地佐辛长时程应用对雄性小鼠睾丸生精功能的影响

发布时间：2018-01-31 22:24

本文关键词： 地佐辛吗啡睾酮黄体生成素卵泡刺激素　出处：《南华大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：探讨地佐辛(dezocine)长时程应用对雄性小鼠生育能力的影响;初步探讨地佐辛(dezocine)长时程应用导致雄性小鼠生殖功能损害的可能机制。方法：实验选用6周龄C57雄性小鼠60只，体重18~20g，随机分为5组，每组12只，分别为阴性对照组(对照组)，阳性对照组吗啡组(M组)，地佐辛D1(15mg/kg)、D2(20mg/kg)、D3(25mg/kg)组。小鼠适应性饲养1周后开始腹腔注射给药。实验过程中，每天给药之后即刻及用药6小时后观察小鼠一般情况；于实验第21天停止给药，，眼球取血，全自动生化分析仪检测血T、LH及FSH水平；取睾丸组织常规石蜡组织切片，HE染色观察小鼠睾丸病理形态学改变，免疫组织化学染色观察促凋亡因子Bax及凋亡抑制因子Bcl-2在睾丸组织中的表达变化。结果： 1.对照组小鼠腹腔注射后行为正常，其他各组小鼠腹腔注射后均出现兴奋性行为改变，表现为竖尾、奔跑、爬笼等。注射6小时后再观察，各组小鼠行为学表现均恢复正常。 2.HE染色结果显示，对照组睾丸生精小管结构完整、上皮细胞排列整齐，细胞层次清晰，各级生精细胞形态正常，可见大量成熟精子；M组睾丸组织基底膜不完整、生精小管层次紊乱、生精细胞层减少。D组睾丸组织基底膜破坏；生精细胞层排列紊乱、层数减少、细胞脱落、以精原细胞为主，可见精原细胞空泡变性；各级精母细胞减少、变性；精子畸形。随着地佐辛剂量的增加，生精小管受损更为严重，地佐辛对于小鼠生精小管结构的破坏存在剂量依赖性。 3.睾丸组织Bax的表达变化：M组，D1、D2、D3组均明显高于对照组(p＜0.05),D3组与M组相比，Bax的表达显著增加，差异有统计学意义(p＜0.05)；睾丸组织Bcl-2的表达变化：各用药组明显低于对照组(p＜0.05)；D1、D2、D3组间差异有显著性(p＜0.05)，呈现剂量依赖性的表达降低； Bcl-2在D3组的表达显著低于在M组的表达(p＜0.05),差异有统计学意义。 4.D3组小鼠血清卵泡刺激素(FSH)、睾酮(T)、黄体生成素(LH)的表达均低于对照组，差别有统计学意义(p＜0.05)；D3组FSH、T、LH的表达与M组相比，差异有显著性(p0.05)。结论：地佐辛可以导致小鼠出现兴奋表现，对小鼠睾丸组织结构造成破坏，导致Bax和Bcl-2蛋白表达失衡，影响小鼠生殖功能。因此，在临床应用中应该引起广泛关注。
[Abstract]:Objective: To investigate the effect of long-term application of dizocine on the fertility of male mice. To explore the possible mechanism of reproductive dysfunction in male mice induced by long term application of dizocine. Methods: Sixty 6-week-old C57 male mice were randomly divided into 5 groups with 12 rats in each group (control group, morphine group, M group). The mice were given intraperitoneal injection of dizosin D _ (1) / kg ~ (15 mg / kg) ~ (20 mg 路kg ~ (-1) 路kg ~ (-1)) ~ (25 mg / kg)) after adaptive feeding for one week. The general situation of mice was observed immediately after administration every day and 6 hours after administration. On the 21st day of the experiment, the administration was stopped, blood was taken from the eyeball, and the levels of TLH and FSH in blood were detected by automatic biochemical analyzer. The pathological changes of testis in mice were observed by HE staining. Immunohistochemical staining was used to observe the expression of apoptosis-promoting factor Bax and apoptosis-inhibiting factor Bcl-2 in testis. Results: 1. The behavior of mice in the control group was normal after intraperitoneal injection, and the excitatory behavior of other mice was changed after intraperitoneal injection, showing as vertical tail, running, crawling cage and so on. After 6 hours of injection, the changes of excitatory behavior were observed. The behavior of each group of mice returned to normal. 2. The results of HE staining showed that the structure of testicular seminiferous tubules in the control group was intact, the epithelial cells were arranged neatly, the layers of cells were clear, the morphology of spermatogenic cells was normal, and a large number of mature sperm were observed. In group M, the basement membrane of testis was incomplete, the layer of seminiferous tubules was disordered, and the layer of spermatogenic cells was decreased. The layers of spermatogenic cells were disordered, the number of layers decreased, the cells fell off, mainly spermatogonium cells, the vacuolar degeneration of spermatogonium cells was observed. Reduction and degeneration of spermatocytes at all levels; Sperm malformation. With the increase of dizosin dose, the damage of seminiferous tubules was more serious, and the destruction of seminiferous tubules in mice was in a dose-dependent manner. 3. The expression of Bax in testis was significantly higher in group D 1 and D 2 D 3 than that in control group P < 0 05 and group D 3 compared with that in group M, and the expression of Bax in group D 1 and D 2 D 3 was significantly higher than that in group M (P < 0 05). The difference was statistically significant (P < 0.05). The changes of Bcl-2 expression in testicular tissue: the expression of Bcl-2 in each treatment group was significantly lower than that in the control group (P < 0.05); There was a significant difference between the two groups (P < 0.05), and the expression was decreased in a dose-dependent manner. The expression of Bcl-2 in group D 3 was significantly lower than that in group M (P < 0.05). 4. The expression of FSH, TX and LH in D3 group was lower than that in control group (P < 0.05). There was a significant difference in the expression of FSHT LH between group D 3 and group M (P 0.05). Conclusion: Dizosin can cause excitatory performance in mice, damage the structure of mouse testis, lead to the imbalance of Bax and Bcl-2 protein expression, and affect the reproductive function of mice. It should be paid more attention to in clinical application.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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