EFFECT OF HYDROSTATIC PRESSURE ON INTRACELLULAR FREE CALCIUM CONCENTRATION AND TRANSIENT RECEPTOR POTENTIAL VANILLOID EXPRESSION IN HUMAN BLADDER SMOOTH MUSCLE CELLS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

HAN Zhenwei ¹ , WANG Kunjie ¹ , CHEN Lin ¹ , WEI Tangqiang ¹ , LUO Deyi ¹ , LI Shengfu. ²

1Department of Urology, 2Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China.;

Corresponding author：

Keywords：

Hydrostatic pressure; Human bladder smooth muscle cells; Intracellular free calcium concentration; Transient receptor potential vanilloid; Stretch-activated cation channel

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Objective To explore the effect of hydrostatic pressure on intracellular free calcium concentration ([Ca2+]i) and the gene expression of transient receptor potential vanilloid (TRPV) in cultured human bladder smooth muscle cells (hb-SMCs), and to prel iminarily probe into the possible molecular mechanism of hb-SMCs prol iferation stimulated by hydrostatic pressure. Methods The passage 6-7 hb-SMCs were loaded with Ca2+ indicator Fluo-3/AM. When the hb-SMCs were under 0 cm H2O (1 cm H2O=0.098 kPa) (group A) or 200 cm H2O hydrostatic pressure for 30 minutes (group B) and then removing the 200 cm H2O hydrostatic pressure (group C), the [Ca2+]i was measured respectively by inverted laser anning
confocal microscope. When the hb-SMCs were given the 200 cm H2O hydrostatic pressure for 0 hour, 2 hours, 6 hours, 2 hours, and 24 hours, the mRNA expressions of TRPV1, TRPV2, and TRPV4 were detected by RT-PCR technique. Results The [Ca2+]i of group A, group B, and group C were (100.808 ± 1.724), (122.008 ± 1.575), and (99.918 ± 0.887) U, respectively; group B was significantly higher than groups A and C (P lt; 0.001). The [Ca2+]i of group C decreased to the base l ine level of group A after removing the pressure (t=0.919, P=0.394). The TRPV1, TRPV2, and TRPV4 genes expressed in hb-SMCs under 200 cm H2O hydrostatic pressure at 0 hour, 2 hours, 6 hours, 12 hours, and 24 hours, but the expressions had no obvious changes with time. There was no significant difference in the expressions of TRPV1, TRPV2, and TRPV4 among 3 groups (P gt; 0.05). Conclusion The [Ca2+]i of hb-SMCs increases significantly under high hydrostatic pressure. As possible genes in stretch-activated cation channel, the TRPV1, TRPV2, and TRPV4 express in hb-SMCs under 200 cm H2O hydrostatic pressure. It is possible that the mechanical pressure regulates the [Ca2+]i of hb-SMCs by opening the stretch-activated cation channel rather than up-regulating its expression.

Citation： HAN Zhenwei,WANG Kunjie,CHEN Lin,WEI Tangqiang,LUO Deyi,LI Shengfu.. EFFECT OF HYDROSTATIC PRESSURE ON INTRACELLULAR FREE CALCIUM CONCENTRATION AND TRANSIENT RECEPTOR POTENTIAL VANILLOID EXPRESSION IN HUMAN BLADDER SMOOTH MUSCLE CELLS. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(4): 457-460. doi: Copy

