• 四川大學(xué)華西醫(yī)院（成都，610041）1 泌尿外科，2 移植免疫重點(diǎn)實(shí)驗(yàn)室;

目的 研究靜水壓作用下人膀胱平滑肌細(xì)胞（human bladder smooth muscle cells，hb-SMCs）內(nèi)游離鈣離
子濃度(intracellular free calcium concentration，[Ca2+]i) 的變化及香草型瞬時(shí)受體電位（transient receptor potential vanilloid，TRPV）基因的表達(dá)情況，初步探討機(jī)械應(yīng)力在介導(dǎo)hb-SMCs 增殖中的分子機(jī)制。 方法 取第6 ～ 7 代細(xì)胞進(jìn)行實(shí)驗(yàn)，采用新型Ca2+ 熒光染色劑Fluo-3/AM 負(fù)載hb-SMCs，應(yīng)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(nèi)[Ca2+]i。采用RT-PCR 技術(shù)測定200 cm H2O 靜水壓作用0、2、6、12、24 h 時(shí)TRPV1、TRPV2 及TRPV4 基因mRNA 的表達(dá)量。 結(jié)果 A、B、C 組hb-SMCs 內(nèi)[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(dá)，隨時(shí)間推移表達(dá)無明顯變化；各組間TRPV1、TRPV2、TRPV4 mRNA 表達(dá)量比較差異均無統(tǒng)計(jì)學(xué)意義（P  gt; 0.05）。 結(jié)論 高水平靜水壓作用下hb-SMCs 內(nèi)[Ca2+]i 明顯升高。作為可能表達(dá)為機(jī)械敏感性陽離子通道的TRPV1、TRPV2 及TRPV4 基因在hb-SMCs 中均有表達(dá)，機(jī)械應(yīng)力可能通過調(diào)節(jié)其開放量而非上調(diào)其表達(dá)量使hb-SMCs 內(nèi)[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(duì)人膀胱平滑肌細(xì)胞內(nèi)鈣離子濃度及香草型瞬時(shí)受體電位表達(dá)的影響. 中國修復(fù)重建外科雜志, 2012, 26(4): 457-460. doi: 復(fù)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(duì)細(xì)胞施加力學(xué)作用的實(shí)驗(yàn)方法. 航天醫(yī)學(xué)與醫(yī)學(xué)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(yīng)力施加方式對(duì)膀胱平滑肌細(xì)胞影響的比較研究. 四川大學(xué)學(xué)報(bào): 醫(yī)學(xué)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. 1. Atala A, Bauer SB, Soker S, et al. Tissue-engineered autologous bladders for patients needing cystoplasty. Lancet, 2006, 367(9518): 1241-1246.
  2. 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. 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. 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. 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. 6. Landsberg JW, Yuan JX. Calcium and TRP channels in pulmonary vascular smooth muscle cell proliferation. News Physiol Sci, 2004, 19: 44-50.
  7. 7. O’Neil RG, Heller S. The mechanosensitive nature of TRPV channels. Pflugers Arch, 2005, 451(1): 193-203.
  8. 8. 曾燁, 劉肖珩. 對(duì)細(xì)胞施加力學(xué)作用的實(shí)驗(yàn)方法. 航天醫(yī)學(xué)與醫(yī)學(xué)工程, 2007, 20(3): 227-234.
  9. 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. 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. 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. 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. 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. 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. 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. 16. Christensen AP, Corey DP. TRP channels in mechanosensation: direct or indirect activation? Nat Rev Neurosci, 2007, 8(7): 510-521.
  17. 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. 18. Araki I. TRP channels in urinary bladder mechanosensation. Adv Exp Med Biol, 2011, 704: 861-879.
  19. 19. Hardie RC. A brief history of trp: commentary and personal perspective. Pflugers Arch, 2011, 461(5): 493-498.
  20. 20. 鄔韜, 陳林, 王彥, 等. 不同應(yīng)力施加方式對(duì)膀胱平滑肌細(xì)胞影響的比較研究. 四川大學(xué)學(xué)報(bào): 醫(yī)學(xué)版, 2011, 42(6): 846-849.
  21. 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.