Expression of sodium/calcium exchanger subtype 2 and its physiological role in detrusor cells of rats

Xiao ZHONG; Pingxian WANG; Wengang HU; Qingqing WANG; Longkun LI; Chibing HUANG

doi:10.16016/j.1000-5404.201710150

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Expression of sodium/calcium exchanger subtype 2 and its physiological role in detrusor cells of rats

更新时间：2026-03-02

- Expression of sodium/calcium exchanger subtype 2 and its physiological role in detrusor cells of rats
- Journal of Third Military Medical University Vol. 40, Issue 4, Pages: 302-306(2018)
- 作者机构：
  
  1..400037 重庆，陆军军医大学(第三军医大学)第二附属医院：泌尿外二科First Department of Urology, Army Medical University (Third Military Medical University), Chongqing, 400037, China
  2..400037 重庆，陆军军医大学(第三军医大学)第二附属医院：泌尿外一科Second Department of Urology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation for Young Scholars of China(81400761)
- DOI：10.16016/j.1000-5404.201710150
  CLC： R322.62; R329-33; R329.27
- Received：23 October 2017，
  
  Published：28 February 2018
- 稿件说明：
移动端阅览
Xiao ZHONG, Pingxian WANG, Wengang HU, et al. Expression of sodium/calcium exchanger subtype 2 and its physiological role in detrusor cells of rats[J]. Journal of Third Military Medical University, 2018, 40(4): 302-306.
DOI：

Xiao ZHONG, Pingxian WANG, Wengang HU, et al. Expression of sodium/calcium exchanger subtype 2 and its physiological role in detrusor cells of rats[J]. Journal of Third Military Medical University, 2018, 40(4): 302-306. DOI： 10.16016/j.1000-5404.201710150.

摘要

目的

检测大鼠逼尿肌细胞是否表达钠钙交换体亚型2(NCX2)，并测试其功能。

方法

10只大鼠(7只雌性，3只雄性)为实验对象，取膀胱组织为实验组，大脑组织为对照组。首先利用急性酶分离技术，制备和培养大鼠逼尿肌细胞，再采用反转录-聚合酶链式反应和免疫印迹技术，检测大鼠膀胱组织中是否表达NCX2基因及蛋白；免疫荧光技术进一步明确NCX2与逼尿肌细胞的关系；膜片钳技术测试逼尿肌细胞是否存在NCX2电流。

结果

首先制备了光镜下以梭形为主，免疫组化染色α-actin呈阳性的逼尿肌细胞；利用逆转录-聚合酶链式反应，在大鼠大脑组织、膀胱组织中分别发现NCX2 mRNA表达；免疫印迹技术在上述组织中检测NCX2的蛋白相对表达量，差异均未见统计学意义；免疫荧光双抗进一步证实NCX2蛋白与逼尿肌细胞共表达；膜片钳检测到逼尿肌细胞上的NCX2电流，电流密度为(0.79±0.45) pA/pF。

结论

发现NCX2表达于大鼠逼尿肌细胞，电生理证实NCX2具有维持逼尿肌细胞内钙离子平衡的能力，其生理作用需要进一步的探索。

Abstract

Objective

To investigate the expression of sodium/calcium exchanger subtype 2 (NCX2) and explore its physiological role in detrusor cells of rats.

Methods

The bladder tissue was taken from 7 female and 3 male Wistar rats with the brain tissue as the control

and the detrusor cells were isolated from the bladder tissue using acute enzyme digestion technique. RT-PCR and Western blotting were used to detect the expression of NCX2 at mRNA and protein respectively in the bladder and brain tissues of the rats. Double-label immunofluorescence assay was used to verify the localization of NCX2 expression in the detrusor cells. NCX2 current was recorded using the whole-cell patch-clamp technique on the detrusor cells in the presence of the antagonist KB-R7943 (5 μmol/L).

Results

The acutely isolated detrusor cells showed predominantly a spindle-shaped morphology under microscope and were immunocytochemically positive for α-actin. NCX2 expression was detected in both the bladder and brain tissues of the rats with similar expression levels between the 2 tissues. Double-label immunofluorescence confirmed that NCX2 expression was localized on the detrusor cells

and NCX2 currents were detected in the detrusor cells with a current density of 0.79±0.45 pA/pF.

Conclusion

NCX2 is expressed on the detrusor cells of rats and the electrophysiological evidence obtained suggests that NCX2 contributes to the maintenance of intracellular calcium homeostasis in the detrusor cells

but its exact physiological role still needs further investigation.

关键词

Keywords

references

IRWIN D E, MILSOM I, HUNSKAAR S, et al . Population-based survey of urinary incontinence, overactive bladder, and other lower urinary tract symptoms in five countries: results of the EPIC study[J ] . Eur Urol, 2006, 50(6): 1306-1314. DOI: 10.1016/j.eururo.2006.09.019.

SUI G, FRY C H, MALONE-LEE J, et al . Aberrant Ca 2+ oscillations in smooth muscle cells from overactive human bladders[J ] . Cell Calcium, 2009, 45(5): 456-464. DOI: 10.1016/j.ceca.2009.03.001.

