胰岛素样生长因子-Ⅰ对雌性大鼠尿道括约肌损伤修复的影响及其机制研究
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摘要
目的:探讨经尿道置管的尿动力学检测方法在大鼠压力性尿失禁模型尿动力学研究中应用的可行性和有效性,并分析大鼠尿道括约肌功能障碍的尿动力学特征。
方法:12只成年雌性未育大鼠在模拟产伤致尿道括约肌损伤前及损伤后第14天经尿道置管行充盈期膀胱测压、漏尿点压力和静态尿道压力测定。
结果10只顺利完成实验,损伤前平均最大膀胱容量、排尿压、腹压漏尿点压、最大尿道压、最大尿道闭合压分别为(0.88±0.46)ml、(26.50±6.42)cmH_2O (1cmH_2O=0.098KPa)、(25.65±5.47)cmH_2O、(14.80±2.78)cmH_2O和(13.10±2.60)cmH_2O,损伤后分别为(1.47±0.69)ml、(17.10±6.74)cmH_2O、(15.55±5.24)cmH_2O、(12.10±3.07)cmH_2O和(10.60±3.20)cmH_2O,大鼠最大膀胱容量增加(0.60±0.40)ml(P<0.01),排尿压、腹压漏尿点压分别下降(9.40±8.74)cmH_2O、(9.50±4.92)cmH_2O(P<0.05),最大尿道压和最大尿道闭合压亦下降(2.70±3.37)cmH_2O、(2.50±2.59)cmH_2O(P<0.05)。
结论模拟产伤可有效建立雌性大鼠压力性尿失禁模型,经尿道置管的尿动力检测方法能客观评价大鼠尿道括约肌功能。
目的:观察雌性Sprague - Dawley(SD)大鼠尿道括约肌损伤后组织修复过程中局部内源性胰岛素样生长因子-I和II的mRNA表达情况。研究胰岛素样生长因子-I局部注射在其损伤后修复过程中对内源性IGF-I和II mRNA表达的影响及对尿道显微形态结构改变的影响。
方法:制作模拟产伤的SD大鼠尿道括约肌损伤模型后,①分别于损伤后2、4、6、8、14天处死大鼠取出尿道,行逆转录-聚合酶链反应(RT- PCR),琼脂糖凝胶电泳,通过辉度扫描与内参照对比,了解IGF– I和II的mRNA表达变化;②于尿道中段局部注射IGF-I,RT-PCR半定量分析损伤后2、4、6、8、14d时IGF-I和II mRNA表达;③于尿道中段局部注射IGF-I,中段尿道Masson’s trichrome染色,观察其显微结构动态改变,并运用计算机图象分析系统定量分析尿道横纹肌、平滑肌及结缔组织组成。尿道括约肌无损伤大鼠为正常对照组。
结果:①IGF - I mRNA在损伤后4、6、8、14天表达,尤以第8天升高明显(P<0.05);IGF-II mRNA在伤后4、6、8天表达,以第6、8天升高明显(P<0.05);未损伤组无明显IGF表达。②试验组内源性IGF-I mRNA在伤后2、4、6、8、14d均有表达,尤以第8d增加明显,与生理盐水对照组比较于第4、14d表达有极显著增加(P<0.01),第8d有显著增加(P<0.05);试验组IGF-II mRNA在伤后4、6、8、14d表达,以第6、8d增加明显,与生理盐水对照组比较第4d有极显著增加(P<0.01)。③损伤后尿道显微结构发生明显改变,尿道横纹肌、平滑肌出现断裂,萎缩,排列结构紊乱,胶原结缔组织增生明显。试验组与生理盐水对照组比较于8d、14d肌组织所占尿道横截面积比例明显增高(P<0.05),肌纤维增生丰富,形态结构更为完整。
结论:SD大鼠尿道括约肌损伤修复过程中有内源性IGF-I和II的mRNA表达变化,提示IGF参与尿道括约肌损伤后肌细胞再生修复过程;局部注射IGF-I能促进尿道括约肌损伤局部内源性IGF-I和II mRNA的表达,并能促进尿道括约肌肌细胞损伤后的修复,使尿道括约肌形态结构更为完整,提示IGF-I对尿道括约肌损伤及其所致压力性尿失禁具有潜在的治疗作用。
Objective: To evaluate the effect of a new method of urodynamics in the research of female rat stress urinary incontinence model and present the urodynamic characteristic of urethral sphincter dysfunction of rat.
Method: Twelve female virgin SD rats were performed urethral sphincter muscle injury by simulating a birth trauma. They all underwent urodynamic examination before and after trauma 14 days with catheters inducted into rat bladder per urethra Cystometry、leak point pressure and static urethral pressure profile were recorded.
