The role of autonomic nerve receptor subtypes and G-proteins for the cause of overactive bladder

毒蕈碱受体亚型在介导逼尿肌收缩中的作用：使用尿道的纵向和横向条，在正常和cAMP升高的组织中计算毒蕈碱拮抗剂对卡巴胆碱反应的亲和力（用KCl收缩，用异丙肾上腺素舒张）。猪尿道似乎主要具有M_2毒蕈碱受体。正常尿道的收缩主要由环肌的M_2和M_3受体介导，而由纵肌的M_3受体介导。cAMP升高后，M_2受体参与膀胱和尿道平滑肌的收缩，但环肌中M_2受体的参与更大。从猪的逼尿肌和尿道肌制备细胞膜，并与[^3H]-二氢烯丙诺 ...更多信息 用非选择性β-肾上腺素受体拮抗剂DHA 1作为特异性放射配体，测定β-肾上腺素受体亚型的存在。在功能实验中，分离的逼尿肌肌条用于确定激动剂的效力和拮抗剂的亲和力。β_3-肾上腺素受体是猪膀胱和尿道中的主要β-肾上腺素受体。在功能上，该组织的β-肾上腺素受体介导的反应是通过β_2和β_3肾上腺素受体亚型介导的。β_3-肾上腺素受体在猪和人的膀胱体中似乎更多地参与舒张，而β_2-肾上腺素受体在猪的三角区和尿道平滑肌中似乎更多地参与舒张。Rho/Rho激酶主要存在于控制离体人膀胱收缩的尿道平滑肌中：从接受膀胱癌全膀胱切除术的患者获得来自人膀胱穹隆区域的逼尿肌样品。采用多重RT-PCR和Western blotting方法检测Rho激酶的mRNA和蛋白水平。使用与用于Western印迹的抗体相同的特异性抗体进行免疫组织化学，以支持通过Western印迹获得的数据。获得卡巴胆碱的累积浓度-响应曲线（CRC），然后用含10 μ M Y 27632或溶剂（时间对照）的Krebs溶液平衡30 min。在相对多重RT-PCR中，RhoA和Rho激酶在尿道平滑肌中的含量高于在尿道平滑肌中的含量。蛋白质印迹法也证实Rho激酶蛋白在尿路上皮中的表达显著高于平滑肌（分别为p <0.0001和p <0.0001）。免疫组化染色显示，尿路上皮细胞对Rho激酶抗体呈中至强反应，平滑肌细胞呈弱至中度反应。Rho和Rho激酶似乎主要存在于尿路上皮中。最大反应卡巴胆碱的存在下，尿路刺激减少。Rho/Rho激酶途径似乎参与了人膀胱平滑肌在尿激酶存在下的收缩。少

The role of muscarinic receptor subtypes in mediating contraction of detrusor muscles : The affinities of muscarinic antagonists against carbachol responses were calculated in normal and cAMP-elevated tissues (contraction with KCl and relaxation with isoprenaline) using longitudinal and transverse strips of urethra. The pig urethra appears to possess predominantly M_2 muscarinic receptors. Contraction of the normal urethra appears to be predominantly mediated by M_2 and M_3-receptors in circular muscle, but by M_3-receptors in longitudinal muscle. Following cAMP-elevation, a contribution to contraction of M_2-receptors appeared to be demonstrated in both tissues, but the involvement of the M_2-receptors appeared greater in the circular muscle.The role of β_3-Adrenoceptors in mediating relaxation of bladder and urethral smooth muscle : of the pig and human. : Cell membranes were prepared from detrusor and urethral muscle of the pig and competition experiments with [^3H]-dihydroalprenolo … More l(DHA), a non-selective β-adrenoceptor antagonist was used as a specific radioligand to determine the presence of β-adrenoceptor subtypes. In functional experiments, isolated detrusor muscle strips were used to determine the potency of agonists and the affinity of antagonists. β_3-adrenoceptars are the predominant β-adrenoceptor subtype present in the pig bladder and urethra. Functionally, β-adrenoceptor mediated responses of this tissue are mediated via both the β_2- and β_3-adrenoceptor subtypes. β_3-adrenoceptors appear to be more involved in relaxation of bladder body in the pig and human, whereas β_2-adrenoceptors appear to be more involved in trigonal and urethral smooth muscle of the pig.Rho/Rho kinase is predominantly present in the urothelium controlling the contraction of the human urinary bladder in vitro : Samples of detrusor muscle from the dome region of the human urinary bladder were obtained from patients undergoing total cystectomy for bladder cancer. The mRNA and protein levels of Rho-kinase were analyzed using the relative multiplex RT-PCR and Western blotting. Immunohistochemistry, using the same specific antibodies as antibodies for Western blotting, was performed to support the data obtained by Western blotting. Cumulative concentration-response curves(CRCs) to carbachol were obtained, and were then equilibrated for 30 mm with Krebs solution containing the 10μM of Y 27632 or vehicle (time control). On the relative multiplex RT-PCR,RhoA and Rho kinase were more abundant in the urothelium than in the smooth muscle. Western blotting also demonstrated significantly greater expression of Rho kinase protein expression in the urothelium than in the smooth muscle (p<0.0001 and p<0.0001, respectively). Urothelial cells showed moderate to strong reaction for Rho kinase antibodies, and smooth muscle cells showed weak to moderate reaction on immunohistochemistrical study. Rho and Rho kinase appear to be predominantly present in the urothelium. The maximum response to carbachol was decreased in the presence of urothelium. The Rho/Rho kinase pathway appears to be involved in the contraction of human bladder smooth muscle in the presence of urothelium. Less