小鼠单侧睾丸动脉结扎构建自身免疫性睾丸炎模型的探讨

发布时间：2018-07-01 16:13

本文选题：动脉结扎 + 自身免疫性睾丸炎　；参考：《新疆医科大学》2009年硕士论文

【摘要】： 目的:本实验通过结扎小鼠一侧睾丸动脉并于不同时间后再通以诱导自身免疫性睾丸炎,观察睾丸生精上皮、血液中抗精子抗体(AsAb)IgG、IgM以及迟发型超敏反应的变化。实验探讨了一侧睾丸动脉结扎再通后,小鼠发生自身免疫性睾丸炎的机制,并试图将此作为一种新型的构建小鼠自身免疫性睾丸炎模型的方法。方法:选用健康成年雄性BALB/c小鼠,随机分为4组:对照组(A)(麻醉后仅进行阴囊切开后缝合),实验1组(S1)(单侧睾丸动脉结扎,2小时后再开通血管),实验2组(S2)(单侧睾丸动脉结扎,4小时后开通血管),实验3组(S3)(单侧睾丸动脉结扎,6小时后再开通血管)。分别于术后12h,2周,4周应用光镜观察石蜡包埋HE染色后的小鼠睾丸组织结构,应用透射电镜技术观察术后12小时小鼠睾丸组织的超微结构。采用ELISA法检测2周、4周时各组小鼠血液中的抗精子抗体(AsAb)IgG、IgM浓度水平;采用游标卡尺测量单侧动脉结扎再通后对睾丸自身抗原的足垫反应。结果:1.HE染色观察:①术后12h的,A组小鼠睾丸生精小管内无生精细胞脱落,各级生精细胞排列整齐、层次清楚,结构正常,显示活跃的精子发生。间质中可见间质细胞正常,核大而圆,偏位。S1组小鼠术侧睾丸生精小管较为完整,小管内可见轻微的生精细胞脱落,偶见生精小管内空泡变。S3组小鼠术侧睾丸生精小管之间间隙增宽,生精小管内生精细胞层数减少。间质中血管充血明显,管腔中充满了红细胞,血管壁增厚呈玻璃样变,同时血管内近壁侧可以看见较多的炎性细胞(主要是白细胞)的浸润。S4组小鼠术侧睾丸间质中可以看到较多的炎性细胞浸润。睾丸生精小管内生精细胞层数减少,看不到精子,细胞肿胀明显,胞质透亮。可以看到多核巨细胞散落分布在睾丸生精小管管腔中。各组非术侧睾丸生精上皮规则,生精细胞排列有序,可见大量精子。间质中可见间质细胞,核大而圆,偏位。②术后2周的光镜结果:A组小鼠睾丸组织结构正常,与术后12h无显著差异。S1组睾丸生精小管内生精细胞层数减少,可见空泡状结构。S2、S3组均可见不同程度的生精细胞脱落、排列紊乱,生精上皮变薄、空泡样变。间质中血管增多、血管壁变厚,在血管周围和生精小管界膜附近,可见炎性细胞浸润。③术后4周时各组光镜结果与2周时相比较,无明显结构上的差异。2.透射电镜观察:A组小鼠睾丸生精上皮结构完整,细胞无肿胀变性;其余各手术组均可见不同程度的睾丸生精细胞肿胀变性坏死。3.血清抗精子抗体IgG与IgM检测结果:各实验组与正常对照组比较,血清中抗精子抗体IgG及IgM的浓度明显升高,具有统计学意义(P0.05);4.足趾肿胀实验:各手术组均出现迟发型过敏反应。结论:单侧睾丸动脉结扎/再通后:1.术侧睾丸组织首先表现为一种缺血性的炎症表现,有大量的炎性细胞的浸润(主要是白细胞);2.术后虽然恢复了血液供应,但仍可见小鼠睾丸生精上皮脱落明显,生精细胞层数减少,可见炎细胞浸润(主要为单个核细胞)。3.血清中有抗精子抗体IgG、IgM出现。4.可以刺激机体形成迟发型超敏反应。5.此种手术方法可以成功建立自身免疫性睾丸炎模型。
[Abstract]:Objective: to induce autoimmune orchitis by ligating one of the testicular arteries in mice at a different time and observing the changes in the spermatogenic epithelium of the testis and the changes of anti sperm antibody (AsAb) IgG, IgM and delayed hypersensitivity in the blood. Mechanism, and try to use this as a new model of mouse autoimmune orchitis. Methods: healthy adult male BALB/c mice were randomly divided into 4 groups: control group (A) (after anaesthesia only after scrotal incision suture), experimental 1 (S1) (unilateral testicular artery ligation, 2 hours after the reopening of blood vessels), experimental 2 (S2) (single The lateral testis artery was ligated and the blood vessels were opened 4 hours later. The experimental 3 groups (S3) (unilateral testicular artery ligation and reopening of blood vessels after 6 hours) were used to observe the histological structure of the testis after paraffin embedded HE staining in 12h, 2 weeks and 4 weeks after the operation, and the ultrastructure of the testis in 12 hours after the operation was observed by transmission electron microscopy. ELISA was used to observe the ultrastructure of the testis in the mice. The method was used to detect the level of antisperm antibody (AsAb) IgG and IgM in the blood of each group for 2 weeks and 4 weeks. The foot pad reaction of the autoantigen of the testis was measured by the vernier caliper and after the unilateral artery was ligated and repassed. Results: 1.HE staining was used to observe: (1) after 12h, the no spermatogenic cells in the testicular seminiferous tubules of the A mice were lost, and the spermatogenic cells at all levels were arranged neatly. The interstitial cells were normal, the interstitial cells were normal, the nucleus was large and round, and the testicular seminiferous tubules on the side of the.S1 group