3种睾丸炎小鼠模型建立方法的比较与评价  

Testicular injection of lipopolysaccharide: An effective method for establishing the mouse model of orchitis

作　　者：曹源 颉鸿笙[1,3] 俞文君 郭祺锟 田杜雪 俞心波 蓝立昊 李冬水 况晓东[1] CAO Yuan;XIE Hong-sheng;YU Wen-jun;GUO Qi-kun;TIAN Du-xue;YU Xin-bo;LAN Li-hao;LI Dong-shui;KUANG Xiao-dong(Faculty of Pathology,Nanchang University School of Medicine,Nanchang,Jiangxi 330006,China;TheFirst School of Clinical Medicine,Nanchang University,Nanchang,Jiangxi 330001,China;The Second Schoolof Clinical Medicine,Nanchang University,Nanchang,Jiangxi 330001,China;Queen Mary School of Medicine,Nanchang University,Nanchang,Jiangxi 330001,China;Department of Reproduction,The First Affiliated Hospital of Nanchang University,Nanchang,Jiangxi 330006,China)

机构地区：[1]南昌大学医学院病理教研室,江西南昌330006 [2]南昌大学第一临床医学院,江西南昌330001 [3]南昌大学第二临床医学院,江西南昌330001 [4]南昌大学玛丽女王学院,江西南昌330001 [5]南昌大学第一附属医院生殖科,江西南昌330006

出　　处：《中华男科学杂志》2019年第8期675-680,共6页National Journal of Andrology

基　　金：国家自然科学基金(81460002);江西省自然科学基金(20142BAB205011);江西省教育厅科学技术研究项目(14097)~~

摘　　要：目的:通过3种方法建立昆明小鼠睾丸炎动物模型,比较各模型建立方法的成功率和稳定性并观察睾丸炎性体活化情况,为睾丸炎相关研究提供可靠的模型。方法:40只成年雄性昆明小鼠随机平均分为4组,假手术组、腹腔注射脂多糖组(腹腔LPS组)、手术单侧睾丸注射冰醋酸组(睾丸冰醋酸组)、手术单侧睾丸注射脂多糖组(睾丸LPS组)。使用计算机辅助精液分析(CASA)检测附睾精子浓度及运动参数,HE染色观察睾丸组织的病理改变,Western印迹检测NLRP3炎性体活化相关蛋白Caspase-1、白介素1β(IL-1β)表达情况。结果:与假手术组附睾内精子浓度[(28.20±1.63)×10^6/ml]相比,腹腔LPS组[(25.74±3.19)×10^6/ml]、睾丸冰醋酸组[(17.16±4.41)×10^6/ml]和睾丸LPS组[(16.92±7.13)×10^6/ml]明显降低(P<0.05或P<0.01),且腹腔LPS组与睾丸LPS组之间差异显著(P<0.01)。与假手术组附睾内前向运动精子百分率[(59.34±1.10)%]相比,腹腔LPS组[(29.57±2.16)%]、睾丸冰醋酸组[(18.10±2.38)%]和睾丸LPS组[(7.34±1.63)%]明显下降(P均<0.01),且腹腔LPS组、睾丸冰醋酸组与睾丸LPS组之间差异显著(P均<0.01)。光镜下,可见各实验组睾丸形态结构均发生不同程度病理改变,其中以睾丸LPS组最为显著,睾丸冰醋酸组次之,腹腔LPS组不及前两者。Western印迹结果显示,与假手术组Caspase-1、IL-1β相比,腹腔LPS组、睾丸冰醋酸组及睾丸LPS组表达均明显上调(P均<0.01),且腹腔LPS组与睾丸LPS组之间差异显著(P<0.05)。结论:睾丸注射LPS是3种改良的睾丸炎模型中最有效的,其次是睾丸注射冰醋酸。此外,睾丸炎症的发生可能与NLRP3炎性体活化有关。Objective:To search for a method of establishing a reliable mouse model of orchitis and investigate the association of orchitis with the activation of the inflammasome.Methods:We equally randomized 40 adult male KM mice into groups A (sham operation),B (intraperitoneal injection of lipopolysaccharide [LPS]),C (unilateral testicular injection of glacial acetic acid [GAA]),and D (unilateral testicular injection of LPS).At 3 weeks after modeling,we measured the sperm concentration and percentage of progressively motile sperm (PMS) in the epididymis by computer-assisted semen analysis,observed the pathological changes in the testis tissue by HE staining,and determined the expressions of the Caspase-1 and interleukin (IL)-1β proteins by Western blot. Results:The sperm concentration in the epididymis was significantly decreased in groups B ([25.74 ± 3.19]× 10^6 /ml),C ([17.16 ± 4.41]× 10^6 /ml) and D ([16.92 ± 7.13]× 10^6 /ml) as compared with that in group A ([28.20 ± 1.63]× 10^6 /ml)(all P < 0.05),even more significantly in B than in C and D (P < 0.01),and so was PMS in groups B ([29.57 ± 2.16]%),C ([18.10 ± 2.38]%) and D ([7.34 ± 1.63]%) in comparison with group A ([59.34 ± 1.10]%)(P < 0.01),even more significantly in B and C than in D (P < 0.01).Light microscopy revealed different degrees of pathological changes in the testis tissue,most significant in group D,followed by C and B.Both the expressions of Caspase-1 and IL-1β were remarkably up-regulated in groups B,C and D compared with those in group A (P < 0.01),even more markedly in D than in B and C (P < 0.05).Conclusion:Unilateral testicular injection of LPS is a more efficient method than either unilateral testicular injection of GAA or intraperitoneal injection of LPS for establishing the mouse model of orchitis.Orchitis may be pathologically associated with the activation of the NLRP3 inflammasome.

关 键 词：睾丸炎 男性不育 动物模型 小鼠 

分 类 号：R697.2[医药卫生—泌尿科学]