靜水壓對人膀胱平滑肌細胞內鈣離子濃度及香草型瞬時受體電位表達的影響_《中國修復重建外科雜志》

作者：

韓振偉 ¹ , 王坤杰 ¹ , 陳林 ¹ , 韋堂墻 ¹ , 羅德毅 ¹ , 李勝富 ²

四川大學華西醫院（成都，610041）1 泌尿外科，2 移植免疫重點實驗室;

關鍵詞：

靜水壓人膀胱平滑肌細胞游離鈣離子濃度香草型瞬時受體電位機械敏感性陽離子通道

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目的研究靜水壓作用下人膀胱平滑肌細胞（human bladder smooth muscle cells，hb-SMCs）內游離鈣離
子濃度(intracellular free calcium concentration，[Ca2+]i) 的變化及香草型瞬時受體電位（transient receptor potential vanilloid，TRPV）基因的表達情況，初步探討機械應力在介導hb-SMCs 增殖中的分子機制。方法取第6 ～ 7 代細胞進行實驗，采用新型Ca2+ 熒光染色劑Fluo-3/AM 負載hb-SMCs，應用倒置激光掃描共聚焦顯微鏡分別測定靜水壓加壓0 cm H2O
（1 cm H2O=0.098 kPa）（A 組）、200 cm H2O 作用30 min（B 組）及撤銷200 cm H2O 靜水壓（C 組）后hb-SMCs 內[Ca2+]i。采用RT-PCR 技術測定200 cm H2O 靜水壓作用0、2、6、12、24 h 時TRPV1、TRPV2 及TRPV4 基因mRNA 的表達量。結果 A、B、C 組hb-SMCs 內[Ca2+]i 分別為（100.808 ± 1.724）、（122.008 ± 1.575）、（99.918 ± 0.887）U，B 組[Ca2+]i 明顯高于A、C 組（P lt; 0.001），C 組撤銷200 cm H2O 靜水壓力后[Ca2+]i 下降至A 組基線水平（t=0.919，P=0.394）。RT-PCR 檢測示，200 cm H2O 靜水壓加壓0 h 組、2 h 組、6 h 組、12 h 組及24 h 組TRPV1、TRPV2 及TRPV4 在hb-SMCs 中均有表達，隨時間推移表達無明顯變化；各組間TRPV1、TRPV2、TRPV4 mRNA 表達量比較差異均無統計學意義（P gt; 0.05）。結論高水平靜水壓作用下hb-SMCs 內[Ca2+]i 明顯升高。作為可能表達為機械敏感性陽離子通道的TRPV1、TRPV2 及TRPV4 基因在hb-SMCs 中均有表達，機械應力可能通過調節其開放量而非上調其表達量使hb-SMCs 內[Ca2+]i 升高。

引用本文： 韓振偉 ,王坤杰,陳林,韋堂墻,羅德毅,李勝富. 靜水壓對人膀胱平滑肌細胞內鈣離子濃度及香草型瞬時受體電位表達的影響. 中國修復重建外科雜志, 2012, 26(4): 457-460. doi: 復制

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.	曾燁, 劉肖珩. 對細胞施加力學作用的實驗方法. 航天醫學與醫學工程, 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.	鄔韜, 陳林, 王彥, 等. 不同應力施加方式對膀胱平滑肌細胞影響的比較研究. 四川大學學報: 醫學版, 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. 曾燁, 劉肖珩. 對細胞施加力學作用的實驗方法. 航天醫學與醫學工程, 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. 鄔韜, 陳林, 王彥, 等. 不同應力施加方式對膀胱平滑肌細胞影響的比較研究. 四川大學學報: 醫學版, 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.

《中國修復重建外科雜志》

靜水壓對人膀胱平滑肌細胞內鈣離子濃度及香草型瞬時受體電位表達的影響

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《中國修復重建外科雜志》

靜水壓對人膀胱平滑肌細胞內鈣離子濃度及香草型瞬時受體電位表達的影響

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Content

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