Abstract
Aging of the bladder detrusor muscle plays an important role in lower urinary tract symptoms in elderly people. Our previous work demonstrated that elderly patients have increased levels of vascular endothelial growth inhibitor (VEGI) in bladder tissue. Therefore, we hypothesized that VEGI may play a role in aging of the bladder detrusor muscle cells. This study aims to develop and characterize primary cultures of aged porcine bladder detrusor muscle cells in order to explore the expression and function of VEGI. Bladder samples from female pigs were divided into two groups: the aged group (Model) and the young group (Control). We confirmed β-galactosidase expression, a marker for senescence, in aged muscle cells (identified by α-smooth muscle actin (α-SMA) staining), but not in the young group. mRNA levels of VEGI-251 and death receptor 3 (DR3) were up-regulated (P < 0.05) and total cell protein levels of VEGI-251, DR3 and nuclear factor-kappa B [NF-κB (p65)], membrane protein levels of DR3, and nuclear protein levels of NF-κB (p65) were significantly higher (P < 0.01) in the Model cells compared to Control cells. In conclusion, we have established a method to culture aged detrusor muscle cells derived from porcine bladder. Higher levels of VEGI-251, DR3 and NF-κB (p65) were observed in the aged cells. VEGI-251 may function by increasing DR3 on cellular membranes and promoting the transfer of NF-κB into the nucleus. This suggests that VEGI may be a target for reversing the aging process of bladder detrusor muscle cells.
Similar content being viewed by others
References
Akino H, Chapple CR, McKay N, Cross RL, Murakami S, Yokoyama O, Chess-Williams R, Sellers DJ (2008) Spontaneous contractions of the pig urinary bladder: the effect of ATP-sensitive potassium channels and the role of the mucosa. BJU Int 102:1168–1174
Al-Lamki RS, Wang J, Tolkovsky AM, Bradley JA, Griffin JL, Thiru S, Wang EC, Bolton E, Min W, Moore P, Pober JS, Bradley JR (2008) TL1A both promotes and protects from renal inflammation and injury. J Am Soc Nephrol 19:953–960
Bamias G, Martin C III, Marini M, Hoang S, Mishina M, Ross WG, Sachedina MA, Friel CM, Mize J, Bickston SJ, Pizarro TT, Wei P, Cominelli F (2003) Expression, localization, and functional activity of TL1A, a novel Th1-polarizing cytokine in inflammatory bowel disease. J Immunol 171:4868–4874
Bamias G, Siakavellas SI, Stamatelopoulos KS, Chryssochoou E, Papamichael C, Sfikakis PP (2008) Circulating levels of TNF-like cytokine 1A (TL1A) and its decoy receptor 3 (DcR3) in rheumatoid arthritis. Clin Immunol 129:249–255
Cheng Y, Mansfield KJ, Sandow SL, Sadananda P, Burcher E, Moore KH (2011) Porcine bladder urothelial, myofibroblast, and detrusor muscle cells: characterization and ATP release. Front Pharmacol. doi:10.3389/fphar.2011.00027
Chew LJ, Pan H, Yu J, Tian S, Huang WQ, Zhang JY, Pang S, Li LY (2002) A novel secreted splice variant of vascular endothelial cell growth inhibitor. FASEB J 16:742–744
Cho H, Park SH, Lee S, Kang M, Hasty KA, Kim SJ (2011) Snapshot of degenerative aging of porcine intervertebral disc: a model to unravel the molecular mechanisms. Exp Mol Med 43:334–340
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O et al (1995) A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 92:9363–9367
Drake MJ, Fry CH, Eyden B (2006) Structural characterization of myofibroblasts in the bladder. BJU Int 97:29–32
Duan L, Yang G, Zhang R, Feng L, Xu C (2012) Advancement in the research on vascular endothelial growth inhibitor (VEGI). Targeted Oncol 7:87–90
