine levels in cultured BUC from OAB and asymptomatic (NB) subjects. We assessed whether polyamines modulated rapid intracellular calcium ([Ca2+]i) changes and delayed acetylcholine (ACh) release evoked by oxotremorine (OXO, a muscarinic agonist). BUC were cultured from cystoscopic biopsies. High-performance liquid chromatography (HPLC) quantitated intracellular putrescine, spermidine, and spermine levels. Five-millimeter difluoromethylornithine (DFMO), and one-millimeter methylglyoxalbisguanylhydrazone (MGBG) treatments were used to deplete intracellular polyamines. Ten micrometers of OXO were used to increase [Ca2+]ilevels (measured by fura 2 microfluorimetry) and trigger extracellular ACh release (measured by ELISA). Polyamine levels were elevated in OAB compared with NB BUC (0.5 ± 0.15 vs. 0.16 ± 0.03 nmol/mg for putrescine, 2.4 ± 0.21 vs. 1.01 ± 0.13 nmol/mg for spermidine, and 1.90 ± 0.27 vs. 0.86 ± 0.26 nmol/mg for spermine;P< 0.05 for all comparisons). OXO evoked greater [Ca2+]irise in OAB (205.10 ± 18.82% increase over baseline) compared with in NB BUC (119.54 ± 13.01%;P< 0.05). After polyamine depletion, OXO evoked [Ca2+]irise decreased in OAB and NB BUC to 43.40 ± 6.45 and 38.82 ± 3.5%, respectively. OXO tended to increase ACh release by OAB vs. NB BUC (9.02 ± 0.1 vs. 7.04 ± 0.09 μM, respectively;P< 0.05). Polyamine depletion reduced ACh release by both OAB and NB BUC. In conclusion, polyamine levels were elevated twofold in OAB BUC. OXO evoked greater increase in [Ca2+]iand ACh release in OAB BUC, although these two events may be unrelated. Depletion of polyamines caused OAB BUC to behave similarly to NB BUC.