内皮素-1在兔膀胱出口部分梗阻继发膀胱增生中的可能作用。

Possible role of endothelin-1 in the rabbit urinary bladder hyperplasia secondary to partial bladder outlet obstruction.

作者信息

Khan M A, Shukla N, Auld J, Thompson C S, Mumtaz F H, Stansby G P, Morgan R J, Mikhailidis D P

机构信息

Department of Urology, Royal Free and University College Medical School, University College London, UK.

出版信息

Scand J Urol Nephrol. 2000 Feb;34(1):15-20. doi: 10.1080/003655900750016823.

DOI:10.1080/003655900750016823

PMID:10757264

Abstract

OBJECTIVES

Urinary bladder hypertrophy and hyperplasia are common features of bladder outlet obstruction (BOO). The urinary bladder is known to synthesize endothelin-1 (ET-1), which is a potent vasoconstrictor peptide with mitogenic properties. Using an animal model of partial BOO, we investigated the potential role of ET-1 and its receptor subtypes (ET(A) and ET(B)) in bladder smooth muscle cell (SMC) proliferation.

MATERIALS AND METHODS

Partial BOO was produced in adult male New Zealand White rabbits. After 3 weeks, the bladder was removed and SMCs from the dome and bladder neck were grown using standard explant methodology. At passage 2, the cells were made quiescent and then further incubated in foetal calf serum (FCS), control age-matched rabbit serum (CRS) or partial BOO serum (BRS) in the presence or absence of ET(A)-antagonist (BQ123) or ET(B)-antagonist (BQ788). SMC proliferation was then measured 24 h later with 5-bromo-2'deoxy-uracil and by cell counting using a haemocytometer at 48 h. Immunostaining for alpha-actin was performed on detrusor and bladder neck cells to confirm the presence of smooth muscle cells.

RESULTS

BQ123 and BQ788 did not influence detrusor or bladder neck SMC proliferation in FCS or CRS. However, in the presence of BRS, BQ123 and BQ788 (100 nmol/L) significantly (p = 0.008) inhibited detrusor and bladder neck SMC proliferation. Cell counts were significantly reduced from the detrusor (p = 0.03, p = 0.01 with BQ123 and BQ788, respectively) and bladder neck (p = 0.01 for both BQ123 and BQ78).

CONCLUSIONS

These results suggest that ET antagonists may have a role in preventing SMC hyperplasia associated with partial BOO.

摘要

目的

膀胱肥大和增生是膀胱出口梗阻（BOO）的常见特征。已知膀胱能合成内皮素-1（ET-1），它是一种具有促有丝分裂特性的强效血管收缩肽。我们使用部分BOO动物模型，研究ET-1及其受体亚型（ET(A)和ET(B)）在膀胱平滑肌细胞（SMC）增殖中的潜在作用。

材料与方法

在成年雄性新西兰白兔中制造部分BOO。3周后，取出膀胱，使用标准外植体方法培养来自膀胱顶部和膀胱颈部的SMC。在第2代时，使细胞静止，然后在存在或不存在ET(A)拮抗剂（BQ123）或ET(B)拮抗剂（BQ788）的情况下，在胎牛血清（FCS）、对照年龄匹配的兔血清（CRS）或部分BOO血清（BRS）中进一步孵育。然后在24小时后用5-溴-2'-脱氧尿嘧啶测量SMC增殖，并在48小时时使用血细胞计数器进行细胞计数。对逼尿肌和膀胱颈部细胞进行α-肌动蛋白免疫染色，以确认平滑肌细胞的存在。

结果

BQ123和BQ788在FCS或CRS中不影响逼尿肌或膀胱颈部SMC增殖。然而，在存在BRS的情况下，BQ123和BQ788（100 nmol/L）显著（p = 0.008）抑制逼尿肌和膀胱颈部SMC增殖。逼尿肌的细胞计数显著减少（BQ123和BQ788分别为p = 0.03，p = 0.01），膀胱颈部的细胞计数也显著减少（BQ123和BQ78均为p = 0.01）。