Ge Z, Sanders AJ, Ye L, Jiang WG (2011) Aberrant expression and function of death receptor-3 and death decoy receptor-3 in human cancer. Exp Ther Med 2:167–172
Gillespie JI, Markerink-van Ittersum M, De Vente J (2006) Endogenous nitric oxide/cGMP signalling in the guinea pig bladder: evidence for distinct populations of sub-urothelial interstitial cells. Cell Tissue Res 325:325–332
Hashitani H, Brading AF (2003) Electrical properties of detrusor smooth muscles from the pig and human urinary bladder. Br J Pharmacol 140:146–158
Hernández M, Knight GE, Wildman SS, Burnstock G (2009) Role of ATP and related purines in inhibitory neurotransmission to the pig urinary bladder neck. Br J Pharmacol 157:1463–1473
Hou W, Medynski D, Wu S, Lin X, Li LY (2005) VEGI-192, a new isoform of TNFSF15, specifically eliminates tumor vascular endothelial cells and suppresses tumor growth. Clin Cancer Res 11:5595–5602
Itahana K, Campisi J, Dimri GP (2007) Methods to detect biomarkers of cellular senescence: the senescence-associated beta-galactosidase assay. Methods Mol Biol 371:21–31
Iwanaga K, Murata T, Hori M, Ozaki H (2010) Isolation and characterization of bovine intestinal subepithelial myofibroblasts. J Pharmacol Sci 112:98–104
Jin T, Kim S, Guo F, Howard A, Zhang YZ (2007) Purification and crystallization of recombinant human TNF-like ligand TL1A. Cytokine 40:115–122
Johnston L, Woolsey S, Cunningham RM, O’Kane H, Duggan B, Keane P, McCloskey KD (2010) Morphological expression of KIT positive interstitial cells of Cajal in human bladder. J Urol 184:370–377
Kitson J, Raven T, Jiang YP, Goeddel DV, Giles KM, Pun KT, Grinham CJ, Brown R, Farrow SN (1996) A death-domain-containing receptor that mediates apoptosis. Nature 384:372–375
Kumar V, Chapple CC, Chess-Williams R (2004) Characteristics of adenosine triphosphate [corrected] release from porcine and human normal bladder. J Urol 172:744–747
Lan W, Petznick A, Heryati S, Rifada M, Tong L (2012) Nuclear Factor-kappaB: central regulator in ocular surface inflammation and diseases. Ocul Surf 10:137–148
Lee YS, Lee KS, Jung JH, Han DH, Oh SJ, Seo JT, Lee JG, Park HS, Choo MS (2011) Prevalence of overactive bladder, urinary incontinence, and lower urinary tract symptoms: results of Korean EPIC study. World J Urol 29:185–190
Marsters SA, Sheridan JP, Donahue CJ, Pitti RM, Gray CL, Goddard AD, Bauer KD, Ashkenazi A (1996) Apo-3, a new member of the tumor necrosis factor receptor family, contains a death domain and activates apoptosis and NF-kappa B. Curr Biol 6:1669–1676
McCloskey KD (2010) Interstitial cells in the urinary bladder–localization and function. Neurol Urodyn 29:82–87
Migone TS, Zhang J, Luo X, Zhuang L, Chen C, Hu B, Hong JS, Perry JW, Chen SF, Zhou JX, Cho YH, Ullrich S, Kanakaraj P, Carrell J, Boyd E, Olsen HS, Hu G, Pukac L, Liu D, Ni J, Kim S, Gentz R, Feng P, Moore PA, Ruben SM, Wei P (2002) TL1A is a TNF-like ligand for DR3 and TR6/DcR3 and functions as a T cell costimulator. Immunity 16:479–492
Prehn JL, Mehdizadeh S, Landers CJ, Luo X, Cha SC, Wei P, Targan SR (2004) Potential role for TL1A, the new TNF-family member and potent costimulator of IFN-gamma, in mucosal inflammation. Clin Immunol 112:66–77
Sadananda P, Chess-Williams R, Burcher E (2008) Contractile properties of the pig bladder mucosa in response to neurokinin A: a role for myofibroblasts? Br J Pharmacol 153:1465–1473
Severino J, Allen RG, Balin S, Balin A, Cristofalo VJ (2000) Is beta-galactosidase staining a marker of senescence in vitro and in vivo? Exp Cell Res 257:162–171
