NOS在压力性尿失禁大鼠盆底组织中的表达及意义  

Expression and significance of NOS in pelvic floor tissue of stress urinary incontinence rats

作　　者：苟鑫 吴静 罗伟 柯健[1] 杨智明[1] 袁志伟[1] 孙浩 杨昭庆 方克伟[1] GOU Xin;WU Jing;LUO Wei;KE Jian;YANG Zhiming;YUAN Zhiwei;SUN Hao;YANG Zhaoqing;FANG Kewei(Department of Urology,the Second Affiliated Hospital of Kunming Medical University,Kunming,Yunnan 650101,China;Department of Biochemistry and Molecular Biology,Basic Medical College,Kunming Medical University,Kunming,Yunnan 650500,China;Department of Genetics,Institute of Medical Biology,Chinese Academy of Medical Sciences,Kunming,Yunnan 650031,China)

机构地区：[1]昆明医科大学第二附属医院泌尿外科,650101 [2]昆明医科大学基础医学院生物化学和分子生物学系,650500 [3]中国医学科学院医学生物学研究所遗传室,昆明650031

出　　处：《重庆医学》2020年第18期2984-2990,共7页Chongqing medicine

基　　金：国家自然科学基金项目(80860127);云南省科技计划项目(2015 FA007);云南省高校女性盆底疾病诊疗工程研究中心;云南省科学和技术带头人后备人才项目(2017HB038);云南省卫生和计划生育委员会医学学科带头人项目(D-201615)。

摘　　要：目的分析压力性尿失禁(SUI)大鼠阴道、尿道及肛提肌组织中一氧化氮合酶(NOS)的表达情况,探讨NOS在女性SUI发生发展过程中的表达意义。方法以60只健康雌性未育SD大鼠和15只健康雄性SD大鼠为实验对象,在笼中进行自然妊娠,实验分3组,取正常妊娠分娩后的雌性大鼠作为对照组(A组),另有雌性大鼠分娩后扩张阴道组(B组)、阴道扩张+卵巢切除组(C组),每组6只。喷嚏实验及尿动力学检查验证模型,q-PCR、Western blot检测NOS mRNA及蛋白的表达水平,免疫组织化学检测NOS在阴道、尿道及肛提肌组织中的表达。结果实验所得数据经Shapiro-Wilk统计分析,验证均符合正态分布,B组、C组漏尿点压(ALPP)低于A组,其中C组较A组明显下降(P<0.05)。B组、C组与A组比较,衍生型NOS(iNOS)、内皮型NOS(eNOS)、神经元型NOS(nNOS)mRNA及β蛋白表达水平均显著上调,差异有统计学意义(P<0.01)。免疫组织化学结果显示,A组、B组、C组iNOS的染色强度积分在各组织中分别为:尿道上皮(2.66±0.51)、(5.33±0.51)、(9.66±0.51)分,阴道上皮(0.66±0.51)、(5.00±0.63)、(9.33±0.81)分,肛提肌(0.66±0.51)、(5.66±0.51)、(9.33±0.81)分,各组织中组间差异均有统计意义(P<0.05)。eNOS在3组阴道上皮、尿道上皮组织中无表达,在肛提肌组织染色强度积分表达依次为(0.83±0.75)、(5.33±0.51)、(10.00±0.63)分,组间差异有统计意义(均P<0.05)。nNOS在3组尿道上皮组织无表达,在阴道上皮组织中染色强度积分依次为(0.66±0.51)、(5.83±0.98)、(9.66±1.03)分,在肛提肌组织的染色强度积分依次为(0.50±0.54)、(5.50±0.54)、(9.66±1.03)分,组间两两比较差异有统计学意义(均P<0.05)。结论雌性大鼠在卵巢切除后出现SUI症状,而卵巢切除后雌激素下降可致NOS(iNOS、nNOS、eNOS)表达的升高,NOS与SUI的发病可能有关。Objective To analyze the expression of nitric oxide synthase(NOS)in the vagina,urethra and levator ani tissue of female stress urinary incontinence(FSUI)rats,and to explore the expression significance of NOS in the occurrence and development of FSUI.Methods In this study,60 healthy female SD rats and 15 male SD rats were selected as experimental subjects for natural pregnancy in cages,female rats were divided into 3 groups,normal pregnant delivery rats(group A),rats took vaginal dilatation after delivery(group B),rats took vaginal dilatation+ovariectomized(group C),6 rats in each group.The sneeze test and urodynamics test were used to verify the success of SUI modelling.The expression of NOS mRNA and protein in SUIrats′vagina,urethra and levator ani tissue were detected by q-PCR and Western blot,and the expression level of NOS was detected by immunohistochemistry method.Results The experimental data were verified to be normal distribution by Shapiro-Wilk statistical test,ALPP in group B and group C was lower than that in group A,and group C was significantly lower than that in group B(P<0.05).The expressions levels of iNOS,eNOS,and nNOS in group B and group C were significantly increased when compared with group A,and the difference were statistically significant(P<0.01).Immunohistochemical results showed that the staining intensity scores of iNOS in groups A,B,and C were:urethral(2.66±0.51,5.33±0.51,9.66±0.51),vaginal(0.66±0.51,5.00±0.63,9.33±0.81),levator ani tissues(0.66±0.51,5.66±0.51,9.33±0.81),the differences among the three groups were statistically significant(P<0.05).eNOS showed no statistical differences in the vagina and urethra among three groups(P=0.342,P=0.492),and in the levator ani tissues were(0.83±0.75,5.33±0.51,10.00±0.63),there were statistical differences in three groups(P<0.05).There were no expression of nNOS in urethral of the three groups(P=0.629),but it had statistical differences in the vaginal(0.66±0.51,5.83±0.98,9.66±1.03),and levator ani tissues(0.50±0.54,5.50±0.54

关 键 词：尿失禁 压力性 一氧化氮合酶 SD大鼠 模型 盆底组织 

分 类 号：R711.5[医药卫生—妇产科学]