Results: Ten rats underwent research successfully, The average quantity of maximum bladder volume (MBV)、bladder voiding pressure (VP)、abdomen pressure leak point pressure (ALPP)、maximum urethra pressure (MUP) and maximum urethra closure pressure (MUCP) before injury were ( 0.88±0.46 ) ml、( 26.50±6.42 ) cmH_2O (1cmH_2O=0.098KPa)、(25.65±5.47)cmH_2O、(14.80±2.78)cmH_2O and(13.10±2.60)cmH_2O, respectively; while these parameters were(1.47±0.69)ml、(17.10±6.74)cmH_2O、(15.55±5.24)cmH_2O、(12.10±3.07)cmH_2O and(10.60±3.20)cmH_2O after damage. Compared with those at baseline, the average quantity of MBV increased by (0.60±0.40)ml(P<0.01); bladder VP and ALPP were significantly decreased by (9.40±8.74)cmH_2O and (9.50±4.92)cmH_2O, respectively(P <0.05). MUP and MUCP were decreased by(2.70±3.37)cmH_2O and (2.50±2.59)cmH_2O,respectively(P <0.05). Conclusion: The establishment of animal model for SUI by simulating a birth trauma and the new method of urodynamic examination to evaluate urethral sphincter muscle will make contribution to experimental research of SUI.
Objective: To investigate the expression of insulin - like growth factor-I、II( IGF-I、II) during regeneration following urethral sphincter muscle injury in female Sprague- Dawley (SD) rats. To evaluate the effect of periurethral injection of IGF-I on the expression of organic IGF-I、IGF-II mRNA and the change of morphological Characteristics of urethral sphincter muscle during repair and regeneration period.
Methods: Female virgin SD rats were performed urethral sphincter muscle injury by simulating a birth trauma.①Then the rats (n=25) were killed in different time (2、4、6、8、14day) and specimens were processed for reverse transcription (RT) and polymerase chain reaction (PCR). A control group (n=5) underwent the same procedurewithout intravaginal balloon inflation.②Then the rats (n=50) was divided randomly into 2 subgroups: 25 rats underwent periurethral injection of 1.0μg human IGF-I to the middle urethral muscle and the other 25 rats as control group underwent saline injection. Five rats in each group were killed at 2、4、6、8、14 day respectively and the whole urethra specimens were processed forRT-PCR. A control group (n=5) underwent the same procedur without intravaginal balloon inflation and injection.③Then the rats (n=20) was divided randomly into 2 subgroups: 20 rats underwent periurethral injection of 1.0μg human IGF-I to the middle urethral muscle and the other 20 rats as control group underwent saline injection. Four rats in each group were killed at 2、4、6、8、14 day respectively, the middle one third of the urethra specimens were harvested for step serial histological paraffin sections and used for Masson’s trichrome staining. Pathological changes and the area percentage of the striated muscle, the smooth muscle and the dense connective tissue were assessed with computer- assisted image analysis system. A control group (n=4) underwent the same procedur without intravaginal balloon inflation and injection.
Results:①IGF - I mRNA increased at day 4、6、8、14 after urethral sphincter muscle injury, especially at day 8 (P<0.05);IGF-II mRNA increased at day 4、6、8 after urethral sphincter muscle injury, especially at day 6、8(P<0.05).②After IGF-I injection, the expression of IGF-I mRNA increased significantly at all time points, with a peak on day 8.Comparied with saline control group, the difference was significant on day 4、14(P<0.01), and day 8(P<0.05). No expression on day 2 was noted in saline control group.The expression of IGF-II mRNA in experiment group increased on day 4、6、8、14.Comparied with saline control group,the significant difference noted on day 4(P<0.01).No expression on day 14 was noted in saline control group.③After simulated birth injury, the microstructure of mid-urethra changed significantly, with extensive disruption of the striated muscle layer, significant decline in urethral wall musculature (both smooth and striated), irregular shape and collagen fibers hyperplasia obviously. Comparied with saline control group, the area percentage of the striated muscle and the smooth muscle increased significantly on day 8 and day 14(P<0.05) after IGF-I injection.
Conclusion: IGF-I、II mRNA was expressed during regeneration following urethral sphincter muscle injury and related to the healing process. The expression of organic IGF-I and IGF-II mRNA was improved by periurethral injection of IGF-I during regeneration period following urethral sphincter muscle injury in female rat. The repair of impaired urethral spincter muscle was improved by periurethral injection of IGF-I during regeneration period following urethral sphincter muscle injury in female rat. Our findings suggest that IGF-I facilitates the regeneration of the urethral muscles and may play a role in treatment of stress urinary incontinence induced by urethral sphincter muscle dysfunction.