Sikora E, Arendt T, Bennett M, Narita M (2011) Impact of cellular senescence signature on ageing research. Ageing Res Rev 10:146–152
Southgate J, Harnden P, Trejdosiewicz LK (1999) Cytokeratin expression patterns in normal and malignant urothelium: a review of the biological and diagnostic implications. Histol Histopathol 14:657–664
Srivastava SK, Ramana KV (2009) Focus on molecules: nuclear factor-kappaB. Exp Eye Res 88:2–3
Sui GP, Wu C, Fry CH (2006) Characterization of the purinergic receptor subtype on guinea-pig suburothelial myofibroblasts. BJU Int 97:1327–1331
Tan KB, Harrop J, Reddy M, Young P, Terrett J, Emery J, Moore G, Truneh A (1997) Characterization of a novel TNF-like ligand and recently described TNF ligand and TNF receptor superfamily genes and their constitutive and inducible expression in hematopoietic and non-hematopoietic cells. Gene 204:35–46
Templeman L, Sellers DJ, Chapple CR, Rosario DJ, Hay DP, Chess-Williams R (2003) Investigation of neurokinin-2 and -3 receptors in the human and pig bladder. BJU Int 92:787–792
Trifillis AL, Cui X, Jacobs S, Warren JW (1995) Culture of bladder epithelium from cystoscopic biopsies of patients with interstitial cystitis. J Urol 153:243–248
Vandesompele J, De Preter K, Pattyn F, PoPPe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:34.1–34.11
Wang W, Zhang N, Zhu XH, He ZS, Wahafu W, Xu ZQ, Yang Y (2012) Involvement of TL1A and DR3 in induction of ischaemia and inflammation in urinary bladder dysfunction in the elderly. Mol Med Rep 6:434–438
Welch LC, Taubenberger S, Tennstedt SL (2011) Patients’ experiences of seeking health care for lower urinary tract symptoms. Res Nursing Health 34:496–507
Wong ET, Tergaonkar V (2009) Roles of NF-kappaB in health and disease: mechanisms and therapeutic potential. Clin Sci 116:451–465
Wood DN, Brown RA, Fry CH (2004) Characterization of the control of intracellular [Ca2+] and the contractile phenotype of cultured human detrusor smooth muscle cells. J Urol 172:753–757
Wu C, Sui GP, Fry CH (2004) Purinergic regulation of guinea pig suburothelial myofibroblasts. J Physiol 559:231–243
Yao JJ, Zhang M, Miao XH, Zhao P, Zhu SY, Ding H, Qi ZT (2006) Isoform of vascular endothelial cell growth inhibitor (VEGI72-251) increases interleukin-2 production by activation of T lymphocytes. Acta Biochim Biophys Sin 38:249–253
Zhai Y, Ni J, Jiang GW, Lu J, Xing L, Lincoln C, Carter KC, Janat F, Kozak D, Xu S, Rojas L, Aggarwal BB, Ruben S, Li LY, Gentz R, Yu GL (1999) VEGI, a novel cytokine of the tumor necrosis factor family, is an angiogenesis inhibitor that suppresses the growth of colon carcinomas in vivo. FASEB J 13:181–189
Zhang N, Sanders AJ, Ye L, Jiang WG (2009) Vascular endothelial growth inhibitor in human cancer (Review). Int J Mol Med 24:3–8
Zhang E, Zhu X, Han S, Peng Z, Wang W, Li J, Yang Y (2013) Increased expression of TNF ligand-related molecule 1A and death receptor 3 in bladder tissues of patients with painful bladder syndrome/interstitial cystitis. Exp Ther Med 5:282–286
Acknowledgments
This study was supported by grants from National Natural Science Foundation of China (No. 81070603). The authors particularly appreciate the valuable comments from all other members in their laboratory.
Conflict of interest
All authors have no conflicts of interest to disclose.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, M., Zhang, N., Wang, W. et al. Expression and function of vascular endothelial growth inhibitor in aged porcine bladder detrusor muscle cells. Biogerontology 14, 543–556 (2013). https://doi.org/10.1007/s10522-013-9460-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10522-013-9460-5