压力性尿失禁大鼠中环氧合酶2表达的变化及意义
|
摘要
目的压力性尿失禁(SUI)的发病机制研究较少。文中旨在检测环氧合酶2(COX?2)在SUI大鼠模型中的尿道、阴道、尿道平滑肌中的表达变化。方法健康雌性未育SD大鼠60只,雄性SD大鼠15只,合拢自然妊娠,取分娩后雌性大鼠分别行扩张阴道(单纯扩张组)、扩张阴道+切除卵巢(扩张+切除组)建立SUI大鼠模型,另使用正常妊娠分娩后大鼠为对照组,每组6只。通过喷嚏实验及尿动力学检查最大膀胱容量(MBC)和腹压漏尿点压(ALPP),荧光定量PCR、Western blot检测COX?2 mRNA及蛋白水平的表达,免疫组化检测COX?2在尿道、阴道、尿道平滑肌的表达。结果扩张+切除组、单纯扩张组ALPP较对照组明显下降,扩张+切除组较单纯扩张组明显下降(P<0.05)。单纯扩张组、扩张+切除组COX?2mRNA表达量[(1.40±0.07)、(1.75±0.10)]、蛋白的相对表达量[(1.64±0.05)、(1.95±0.08)]较对照组[(1.00±0.16)、(0.77±0.26)]均明显升高(P<0.001);扩张+切除组较单纯扩张组明显升高(P<0.05)。免疫组化结果显示,对照组、单纯扩张组、扩张+切除组阴道组织中COX2的染色强度表达积分依次为0.50±0.54、5.55±0.54、9.33±0.81,组间两两比较差异有统计学意义(P<0.05);尿道平滑肌组织中染色强度表达依次为0.66±0.51、5.33±0.51、8.50±0.54,组间两两比较差异有统计学差异(P<0.05)。结论切除卵巢后的雌激素下降可致COX?2表达的升高,COX?2与SUI的发病机制相关。
Objective There are a few researches on the mechanism of stress urinary incontinence(SUI). The article aimed to examine the changes of COX-2 expression in the urethra,vagina and urethral smooth muscle of SUI rat mode to evaluate the effect of estro?gen on COX-2 expression.Methods Sixty unbearing healthy fe?male SD rats and fifteen male SD rats were gathered for spontaneous delivery. SUI rat models were constructed using expanded vagina,ex?panded vagina + ovariectomy respectively after delivery,which were expanded vagina group and expanded vagina + ovariectomy group.Six successfully modeled rats were chosen for the follow-up experiment. SD rats modeled after normal pregnancy were the control group. Sneezing experiment and urodynamic examination were used to examine the maximum bladder capacity(MBC)and abdominal leak point pressure(ALPP). Fluorescent quantitative PCR and western blot were applied to detect the expressions of COX-2 mRNA and protein,and immunohistochemistry was used to detect the expressions of COX-2 in urethra,vagina and urethral smooth muscle.Results Compared with control group,ALPP in two experimental groups were significantly decreased,among which ALPP in ex?panded vagina + ovariectomy group was significantly decreased in comparison to expanded vagina group(P<0.05). Compared with con?trol group,the expressions of COX-2 mRNA and protein in expanded vagina group and expanded vagina+ovariectomy group were sig?nificantly higher,among which the figures in expanded vagina+ovariectomy group were significantly higher than those in expanded va?gina group(P<0.05). The result of immunohistochemistry showed staining intensity integral expression of COX-2 in vaginal tissues of control group,expanded vagina group and expanded vagina+ovariectomy group were 0.50±0.54,5.55±0.54,9.33±0.81,so differenc?es between any two groups were of statistical significance(P<0.05);staining intensity integral expression of COX-2 in urethral smooth muscle of control group,expanded vagina group and expanded vagina+ovariectomy group were 0.66±0.51,5.33±0.51,8.50±0.54,so differences between any two groups were of statistical significance(P<0.05).Conclusion The expression of COX-2 was related to the mechanism of SUI. The decrease of estrogen may increase the expression of COX-2 in SUI rats,which supports the treatment of SUI.
