Bing Wu, Chang Shaohua, Hypolite Joseph A, DiSanto Michael E, Zderic Stephen A, Rolf Lester, Wein Alan J, Chacko Samuel
Department of Pathobiology, University of Pennsylvania, Philadelphia 19104, USA.
Am J Physiol Renal Physiol. 2003 Nov;285(5):F990-7. doi: 10.1152/ajprenal.00378.2002. Epub 2003 Jul 8.
Detrusor smooth muscle (DSM) undergoes hypertrophy after partial bladder outlet obstruction (PBOO) in male rabbits, as it does in men with PBOO induced by benign prostatic hyperplasia. Despite detrusor hypertrophy, some bladders are severely dysfunctional (decompensated). In this study, the rabbit model for PBOO was used to determine the biochemical regulation of the contractile apparatus and force maintenance by the detrusor from decompensated bladders (DB). Bladders from sham-operated rabbits served as a control. On stimulation with 125 mM KCl, the DSM from sham-operated (SB) rabbits showed phasic contractions, whereas the detrusor from DB was tonic, exhibiting slow development of force, a longer duration of force maintenance, and slow relaxation. The Rho kinase (ROK) inhibitor Y-27632 enhanced the relaxation of precontracted DSM strips from DB. The enhancement of relaxation of the KCl-induced contraction of DB by Y-27632 was associated with dephosphorylation of myosin light chain (MLC20). The DSM extract from DB showed low phosphatase activity compared with that from SB. The DB also showed more Ca2+-independent MLC20 phosphorylation, which was partially inhibited by Y-27632. RT-PCR and Western blotting revealed similar expression levels of MLC kinase and ROK-alpha in SB and DB, but ROK-beta was overexpressed in DB. These results suggest that the ROK-mediated pathway is partly responsible for the high degree of force maintenance and slow relaxation in the detrusor from DB.
雄性兔部分膀胱出口梗阻(PBOO)后,逼尿肌平滑肌(DSM)会发生肥大,这与良性前列腺增生所致PBOO的男性情况相同。尽管存在逼尿肌肥大,但一些膀胱功能严重失调(失代偿)。在本研究中,采用PBOO兔模型来确定失代偿膀胱(DB)逼尿肌收缩装置的生化调节及力量维持情况。假手术兔的膀胱作为对照。用125 mM KCl刺激时,假手术(SB)兔的DSM呈现相性收缩,而DB的逼尿肌呈强直性收缩,表现为力量缓慢发展、力量维持时间更长以及松弛缓慢。Rho激酶(ROK)抑制剂Y-27632增强了DB预收缩DSM条带的松弛。Y-27632增强DB的KCl诱导收缩的松弛与肌球蛋白轻链(MLC20)的去磷酸化有关。与SB相比,DB的DSM提取物显示出较低的磷酸酶活性。DB还显示出更多的不依赖Ca2+的MLC20磷酸化,Y-27632可部分抑制这种磷酸化。逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法显示,SB和DB中MLC激酶和ROK-α的表达水平相似,但DB中ROK-β过表达。这些结果表明,ROK介导的途径部分导致了DB逼尿肌高度的力量维持和缓慢松弛。