Study on the Relationship between Pressure and Cell Injury of Human Bladder Smooth Muscle Cells_West China Medical Journal

Authors：

 WEIXin , ZHOULiang , WANGKun-jie , LIHong

Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding author：

Keywords：

Pressure; Bladder smooth muscle cells; Injury

DOI：

10.7507/1002-0179.20150310

Video：

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Objective To explore the relationship between the pressure level within the scope of promoting proliferation and cell injury of human bladder smooth muscle cells (HBSMCs). Method HBSMCs in vitro were divided into the experimental group and control group. The cells in the experimental group were exposed to 40 cm H₂O (1 cm H₂O=0.098 kPa) pressure and those in the control group were cultured in normal condition for 24 hours. We investigated the cell morphology and cytoskeleton with indirect immunofluorescence staining for α-actin. Propidium iodide (PI) staining was applied to evaluate the level of cell apoptosis. Results There was no significant difference in the cell morphology between the two groups. However, the expression of α-actin in the experimental group[(50.93±1.99)%] was significantly reduced comparing with that in the control group[(24.70±1.61)%] (t=32.404, P<0.001). The results of PI staining showed that compared with the control group[(3.50±2.12)%], the number of PI staining positive cells in the experimental group [(9.00±1.41)%] was significantly higher (t=6.110, P<0.001). Conclusions Pressure condition can promotes cell proliferation, but at the same time, it can also lead to cell injury of HBSMCs.

Citation： WEIXin, ZHOULiang, WANGKun-jie, LIHong. Study on the Relationship between Pressure and Cell Injury of Human Bladder Smooth Muscle Cells. West China Medical Journal, 2015, 30(6): 1079-1082. doi: 10.7507/1002-0179.20150310 Copy

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8.	Vouyouka AG, Jiang Y, Rastogi R, et al. Ambient pressure upregulates nitric oxide synthase in a phosphorylated-extracellular regulated kinase-and protein kinase C-dependent manner[J]. J Vasc Surg, 2006, 44(5):1076-1084.
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13.	Butler DL, Goldstein S, Guilak F. Functional tissue engineering:the role of biomechanics[J]. J Biomech Eng, 2000, 122(6):570-575.
14.	Kaspar D, Seidl W, Neidlinger-Wilke C, et al. Proliferation of human-derived osteoblast-like cells depends on the cycle number and frequency of uniaxial strain[J]. J Biomech, 2002, 35(7):873-880.
15.	Leung DY, Glagov S, Mathews MB. Cyclic stretching stimulates synthesis of matrix components by arterial smooth muscle cells in vitro[J]. Science, 1976, 191(4226):475-477.
16.	Houtchens GR, Foster MD, Desai TA, et al. Combined effects of microtopography and cyclic strain on vascular smooth muscle cell orientation[J]. J Biomech, 2008, 41(4):762-769.
17.	Shah JB, Mcconkey DJ, Dinney CP. New strategies in muscle-invasive bladder cancer:on the road to personalized medicine[J]. Clin Cancer Res, 2011, 17(9):2608-2612.
18.	Brown AL, Brook-Allred TT, Waddell JE, et al. Bladder acellular matrix as a substrate for studying in vitro bladder smooth muscle-urothelial cell interactions[J]. Biomaterials, 2005, 26(5):529-543.
19.	Lee SD, Misseri R, Akbal C, et al. Muscarinic receptor expression increases following exposure to intravesical pressures of < or=40 cm-H₂O:a possible mechanism for pressure-induced cell proliferation[J]. World J Urol, 2008, 26(4):387-393.
20.	Kanematsu A, Ramachandran A, Adam RM. GATA-6 mediates human bladder smooth muscle differentiation:involvement of a novel enhancer element in regulating alpha-smooth muscle actin gene expression[J]. Am J Physiol Cell Physiol, 2007, 293(3):C1093-C1102.
21.	Nagata K, Sasaki E, Goda K, et al. Differences in heart rate variability in non-hypertensive diabetic patients correlate with the presence of underlying cerebrovascular disease[J]. Clin Physiol Funct Imaging, 2006, 26(2):92-98.
22.	Bouhout S, Gauvin R, Gibot L, et al. Bladder substitute reconstructed in a physiological pressure environment[J]. J Pediatr Urol, 2011, 7(3):276-282.
23.	Bouchelouche K, Horn T, Nordling J, et al. The action of cysteinyl-leukotrienes on intracellular calcium mobilization in human detrusor myocytes[J]. BJU Int, 2001, 87(7):690-696.