Objective There are a few researches on the mechanism of stress urinary incontinence(SUI). The article aimed to examine the changes of COX-2 expression in the urethra,vagina and urethral smooth muscle of SUI rat mode to evaluate the effect of estro?gen on COX-2 expression.Methods Sixty unbearing healthy fe?male SD rats and fifteen male SD rats were gathered for spontaneous delivery. SUI rat models were constructed using expanded vagina,ex?panded vagina + ovariectomy respectively after delivery,which were expanded vagina group and expanded vagina + ovariectomy group.Six successfully modeled rats were chosen for the follow-up experiment. SD rats modeled after normal pregnancy were the control group. Sneezing experiment and urodynamic examination were used to examine the maximum bladder capacity(MBC)and abdominal leak point pressure(ALPP). Fluorescent quantitative PCR and western blot were applied to detect the expressions of COX-2 mRNA and protein,and immunohistochemistry was used to detect the expressions of COX-2 in urethra,vagina and urethral smooth muscle.Results Compared with control group,ALPP in two experimental groups were significantly decreased,among which ALPP in ex?panded vagina + ovariectomy group was significantly decreased in comparison to expanded vagina group(P<0.05). Compared with con?trol group,the expressions of COX-2 mRNA and protein in expanded vagina group and expanded vagina+ovariectomy group were sig?nificantly higher,among which the figures in expanded vagina+ovariectomy group were significantly higher than those in expanded va?gina group(P<0.05). The result of immunohistochemistry showed staining intensity integral expression of COX-2 in vaginal tissues of control group,expanded vagina group and expanded vagina+ovariectomy group were 0.50±0.54,5.55±0.54,9.33±0.81,so differenc?es between any two groups were of statistical significance(P<0.05);staining intensity integral expression of COX-2 in urethral smooth muscle of control group,expanded vagina group and expanded vagina+ovariectomy group were 0.66±0.51,5.33±0.51,8.50±0.54,so differences between any two groups were of statistical significance(P<0.05).Conclusion The expression of COX-2 was related to the mechanism of SUI. The decrease of estrogen may increase the expression of COX-2 in SUI rats,which supports the treatment of SUI.
引文
[1]Irwin DE,Milsom I,Hunskaar S,et al.Population-Based Survey of Urinary Incontinence,Overactive Bladder,and Other Lower Urinary Tract Symptoms in Five Countries:Results of the EPIC Study[J].EurUrol,2006,50(6):1306-1315.
[2]田丽,古丽娜·阿巴拜克力;布海切木·卡德尔.乌鲁木齐市部分社区成年女性压力性尿失禁的流行病学及危险因素分析[J].医学研究生学报,2012,25(7):713-718.
[3]卞飞,周峰,陈卫国,等.尿道中段悬吊术治疗压力性尿失禁的疗效及安全性分析[J].医学研究生学报,2015,28(6):618-621.
[4]Lin G,Shindel AW,Banie L,et al.Molecular Mechanisms Related to Parturition-Induced Stress Urinary Incontinence[J].EurUrol,2009,55(5):1213-1223.
[5]Lenis AT,Kuang M,Woo LL,et al.Impact of Parturition on Chemokine Homing Factor Expression in the Vaginal Distention Model of Stress Urinary Incontinence[J].J Urol,2013,189(4):1588-1594.
[6]Marentette JO,Hurst RE,McHowat J.Impaired Expression of Prostaglandin E2(PGE2)Synthesis and Degradation Enzymes during Differentiation of Immortalized Urothelial Cells from Patients with Interstitial Cystitis/Painful Bladder Syndrome[J].Plos One,2015,10(6):e129466.
[7]Giannantoni A.Editorial comment on:Intravesicalbotulinum toxin Aadministration inhibits COX-2 and EP4 expression and suppresses bladderhyperactivity in cyclophosphamide-induced cystitis in rats[J].EurUrol,2009,56(1):166-167.
[8]杨智明,方克伟.诱导型一氧化氮合酶/环氧合酶2在女性压力性尿失禁中的研究进展[J].医学研究生学报,2015,28(3):323-328.
[9]Lin YH,Liu G,Li M,et al.Recovery of continence function following simulated birth trauma involves repair of muscle and nerves in the urethra in the female mouse[J].Eur Urol,2010,57(3):506-512.
[10]Dokmeci F,Seval M,Gok H.Comparison of ambulatory versus conventional urodynamics in females with urinary incontinence[J].Neurourol Urodyn,2010,29(4):518-521.
[11]de Jongh R,van Koeveringe GA,van Kerrebroeck PE,Markerink-van Ittersum M,de Vente J,Gillespie JI.The effects of exogenous prostaglandins and the identification of constitutive cyclooxygenase I and II immunoreactivity in the normal guinea pig bladder[J].BJU Int,2007,100(2):419-429.
[12]Altman D,Ekstr?m A,Forsgren C,et al.Symptoms of anal and urinary incontinence following cesarean section or spontaneous vaginal delivery[J].Am J Obstet Gynecol,2007,197(5):512.e1-7.
[13]Morita I,Schindler M,Regier MK,et al.Different intracellular locations for prostaglandin endoperoxide H synthase-1 and-2[J].J Biol Chem,1995,270(18):10902-10908.
[14]Khan MA,Thompson CS,Mumtaz FH,et al.Role of prostaglandins in the urinary bladder:an update[J].Prostaglandins Leukot Essent Fatty Acids,1998,59(6):415-422.
[15]Comeglio P,Chavalmane AK,Fibbi B,et al.Human prostatic urethra expresses vitamin D receptor and responds to vitamin Dreceptor ligation[J].J Endocrinol Invest,2010,33(10):730-738.
[16]Yamamichi F,Shigemura K,Behnsawy HM,et al.Beta-3 adrenergic receptors could be significant factors for overactive bladder-related symptoms[J].Int J Clin Exp Pathol,2015,8(9):11863-11870.
