ARFGAP 3在小鼠盆底肌损伤模型中的表达  

Expression of ARFGAP 3 in mouse model of pelvic floor muscle injury

作　　者：陈茂 洪莉[1] 胡鸣[1] 汤剑明[1] 王琳琳[1] 李素廷 陈乾 李秉枢[1] 洪莎莎[1] CHEN Mao;HONG Li;HU Ming;TANG Jian-ming;WANG Lin-lin;LI Su-ting;CHEN Qian;LI Bing-shu;HONG Sha-sha(Department of Obstetrics and Gynecology,People's Hospital of Wuhan University,Wuhan Hubei 430060,P. R. China)

机构地区：[1]武汉大学人民医院妇产科

出　　处：《中国计划生育和妇产科》2019年第10期84-87,96,I0002,共6页Chinese Journal of Family Planning & Gynecotokology

基　　金：国家自然科学基金资助项目(项目编号:81771562);中央高校基本科研业务费专项资金重大培育类(项目编号:2041018kf0066)

摘　　要：目的探讨ADP核糖基化因子GTP结合蛋白3(ADP ribosylation factor GTPase binding protein 3,ARFGAP 3)在小鼠盆底肌损伤模型中的表达变化以及在压力性尿失禁(stress urinary incontinence, SUI)靶向治疗中的初步分析。方法选择雌性成年处鼠C 57 BL/6小鼠40只,随机分为5组,每组8只。根据造模后取材时间点的不同分为1 d、3 d、7 d、14 d组和对照组。造模采用经典的阴道扩张法模拟产伤构建盆底肌损伤小鼠模型,分别于处理后第1、3、7、14 d处死小鼠,取盆底肌组织进行分析,对照组常规饲养1周后取盆底肌作相同处理。HE染色观察盆底肌病理变化,确定损伤的存在及持续时间;免疫组织化学法和Western-blot法检测ARFGAP 3蛋白水平表达变化;q-PCR检测ARFGAP 3的mRNA表达变化。结果①1 d、3 d组盆底肌损伤变化明显,肌膜间可见大量炎性细胞浸润,部分肌纤维小灶性坏死,肌间隙增宽,肌束结构不清晰;7 d组盆底肌组织损伤趋于修复,少量肌纤维萎缩,炎性浸润减少,结缔组织增生;14 d组小鼠盆底肌纤维、肌束结构与对照组相比基本恢复正常,肌间隙略宽。②免疫组化染色、Western-blot检测以及q-PCR结果相一致,各造模组较对照组小鼠盆底肌组织中ARFGAP 3表达上调(P<0.05),3 d组上调较各造模组最明显。结论①模拟产伤法构建的盆底肌损伤模型中盆底肌损伤明显,成功构建盆底肌损伤小鼠模型,有助于进一步探究SUI发病机制;②小鼠盆底肌损伤后ARFGAP 3表达上调,且于第3 d增高最明显,随后表达逐渐降低,这表明ARFGAP 3可能参与了盆底肌损伤修复过程。Objective To investigate the expression of ADP ribonylation factor GTP ase binding protein 3(ARFGAP 3) in mouse pelvic floor muscle injury model and its preliminary analysis in targeted therapy of stress urinary incontinence(SUI). Methods Forty virgin female adult mice of C57 BL/6 strain were randomized into 5 groups, 8 mice per group: 1 d, 3 d, 7 d and 14 d group and control group according to different sampling time points after modeling.The mice models of pelvic floor muscle injury were constructed by using the classic vaginal dilation method to simulate the birth injury. The mice were sacrificed to obtain pelvic floor muscle tissue for analysis on the first, third, and seventh, 14 th days after treatment, and the control group received pelvic floor muscle for the same treatment after 1 week of routine feeding. Changes of pelvic floor myopathy were observed by HE staining, and to determine the presence and duration of lesions.The expression of ARFGAP 3 protein was detected by immunohistochemistry and western-blot.The mRNA expression of ARFGAP 3 was detected by q-PCR. Results ① There were significant changes in pelvic floor muscle injury of 1 d and 3 d group, a large number of inflammatory cell infiltration could be seen in the muscle membrane, some muscle fiber small focal necrosis, muscle gap widened, and muscle bundle structure was not clear. In 7 d group, the pelvic floor muscle tissue injury tended to repair, with a few muscle fiber atrophy, reduced inflammatory infiltration, and connective tissue hyperplasia. Compared with the control group, the structure of pelvic floor muscle fiber and muscle bundle in 14 d group basically returned to normal, and the muscle gap was slightly wider.② The results of immunohistochemical staining, western-blot and q-PCR were consistent. Compared with the control group, the expression of ARFGAP 3 was up-regulated in the pelvic floor muscle tissues of mice in each model group(P<0.05), and the up-regulated expression was most obvious in the 3 d group.Conclusion ① The pelv

关 键 词：ARFGAP 3 盆底肌损伤 模拟产伤 损伤修复 

分 类 号：R711.4[医药卫生—妇产科学]