were more complete, and the minor spermatogenic cells in the tubule were seen in the tubule. The interstitial cells in the vacuoled.S3 group of the spermatogenic tubule group were widened in the space between the seminiferous tubules and the seminiferous tubules. The number of spermatogenic cells decreased. The vascular congestion was obvious in the interstitium, red blood cells were filled in the lumen, the thickening of the wall of the blood vessel was glass like change. At the same time, more inflammatory cells (mainly leucocytes) in the proximal wall of the blood vessels were seen in the.S4 group. The number of cell layers was reduced, the sperm was not visible, the cells were swollen and the cytoplasm was bright. It could be seen that the multinuclear giant cells scattered in the seminiferous tubules. The rules of the spermatogenic epithelium in the non lateral testis were regular, the spermatogenic cells were arranged in order, and a large number of spermatozoa were visible. The interstitial cells were visible in the interstitium, and the nucleus was large and round and partial. (2) the results of light microscopy 2 weeks after the operation: A The histological structure of the testis was normal in the group of mice. There was no significant difference between the 12h and the postoperatively. The number of spermatogenic cells in the testicular seminiferous tubules in the.S1 group decreased, and the vacuolated structure was.S2. The S3 group showed the exfoliation of spermatogenic cells in different degrees, the disordered arrangement, the thinning of the spermatogenic epithelium and the vacuolation. The blood Guan Zengduo, the blood vessel wall thickened, and the peripheral blood vessels and the seminiferous tubules In the 4 weeks after the operation, the results of the light microscopy were compared with the 2 weeks, and there was no obvious difference in structure.2. transmission electron microscopy: the spermatogenic epithelium of the mice in group A was complete and the cells were not swollen and denatured, and the other surgical groups showed a different degree of testicular spermatocyte swelling and necrosis of.3. serum ANTISPERMATOGENESIS. The results of the detection of subantibody IgG and IgM: compared with the normal control group, the concentration of antisperm antibody IgG and IgM in the serum was significantly increased, with statistical significance (P0.05); 4. toe swelling test: all the operation groups had delayed anaphylaxis. Conclusion: the unilateral testicular vein ligation / repassage: the 1. operative side of the testis is the first manifestation of one kind. There was a large number of inflammatory cells infiltrating (mainly white blood cells). After 2., the blood supply was restored, but the spermatogenic epithelium of mouse testis was obvious, the number of spermatogenic cells decreased, and the infiltration of inflammatory cells (mainly mononuclear cells) had anti sperm antibody IgG in.3. serum, and.4. could be stimulated by IgM. .5. is a delayed type hypersensitivity reaction. This method can successfully establish the autoimmune orchitis model.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R-332;R697.22

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