1. Chicurel ME, Chen CS, Ingber DE. Cellular control lies in the balance of forces[J]. Curr Opin Cell Biol, 1998, 10(2):232-239.
2. Galvin DJ, Watson RW, Gillespie JI, et al. Mechanical stretch regulates cell survival in human bladder smooth muscle cells in vitro[J]. Am J Physiol Renal Physiol, 2002, 283(6):1192-1199.
3. Orsola A, Adam RM, Peters CA, et al. The decision to undergo DNA or protein synthesis is determined by the degree of mechanical deformation in human bladder muscle cells[J]. Urology, 2002, 59(5):779-783.
4. Adam RM, Eaton SH, Estrada C, et al. Mechanical stretch is a highly selective regulator of gene expression in human bladder smooth muscle cells[J]. Physiol Genomics, 2004, 20(1):36-44.
5. Mirone V, Imbimbo C, Longo N, et al. The detrusor muscle:an innocent victim of bladder outlet obstruction[J]. Eur Urol, 2007, 51(1):57-66.
6. Shah MR, Wedgwood S, Czech L, et al. Cyclic stretch induces inducible nitric oxide synthase and soluble guanylate cyclase in pulmonary artery smooth muscle cells[J]. Int J Mol Sci, 2013, 14(2):4334-4348.
7. Lee SD, Akbal C, Jung C, et al. Intravesical pressure induces hyperplasia and hypertrophy of human bladder smooth muscle cells mediated by muscarinic receptors[J]. J Pediatr Urol, 2006, 2(4):271-276.
8. Vouyouka AG, Jiang Y, Rastogi R, et al. Ambient pressure upregulates nitric oxide synthase in a phosphorylated-extracellular regulated kinase-and protein kinase C-dependent manner[J]. J Vasc Surg, 2006, 44(5):1076-1084.
9. Wei X, Li DB, Xu F, et al. A novel bioreactor to simulate urinary bladder mechanical properties and compliance for bladder functional tissue engineering[J]. Chin Med J, 2011, 124(4):568-573.
10. Niklason LE, Gao J, Abbott WM, et al. Functional arteries grown in vitro[J]. Science, 1999, 284(5413):489-493.
11. Pörtner R, Nagel-Heyer S, Goepfert C, et al. Bioreactor design for tissue engineering[J]. J Biosci Bioeng, 2005, 100(3):235-245.
12. Farhat WA, Yeger H. Does mechanical stimulation have any role in urinary bladder tissue engineering?[J]. World J Urol, 2008, 26(4):301-305.
13. Butler DL, Goldstein S, Guilak F. Functional tissue engineering:the role of biomechanics[J]. J Biomech Eng, 2000, 122(6):570-575.
14. Kaspar D, Seidl W, Neidlinger-Wilke C, et al. Proliferation of human-derived osteoblast-like cells depends on the cycle number and frequency of uniaxial strain[J]. J Biomech, 2002, 35(7):873-880.
15. Leung DY, Glagov S, Mathews MB. Cyclic stretching stimulates synthesis of matrix components by arterial smooth muscle cells in vitro[J]. Science, 1976, 191(4226):475-477.
16. Houtchens GR, Foster MD, Desai TA, et al. Combined effects of microtopography and cyclic strain on vascular smooth muscle cell orientation[J]. J Biomech, 2008, 41(4):762-769.
17. Shah JB, Mcconkey DJ, Dinney CP. New strategies in muscle-invasive bladder cancer:on the road to personalized medicine[J]. Clin Cancer Res, 2011, 17(9):2608-2612.
18. Brown AL, Brook-Allred TT, Waddell JE, et al. Bladder acellular matrix as a substrate for studying in vitro bladder smooth muscle-urothelial cell interactions[J]. Biomaterials, 2005, 26(5):529-543.
19. Lee SD, Misseri R, Akbal C, et al. Muscarinic receptor expression increases following exposure to intravesical pressures of < or=40 cm-H₂O:a possible mechanism for pressure-induced cell proliferation[J]. World J Urol, 2008, 26(4):387-393.
20. Kanematsu A, Ramachandran A, Adam RM. GATA-6 mediates human bladder smooth muscle differentiation:involvement of a novel enhancer element in regulating alpha-smooth muscle actin gene expression[J]. Am J Physiol Cell Physiol, 2007, 293(3):C1093-C1102.
21. Nagata K, Sasaki E, Goda K, et al. Differences in heart rate variability in non-hypertensive diabetic patients correlate with the presence of underlying cerebrovascular disease[J]. Clin Physiol Funct Imaging, 2006, 26(2):92-98.
22. Bouhout S, Gauvin R, Gibot L, et al. Bladder substitute reconstructed in a physiological pressure environment[J]. J Pediatr Urol, 2011, 7(3):276-282.
23. Bouchelouche K, Horn T, Nordling J, et al. The action of cysteinyl-leukotrienes on intracellular calcium mobilization in human detrusor myocytes[J]. BJU Int, 2001, 87(7):690-696.

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