异常黏液质型阳痿病证大鼠生精功能改变的机制研究

发布时间：2018-09-02 09:49

【摘要】：目的:检测异常黏液质型阳痿病证大鼠睾丸组织中ABP、StAr等生精功能相关关键指标和凋亡情况,并结合睾丸的形态学观察,研究该病证大鼠伴发少精症的病理生理学机制与对证维药伊木萨克片可能的作用机理。方法:取90只雄性、性功能正常的SD大鼠,随机取10只为正常对照组(N),余80只为造模组,采用芫荽实+菠菜实+湿寒性环境的干预条件建立异常黏液质证候模型,动态观测大鼠生物学表征与勃起功能的改变,造模20w后通过APO勃起实验和性行为学实验筛选出阳痿病证模型后,随机分为病证模型组(A1)及病证药物反证组(A31),将未成阳痿者随机分为证候模型组(B1)和证候药物反证组(B3),反证2w后取大鼠睾丸组织和外周血做一下检测:(1)用HE染色观察睾丸组织形态学结构变化,用电镜观察生精细胞形态学结构变化;(2)用免疫组化法检测大鼠睾丸组织ABP蛋白和StAr蛋白的变化;(3)用TUNEL法检测各组大鼠睾丸间质细胞和生精细胞凋亡情况。结果:(1)HE染色睾丸生精细胞形态学结构观察结果显示,N组结构完好;B1、A1组管壁上生精细胞数量明显少于N组,间质细胞及精子数量减少;分别与B1、A1组相比B3、A3组生精小管形态结构改善,各级生精细胞数量有升高趋势。(2)HE染色结果显示:B1、A1组大鼠睾丸间质细胞形态结构与N组比较呈现不规则,水肿,颜色较深,细胞数量少。B3、A3组睾丸间质细胞数量与B1、A1组相比有回升趋势。(3)电镜结果显示:N组睾丸精曲小管结构完整。B1、A1组大鼠睾丸精曲小管形态结构与N组相比呈现紊乱、各级生精细胞数量明显减少,排列不齐,支持细胞现粗面内质网扩张现象。B3、A3组大鼠睾丸生精细胞分界明显,,但管腔内生精细胞排列数量少、部分支持细胞仍然能见到水肿样改变。(4)TUNEL法检测睾丸生精细胞凋亡结果显示:与N组相比B1、A1组细胞凋亡明显增多(P0.01)。B3、A3组与B1、A1组相比细胞凋亡显著下降(P0.01)。(5)TUNEL法检测睾丸间质细胞凋亡结果显示:与N组相比B1、A1组细胞凋亡明显增多(P0.01)。B3、A3组与B1、A1组相比细胞凋亡显著下降(P0.01)。(6)免疫组化结果显示:与N组相比B1、A1组大鼠StAr蛋白表达明显下调。与B1组相比B3组大鼠睾丸StAr蛋白表达显著增多(P0.01)。A3组大鼠睾丸StAr蛋白表达与A1组无显著差异(P0.05)。(7)免疫组化结果显示,B1、A1组大鼠睾丸ABP蛋白表达下调(P0.01)。B3、A3组大鼠睾丸ABP蛋白表达显著上调(P0.05)。结论(1)维医异常黏液质型阳痿病证大鼠睾丸形态学结构发生改变和生精细胞的凋亡可能是其发生少精症的病理学基础。(2)异常黏液质型阳痿病证大鼠外周血T水平下降和睾丸组织ABP、StAr蛋白的下调可能是大鼠出现生精功能改变的重要环节之一。(3)伊木萨克片可能通过多个药物靶点来改善睾丸形态学结构而影响异常黏液质型阳痿病证大鼠生精功能。
[Abstract]:Objective: to detect the key indexes and apoptosis of spermatogenic function such as ABP,StAr in testis of rats with abnormal mucinous impotence and observe the morphology of testis. To study the pathophysiological mechanism of oligozoospermia and the possible action mechanism of Yimusak tablet. Methods: 90 male SD rats with normal sexual function were randomly selected as control group, and 80 (N), rats as model group. The model of abnormal mucus syndromes was established under the intervention conditions of coriander spinach wet and cold environment. The biological characteristics and erectile function of rats were observed dynamically, and the impotence syndrome model was selected by APO erectile test and sexual behavior experiment 20 weeks later. The patients without impotence were randomly divided into syndrome model group (B1) and syndrome drug negative syndrome group (B3). The testis and peripheral blood of rats were taken for 2 weeks. (1) observe by HE staining. Observing the morphological changes of testis, Morphological changes of spermatogenic cells were observed by electron microscope; (2) changes of ABP and StAr proteins in testis were detected by immunohistochemical method; (3) apoptosis of interstitial cells and spermatogenic cells in testis were detected by TUNEL method. Results: (1) the morphological structure of testicular spermatogenic cells was observed by HE staining. The results showed that the number of spermatogenic cells on the wall of group A 1 was less than that of group N, and the number of interstitial cells and spermatozoa decreased. The morphology and structure of seminiferous tubules were improved and the number of spermatogenic cells increased in group B _ (1) A _ (1) compared with group B _ (1) A _ (1) respectively. (2) the results of HE staining showed that the morphology and structure of interstitial cells of testis in group B _ (1) A _ (1) were irregular, edematous and darker than those in group N. The number of testicular mesenchymal cells in the small number of cells. B _ (3) A _ (3) compared with B _ (1) A _ (1) group, the results of electron microscope showed that the testicular seminiferous tubules in group B _ (1) N had complete structure. The morphological structure of testicular seminiferous tubules in group B _ (1) A _ (1) was disordered compared with that in group N. The number of spermatogenic cells at all levels decreased obviously, and the number of spermatogenic cells in the testis of the Sertoli cells expanded in rough endoplasmic reticulum was obvious, but the number of spermatogenic cells in the lumen was less. Some Sertoli cells can still be seen edematous changes. (4) the results of testicular spermatogenic cell apoptosis detected by TUNEL method: compared with the N group, the apoptosis of B1OA1 group was significantly increased (P0.01). B3A3 group was significantly lower than B1A1 group (P0.01). (5) TUNEL method was used to detect the apoptosis of testicular spermatogenic cells (P0.01). (5). The results of cytoplasmic cell apoptosis showed that the apoptosis of B1A1group was significantly higher than that of group N (P0.01). The expression of StAr protein in B1A1group was significantly decreased compared with that in B1A1group (P0.01). (6). Compared with group B1, the expression of StAr protein in testis of B3 group was significantly increased (P0.01). The expression of StAr protein in testis of group A3 was not significantly different from that of group A1 (P0.05). (7). The results of immunohistochemistry showed that the expression of ABP protein down-regulated (P0.01). B3A3 group significantly up-regulated the expression of ABP protein (P0.05). Conclusion (1) the morphological changes of testis and the apoptosis of spermatogenic cells may be the pathological basis of oligozoospermia in rats with abnormal mucinous impotence syndrome. (2) T level in peripheral blood of rats with abnormal mucinous impotence syndrome. The decrease of ABP,StAr protein and the down-regulation of testicular ABP,StAr protein may be one of the important links of spermatogenic function changes in rats. (3) impotence of abnormal mucinous type may be affected by multiple drug targets to improve the morphological structure of testis. The spermatogenic function of rats with disease syndrome.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R29

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