1.	Atala A, Bauer SB, Soker S, et al. Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet, 2006, 367(9518): 1241-1246.
2.	Li YS, Haga JH, Chien S. Molecular basis of the effects of shear stress on vascular endothelial cells. J Biomech, 2005, 38(10): 1949-1971.
3.	Koyama Y, Mitsui N, Suzuki N, et al. Effect of compressive force on the expression of inflammatory cytokines and their receptors in osteoblastic Saos-2 cells. Arch Oral Biol, 2008, 53(5): 488-496.
4.	Ito S, Kume H, Naruse K, et al. A novel Ca2+ influx pathway activated by mechanical stretch in human airway smooth muscle cells. Am J Respir Cell Mol Biol, 2008, 38(4): 407-413.
5.	Guibert C, Ducret T, Savineau JP. Expression and physiological roles of TRP channels in smooth muscle cells. Adv Exp Med Biol, 2011, 704: 687-706.
6.	Landsberg JW, Yuan JX. Calcium and TRP channels in pulmonary vascular smooth muscle cell proliferation. News Physiol Sci, 2004, 19: 44-50.
7.	O’Neil RG, Heller S. The mechanosensitive nature of TRPV channels. Pflugers Arch, 2005, 451(1): 193-203.
8.	曾烨, 刘肖珩. 对细胞施加力学作用的实验方法. 航天医学与医学工程, 2007, 20(3): 227-234.
9.	Zhang S, Yuan JX, Barrett KE, et al. Role of Na+/Ca2+ exchange in regulating cytosolic Ca2+ in cultured human pulmonary artery smooth muscle cells. Am J Physiol Cell Physiol, 2005, 288(2): C245-252.
10.	Kumar A, Knox AJ, Boriek AM. CCAAT/enhancer-binding protein and activator protein-1 transcription factors regulate the expression of interleukin-8 through the mitogen-activated protein kinase pathways in response to mechanical stretch of human airway smooth muscle cells. J Biol Chem, 2003, 278(21): 18868-18876.
11.	Golovina VA, Platoshyn O, Bailey CL, et al. Upregulated TRP and enhanced capacitative Ca2+ entry in human pulmonary artery myocytes during proliferation. Am J Physiol Heart Circ Physiol, 2001, 280(2): H746-755.
12.	Tolekova AN, Hadzhibozheva PV, Iliev RN, et al. Participation of extracellular Ca (2+) or ghrelin in peptide-mediated contraction of strips from rat urinary bladder. Regul Pept, 2010, 162(1-3): 79-83.
13.	Dalrymple A, Mahn K, Poston L, et al. Mechanical stretch regulates TRPC expression and calcium entry in human myometrial smooth muscle cells. Mol Hum Reprod, 2007, 13(3): 171-179.
14.	Park KS, Kim Y, Lee YH, et al. Mechanosensitive cation channels in arterial smooth muscle cells are activated by diacylglycerol and inhibited by phospholipase C inhibitor. Circ Res, 2003, 93(6): 557-564.
15.	Beech DJ, Muraki K, Flemming R. Non-selective cationic channels of smooth muscle and the mammalian homologues of Drosophila TRP. J Physiol, 2004, 559(Pt 3): 685-706.
16.	Christensen AP, Corey DP. TRP channels in mechanosensation: direct or indirect activation? Nat Rev Neurosci, 2007, 8(7): 510-521.
17.	Lazzeri M, Costantini E, Porena M. TRP family proteins in the lower urinary tract: translating basic science into new clinical prospective. Ther Adv Urol, 2009, 1(1): 33-42.
18.	Araki I. TRP channels in urinary bladder mechanosensation. Adv Exp Med Biol, 2011, 704: 861-879.
19.	Hardie RC. A brief history of trp: commentary and personal perspective. Pflugers Arch, 2011, 461(5): 493-498.
20.	邬韬, 陈林, 王彦, 等. 不同应力施加方式对膀胱平滑肌细胞影响的比较研究. 四川大学学报: 医学版, 2011, 42(6): 846-849.
21.	Yu Y, Fantozzi I, Remillard CV, et al. Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension. Proc Natl Acad Sci U S A, 2004, 101(38): 13861-13866.