[17]Hashitani H.Interaction between interstitial cells and smooth muscles in the lower urinary tract and penis[J].J Physiol,2010,576(3):707-714.
[18]Murakami M,Nakashima K,Kamei D,et al.Cellular prostaglandin E2 production by membrane-boundprostaglandin E synthase-2 via both cyclooxygenases-1 and-2[J].J Biol Chem,2003,278(39):37937-37947.
[19]Henderson C,Bainbridge J,Keaton K,et al.Synergistic effect of vaginal trauma and ovariectomy in a murine model of stress urinary incontinence:upregulation of urethral nitric oxide synthases and estrogen receptors[J].Mediators Inflamm,2014:314846.
[2]田丽,古丽娜·阿巴拜克力;布海切木·卡德尔.乌鲁木齐市部分社区成年女性压力性尿失禁的流行病学及危险因素分析[J].医学研究生学报,2012,25(7):713-718.
[3]卞飞,周峰,陈卫国,等.尿道中段悬吊术治疗压力性尿失禁的疗效及安全性分析[J].医学研究生学报,2015,28(6):618-621.
[4]Lin G,Shindel AW,Banie L,et al.Molecular Mechanisms Related to Parturition-Induced Stress Urinary Incontinence[J].EurUrol,2009,55(5):1213-1223.
[5]Lenis AT,Kuang M,Woo LL,et al.Impact of Parturition on Chemokine Homing Factor Expression in the Vaginal Distention Model of Stress Urinary Incontinence[J].J Urol,2013,189(4):1588-1594.
[6]Marentette JO,Hurst RE,McHowat J.Impaired Expression of Prostaglandin E2(PGE2)Synthesis and Degradation Enzymes during Differentiation of Immortalized Urothelial Cells from Patients with Interstitial Cystitis/Painful Bladder Syndrome[J].Plos One,2015,10(6):e129466.
[7]Giannantoni A.Editorial comment on:Intravesicalbotulinum toxin Aadministration inhibits COX-2 and EP4 expression and suppresses bladderhyperactivity in cyclophosphamide-induced cystitis in rats[J].EurUrol,2009,56(1):166-167.
[8]杨智明,方克伟.诱导型一氧化氮合酶/环氧合酶2在女性压力性尿失禁中的研究进展[J].医学研究生学报,2015,28(3):323-328.
[9]Lin YH,Liu G,Li M,et al.Recovery of continence function following simulated birth trauma involves repair of muscle and nerves in the urethra in the female mouse[J].Eur Urol,2010,57(3):506-512.
[10]Dokmeci F,Seval M,Gok H.Comparison of ambulatory versus conventional urodynamics in females with urinary incontinence[J].Neurourol Urodyn,2010,29(4):518-521.
[11]de Jongh R,van Koeveringe GA,van Kerrebroeck PE,Markerink-van Ittersum M,de Vente J,Gillespie JI.The effects of exogenous prostaglandins and the identification of constitutive cyclooxygenase I and II immunoreactivity in the normal guinea pig bladder[J].BJU Int,2007,100(2):419-429.
[12]Altman D,Ekstr?m A,Forsgren C,et al.Symptoms of anal and urinary incontinence following cesarean section or spontaneous vaginal delivery[J].Am J Obstet Gynecol,2007,197(5):512.e1-7.
[13]Morita I,Schindler M,Regier MK,et al.Different intracellular locations for prostaglandin endoperoxide H synthase-1 and-2[J].J Biol Chem,1995,270(18):10902-10908.
[14]Khan MA,Thompson CS,Mumtaz FH,et al.Role of prostaglandins in the urinary bladder:an update[J].Prostaglandins Leukot Essent Fatty Acids,1998,59(6):415-422.
[15]Comeglio P,Chavalmane AK,Fibbi B,et al.Human prostatic urethra expresses vitamin D receptor and responds to vitamin Dreceptor ligation[J].J Endocrinol Invest,2010,33(10):730-738.
[16]Yamamichi F,Shigemura K,Behnsawy HM,et al.Beta-3 adrenergic receptors could be significant factors for overactive bladder-related symptoms[J].Int J Clin Exp Pathol,2015,8(9):11863-11870.
[17]Hashitani H.Interaction between interstitial cells and smooth muscles in the lower urinary tract and penis[J].J Physiol,2010,576(3):707-714.
[18]Murakami M,Nakashima K,Kamei D,et al.Cellular prostaglandin E2 production by membrane-boundprostaglandin E synthase-2 via both cyclooxygenases-1 and-2[J].J Biol Chem,2003,278(39):37937-37947.
[19]Henderson C,Bainbridge J,Keaton K,et al.Synergistic effect of vaginal trauma and ovariectomy in a murine model of stress urinary incontinence:upregulation of urethral nitric oxide synthases and estrogen receptors[J].Mediators Inflamm,2014:314846.