方法:12只成年雌性未育大鼠在模拟产伤致尿道括约肌损伤前及损伤后第14天经尿道置管行充盈期膀胱测压、漏尿点压力和静态尿道压力测定。
结果10只顺利完成实验,损伤前平均最大膀胱容量、排尿压、腹压漏尿点压、最大尿道压、最大尿道闭合压分别为(0.88±0.46)ml、(26.50±6.42)cmH_2O (1cmH_2O=0.098KPa)、(25.65±5.47)cmH_2O、(14.80±2.78)cmH_2O和(13.10±2.60)cmH_2O,损伤后分别为(1.47±0.69)ml、(17.10±6.74)cmH_2O、(15.55±5.24)cmH_2O、(12.10±3.07)cmH_2O和(10.60±3.20)cmH_2O,大鼠最大膀胱容量增加(0.60±0.40)ml(P<0.01),排尿压、腹压漏尿点压分别下降(9.40±8.74)cmH_2O、(9.50±4.92)cmH_2O(P<0.05),最大尿道压和最大尿道闭合压亦下降(2.70±3.37)cmH_2O、(2.50±2.59)cmH_2O(P<0.05)。
结论模拟产伤可有效建立雌性大鼠压力性尿失禁模型,经尿道置管的尿动力检测方法能客观评价大鼠尿道括约肌功能。
目的:观察雌性Sprague - Dawley(SD)大鼠尿道括约肌损伤后组织修复过程中局部内源性胰岛素样生长因子-I和II的mRNA表达情况。研究胰岛素样生长因子-I局部注射在其损伤后修复过程中对内源性IGF-I和II mRNA表达的影响及对尿道显微形态结构改变的影响。
方法:制作模拟产伤的SD大鼠尿道括约肌损伤模型后,①分别于损伤后2、4、6、8、14天处死大鼠取出尿道,行逆转录-聚合酶链反应(RT- PCR),琼脂糖凝胶电泳,通过辉度扫描与内参照对比,了解IGF– I和II的mRNA表达变化;②于尿道中段局部注射IGF-I,RT-PCR半定量分析损伤后2、4、6、8、14d时IGF-I和II mRNA表达;③于尿道中段局部注射IGF-I,中段尿道Masson’s trichrome染色,观察其显微结构动态改变,并运用计算机图象分析系统定量分析尿道横纹肌、平滑肌及结缔组织组成。尿道括约肌无损伤大鼠为正常对照组。
结果:①IGF - I mRNA在损伤后4、6、8、14天表达,尤以第8天升高明显(P<0.05);IGF-II mRNA在伤后4、6、8天表达,以第6、8天升高明显(P<0.05);未损伤组无明显IGF表达。②试验组内源性IGF-I mRNA在伤后2、4、6、8、14d均有表达,尤以第8d增加明显,与生理盐水对照组比较于第4、14d表达有极显著增加(P<0.01),第8d有显著增加(P<0.05);试验组IGF-II mRNA在伤后4、6、8、14d表达,以第6、8d增加明显,与生理盐水对照组比较第4d有极显著增加(P<0.01)。③损伤后尿道显微结构发生明显改变,尿道横纹肌、平滑肌出现断裂,萎缩,排列结构紊乱,胶原结缔组织增生明显。试验组与生理盐水对照组比较于8d、14d肌组织所占尿道横截面积比例明显增高(P<0.05),肌纤维增生丰富,形态结构更为完整。
结论:SD大鼠尿道括约肌损伤修复过程中有内源性IGF-I和II的mRNA表达变化,提示IGF参与尿道括约肌损伤后肌细胞再生修复过程;局部注射IGF-I能促进尿道括约肌损伤局部内源性IGF-I和II mRNA的表达,并能促进尿道括约肌肌细胞损伤后的修复,使尿道括约肌形态结构更为完整,提示IGF-I对尿道括约肌损伤及其所致压力性尿失禁具有潜在的治疗作用。
Objective: To evaluate the effect of a new method of urodynamics in the research of female rat stress urinary incontinence model and present the urodynamic characteristic of urethral sphincter dysfunction of rat.
Method: Twelve female virgin SD rats were performed urethral sphincter muscle injury by simulating a birth trauma. They all underwent urodynamic examination before and after trauma 14 days with catheters inducted into rat bladder per urethra Cystometry、leak point pressure and static urethral pressure profile were recorded.