FRY C H, JABR R I. T-type Ca 2+ channels and the urinary and male genital tracts[J ] . Pflugers Arch, 2014, 466(4): 781-789. DOI: 10.1007/s00424-014-1446-x.

ZHAO B, ZHONG X, BAI X, et al . Changes in store-operated calcium channels in rat bladders with detrusor overactivity[J ] . Urology, 2014, 84(2): 491-491. DOI: 10.1016/j.urology.2014.05.007.

ZHONG X, DENG J, HE P, et al .Reverse mode of the sodium/calcium exchanger subtype 3 in interstitial cells of Cajal from rat bladder[J ] . Urology, 2013, 82(1): 254-257. DOI: 10.1016/j.urology.2013.02.049.

RACHANENI S, ARYA P, LATTHE P. Urinary nerve growth factor: a biomarker of detrusor overactivity? A systematic review[J]. Int Urogynecol J, 2013, 24(10): 1603-1609. DOI: 10.1007/s00192-013-2104-0.

JUSZCZAK K, MACIUKIEWICZ P, DREWA T, et al . Cajal-like interstitial cells as a novel target in detrusor overactivity treatment: true or myth?[J ] . Cent European J Urol, 2014, 66(4): 413-417. DOI: 10.5173/ceju.2013.04.art5.

MANSFIELD K J, LIU L, MOORE K H, et al . Molecular characterization of M2 and M3 muscarinic receptor expression in bladder from women with refractory idiopathic detrusor overactivity[J ] . BJU Int, 2007, 99(6): 1433-1438. DOI: 10.1111/j.1464-410X.2007.06866.x.

DENG J, HE P, ZHONG X, et al . Identification of T-type calcium channels in the interstitial cells of Cajal in rat bladder[J ] . Urology, 2012, 80(6): 1389.e1-7. DOI: 10.1016/j.urology.2012.07.031.

SAITO M, MIYAGAWA I. Bladder dysfunction after acute urinary retention in rats[J]. J Urol, 2001, 165(5): 1745-1747. DOI: 10.1097/00005392-200105000-00093.

李文刚, 方强, 杨景, 等.活性氧在急性尿潴留大鼠膀胱功能损害中的作用研究[J].第三军医大学学报, 2005, 27(4): 331-333. DOI: 10.3321/j.issn:1000-5404.2005.04.015.

LI W G, FANG Q, YANG J, et al . Roles of reactive oxygen species in acute retention bladder in rats[J ] . J Third Mil Med Univ, 2005, 27(4): 331-333. DOI:10.3321/j.issn:1000-5404.2005.04.015.

李文刚, 方强, 卢根生, 等.急性尿潴留大鼠膀胱组织黄嘌呤氧化还原酶活性变化及其意义[J].第三军医大学学报, 2006, 28(9): 973-975. DOI: 10.3321/j.issn:1000-5404.2006.09.033.

LI W G, FANG Q, LU G S, et al . Changes of xod activity induced by acute urinary retention in rats[J ] . J Third Mil Med Univ, 2006, 28(9): 973-975. DOI: 10.3321/j.issn:1000-5404.2006.09.033.

LIN A T, CHEN K K, YANG C H, et al . Mannitol facilitates rabbit urinary bladder recovery from overdistension injury[J ] . Urology, 2000, 56(4): 702-707.

TUO Q R, MA Y F, CHEN W, et al . Reactive oxygen species induce a Ca 2+ -spark increase in sensitized murine airway smooth muscle cells[J ] . Biochem Biophys Res Commun, 2013, 434(3):498-502. DOI: 10.1016/j.bbrc.2013.03.102.

SHATTOCK M J, OTTOLIA M, BERS D M, et al . Na + /Ca 2+ exchange and Na + /K + -ATPase in the heart[J ] . J Physiol, 2015, 593(6): 1361-1382. DOI: 10.1113/jphysiol.2014.282319.

ZHANG S, DONG H, RUBIN LJ, et al . Upregulation of Na + /Ca 2+ exchanger contributes to the enhanced Ca 2+ entry in pulmonary artery smooth muscle cells from patients with idiopathic pulmonary arterial hypertension[J ] . Am J Physiol, Cell Physiol, 2007, 292(6): C2297-C2305. DOI: 10.1152/ajpcell.00383.2006.

NISHIYAMA K, AZUMA Y T, MORIOKA A, et al . Roles of Na + /Ca 2+ exchanger isoforms NCX1 and NCX2 in motility in mouse ileum[J ] . Naunyn Schmiedebergs Arch Pharmacol, 2016, 389(10):1081-1090. DOI: 10.1007/s00210-016-1271-1.

KHAKSAR S, BIGDELI M R. Anti-excitotoxic effects of cannabidiol are partly mediated by enhancement of NCX2 expression in animal model of cerebral ischemia[J]. Eur J Pharmacol, 2017, 794: 270-279. DOI: 10.1016/j.ejphar.2016.11.011.

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