1. Atala A, Bauer SB, Soker S, et al. Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet, 2006, 367(9518): 1241-1246.
2. Li YS, Haga JH, Chien S. Molecular basis of the effects of shear stress on vascular endothelial cells. J Biomech, 2005, 38(10): 1949-1971.
3. Koyama Y, Mitsui N, Suzuki N, et al. Effect of compressive force on the expression of inflammatory cytokines and their receptors in osteoblastic Saos-2 cells. Arch Oral Biol, 2008, 53(5): 488-496.
4. Ito S, Kume H, Naruse K, et al. A novel Ca2+ influx pathway activated by mechanical stretch in human airway smooth muscle cells. Am J Respir Cell Mol Biol, 2008, 38(4): 407-413.
5. Guibert C, Ducret T, Savineau JP. Expression and physiological roles of TRP channels in smooth muscle cells. Adv Exp Med Biol, 2011, 704: 687-706.
6. Landsberg JW, Yuan JX. Calcium and TRP channels in pulmonary vascular smooth muscle cell proliferation. News Physiol Sci, 2004, 19: 44-50.
7. O’Neil RG, Heller S. The mechanosensitive nature of TRPV channels. Pflugers Arch, 2005, 451(1): 193-203.
8. 曾烨, 刘肖珩. 对细胞施加力学作用的实验方法. 航天医学与医学工程, 2007, 20(3): 227-234.
9. Zhang S, Yuan JX, Barrett KE, et al. Role of Na+/Ca2+ exchange in regulating cytosolic Ca2+ in cultured human pulmonary artery smooth muscle cells. Am J Physiol Cell Physiol, 2005, 288(2): C245-252.
10. Kumar A, Knox AJ, Boriek AM. CCAAT/enhancer-binding protein and activator protein-1 transcription factors regulate the expression of interleukin-8 through the mitogen-activated protein kinase pathways in response to mechanical stretch of human airway smooth muscle cells. J Biol Chem, 2003, 278(21): 18868-18876.
11. Golovina VA, Platoshyn O, Bailey CL, et al. Upregulated TRP and enhanced capacitative Ca2+ entry in human pulmonary artery myocytes during proliferation. Am J Physiol Heart Circ Physiol, 2001, 280(2): H746-755.
12. Tolekova AN, Hadzhibozheva PV, Iliev RN, et al. Participation of extracellular Ca (2+) or ghrelin in peptide-mediated contraction of strips from rat urinary bladder. Regul Pept, 2010, 162(1-3): 79-83.
13. Dalrymple A, Mahn K, Poston L, et al. Mechanical stretch regulates TRPC expression and calcium entry in human myometrial smooth muscle cells. Mol Hum Reprod, 2007, 13(3): 171-179.
14. Park KS, Kim Y, Lee YH, et al. Mechanosensitive cation channels in arterial smooth muscle cells are activated by diacylglycerol and inhibited by phospholipase C inhibitor. Circ Res, 2003, 93(6): 557-564.
15. Beech DJ, Muraki K, Flemming R. Non-selective cationic channels of smooth muscle and the mammalian homologues of Drosophila TRP. J Physiol, 2004, 559(Pt 3): 685-706.
16. Christensen AP, Corey DP. TRP channels in mechanosensation: direct or indirect activation? Nat Rev Neurosci, 2007, 8(7): 510-521.
17. Lazzeri M, Costantini E, Porena M. TRP family proteins in the lower urinary tract: translating basic science into new clinical prospective. Ther Adv Urol, 2009, 1(1): 33-42.
18. Araki I. TRP channels in urinary bladder mechanosensation. Adv Exp Med Biol, 2011, 704: 861-879.
19. Hardie RC. A brief history of trp: commentary and personal perspective. Pflugers Arch, 2011, 461(5): 493-498.
20. 邬韬, 陈林, 王彦, 等. 不同应力施加方式对膀胱平滑肌细胞影响的比较研究. 四川大学学报: 医学版, 2011, 42(6): 846-849.
21. Yu Y, Fantozzi I, Remillard CV, et al. Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension. Proc Natl Acad Sci U S A, 2004, 101(38): 13861-13866.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF HYDROSTATIC PRESSURE ON INTRACELLULAR FREE CALCIUM CONCENTRATION AND TRANSIENT RECEPTOR POTENTIAL VANILLOID EXPRESSION IN HUMAN BLADDER SMOOTH MUSCLE CELLS

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