Results: Ten rats underwent research successfully, The average quantity of maximum bladder volume (MBV)、bladder voiding pressure (VP)、abdomen pressure leak point pressure (ALPP)、maximum urethra pressure (MUP) and maximum urethra closure pressure (MUCP) before injury were ( 0.88±0.46 ) ml、( 26.50±6.42 ) cmH_2O (1cmH_2O=0.098KPa)、(25.65±5.47)cmH_2O、(14.80±2.78)cmH_2O and(13.10±2.60)cmH_2O, respectively; while these parameters were(1.47±0.69)ml、(17.10±6.74)cmH_2O、(15.55±5.24)cmH_2O、(12.10±3.07)cmH_2O and(10.60±3.20)cmH_2O after damage. Compared with those at baseline, the average quantity of MBV increased by (0.60±0.40)ml(P<0.01); bladder VP and ALPP were significantly decreased by (9.40±8.74)cmH_2O and (9.50±4.92)cmH_2O, respectively(P <0.05). MUP and MUCP were decreased by(2.70±3.37)cmH_2O and (2.50±2.59)cmH_2O,respectively(P <0.05). Conclusion: The establishment of animal model for SUI by simulating a birth trauma and the new method of urodynamic examination to evaluate urethral sphincter muscle will make contribution to experimental research of SUI.
Objective: To investigate the expression of insulin - like growth factor-I、II( IGF-I、II) during regeneration following urethral sphincter muscle injury in female Sprague- Dawley (SD) rats. To evaluate the effect of periurethral injection of IGF-I on the expression of organic IGF-I、IGF-II mRNA and the change of morphological Characteristics of urethral sphincter muscle during repair and regeneration period.
Methods: Female virgin SD rats were performed urethral sphincter muscle injury by simulating a birth trauma.①Then the rats (n=25) were killed in different time (2、4、6、8、14day) and specimens were processed for reverse transcription (RT) and polymerase chain reaction (PCR). A control group (n=5) underwent the same procedurewithout intravaginal balloon inflation.②Then the rats (n=50) was divided randomly into 2 subgroups: 25 rats underwent periurethral injection of 1.0μg human IGF-I to the middle urethral muscle and the other 25 rats as control group underwent saline injection. Five rats in each group were killed at 2、4、6、8、14 day respectively and the whole urethra specimens were processed forRT-PCR. A control group (n=5) underwent the same procedur without intravaginal balloon inflation and injection.③Then the rats (n=20) was divided randomly into 2 subgroups: 20 rats underwent periurethral injection of 1.0μg human IGF-I to the middle urethral muscle and the other 20 rats as control group underwent saline injection. Four rats in each group were killed at 2、4、6、8、14 day respectively, the middle one third of the urethra specimens were harvested for step serial histological paraffin sections and used for Masson’s trichrome staining. Pathological changes and the area percentage of the striated muscle, the smooth muscle and the dense connective tissue were assessed with computer- assisted image analysis system. A control group (n=4) underwent the same procedur without intravaginal balloon inflation and injection.
Results:①IGF - I mRNA increased at day 4、6、8、14 after urethral sphincter muscle injury, especially at day 8 (P<0.05);IGF-II mRNA increased at day 4、6、8 after urethral sphincter muscle injury, especially at day 6、8(P<0.05).②After IGF-I injection, the expression of IGF-I mRNA increased significantly at all time points, with a peak on day 8.Comparied with saline control group, the difference was significant on day 4、14(P<0.01), and day 8(P<0.05). No expression on day 2 was noted in saline control group.The expression of IGF-II mRNA in experiment group increased on day 4、6、8、14.Comparied with saline control group,the significant difference noted on day 4(P<0.01).No expression on day 14 was noted in saline control group.③After simulated birth injury, the microstructure of mid-urethra changed significantly, with extensive disruption of the striated muscle layer, significant decline in urethral wall musculature (both smooth and striated), irregular shape and collagen fibers hyperplasia obviously. Comparied with saline control group, the area percentage of the striated muscle and the smooth muscle increased significantly on day 8 and day 14(P<0.05) after IGF-I injection.
Conclusion: IGF-I、II mRNA was expressed during regeneration following urethral sphincter muscle injury and related to the healing process. The expression of organic IGF-I and IGF-II mRNA was improved by periurethral injection of IGF-I during regeneration period following urethral sphincter muscle injury in female rat. The repair of impaired urethral spincter muscle was improved by periurethral injection of IGF-I during regeneration period following urethral sphincter muscle injury in female rat. Our findings suggest that IGF-I facilitates the regeneration of the urethral muscles and may play a role in treatment of stress urinary incontinence induced by urethral sphincter muscle dysfunction.
引文
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