去甲肾上腺素诱导HSP70的表达对隐睾生殖细胞凋亡的影响

发布时间：2018-05-08 15:03

  本文选题：去甲肾上腺素 + 隐睾　； 参考：《遵义医学院》2013年硕士论文

【摘要】：目的：通过比较去甲肾上腺素诱导HSP70的表达对隐睾症大鼠生殖细胞凋亡的影响,探讨去甲肾上腺素对隐睾症的治疗作用与机制,为隐睾症的临床治疗提供理论实验依据。 方法：22日龄雄性SPF(specific pathogen-free animal)级SD大鼠96只,体重40-55g,经随机数字表处理后随机分为3组：空白组A组(n=32)空白对照组,不采取任何干预措施；剩余两组建立单侧隐睾模型后分别施予不同干预措施,隐睾模型组B组(n=32)隐睾模型生理盐水处理组(按5ml/kg/d剂量腹腔注射给药)；隐睾模型药物处理(用生理盐水配成O.1mg/ml,按0.5mg/kg/d剂量腹腔注射给药)组C组(n=32)。并于建立模型后的当天,第5,10,15d按随机的方法取出各组SD大鼠8只心脏穿刺采血3ml,作血清抗体分析用,然后断颈处死SD大鼠迅速取出隐睾侧睾丸称重后,按常规方法石蜡包埋固定,标记后留做HE染色观查并检测曲细精管生育力指数(tubular fertility index, TFI)、平均曲细精管直径(mean tubular diametar, MTD),免疫组化,凋亡检测用。 结果： (1)各组大鼠隐睾侧睾丸重量的变化：睾丸手术后5d时,B组睾丸重量就明显下降(P0.05),随后持续下降,到15d时睾丸重量仅为A组的35%。C组变化较小,10d开始下降,与A组、B组的差异均有统计学意义(P0.05)。 (2)各组大鼠血清抗精子抗体(AsAb)IgG、IgA、IgM定量测定：睾丸手术后5d时,B组睾丸IgG和IgA水平开始升高(P0.05),且随着时间的延长,升高越明显；而B组睾丸IgM水平手术后5d时就明显上升(P0.05),10d后开始下降。C组变化较小与A组、B组的差异均有统计学意义(P0.05),IgG和IgA水平手术后5d时升高不明显,无统计学意义(P0.05),10d后开始上升,与A组、B组比较,有统计学意义(P0.05),与A组、B组比较,有统计学意义(P0.05),但升高水平较B组低,而IgM水平手术后5d时就明显上升(P0.05),后随时间延长逐渐降低。 (3)各组大鼠组织学观察：B、C组MTD与TFI在手术后5d明显降低,与A比较组有统计学意义(P0.05), C组术后10d后MTD与TFI有所升高,与A组和B组比较,有统计学意义(P0.05)。 (4)各组大鼠细胞凋亡水平变化：B组于隐睾模型建立后在5d时就凋亡明显增加(P0.05),且随时间的延长逐渐增加,C组隐睾模型建立后在5d时就凋亡明显增加(P0.05),且随时间的延长逐渐增加,但无B组明显,与A组、B组比较,均有统计学意义(P0.05)； (5)各组大鼠Hsp70表达量变化：在A组睾丸组织中,Hsp70呈中等程度表达于精母细胞和精子细胞的细胞核内。B组Hsp70于隐睾模型建立后在5d时就明显减少(P0.05),10d时减少达到高峰。到隐睾术后15d时,无明显Hsp70表达；C组Hsp70隐睾模型建立后在5d也明显减少(P0.05),10d后Hsp70表达有所增加,与A组、B组比较,均有统计学意义(P0.05)。 结论： (1)去甲肾上腺素可抑制大鼠手术诱导隐睾后生殖细胞细胞凋亡； (2)去甲肾上腺素可上调大鼠手术诱导隐睾后生殖细胞HSP70的表达。
[Abstract]:Objective: to compare the effect of norepinephrine induced HSP70 expression on germ cell apoptosis in rats with cryptorchidism, to explore the therapeutic effect and mechanism of norepinephrine on cryptorchidism, and to provide theoretical and experimental evidence for clinical treatment of cryptorchidism. Methods Ninety-six male SPF(specific pathogen-free animal- grade SD rats at the age of 22 days, weighing 40-55 g, were randomly divided into three groups: blank group A (group A) and control group (n = 32). The other two groups were given different intervention measures after establishing unilateral cryptorchidism model. The rats in the cryptorchidism model group (group B) were treated with normal saline (intraperitoneal injection of 5ml/kg/d) and group C (treated with 0.1 mg / ml of normal saline and intraperitoneal injection of 0.5mg/kg/d). On the day after the establishment of the model, 8 SD rats in each group were randomly collected for 3 ml of blood by heart puncture on the 15th day after the establishment of the model, and then the SD rats were killed by cervical amputation, and the testis of the cryptorchidism side were quickly removed and weighed. The seminiferous tubules were fixed with paraffin wax and then stained with HE staining. The fertility index of seminiferous tubules was detected by tubule fertility index, and the mean diameter of seminiferous tubules was mean tubular diametar, MTDX, immunohistochemistry, apoptosis detection. Results: (1) the changes of testicular weight in the cryptorchidism side of rats in each group: the testis weight of group B significantly decreased at 5 days after testicular operation, and then continued to decrease, and at 15 days the weight of testis in group A was only 35.C, the change of testicular weight was less than that in group A and the change began to decrease at 10 days after testicular surgery. The difference between group A and group B was statistically significant (P 0.05). (2) quantitative determination of serum anti-sperm antibody (Asab) IgG IgG and IgA in rats in each group: the levels of IgG and IgA in testis of group B began to increase at 5 days after testicular operation, and the increase was more obvious with the prolongation of time. However, the level of IgM in testis of group B increased significantly 5 days after operation and began to decrease 10 days after operation. The difference between group C and group A was statistically significant. The levels of IgM and IgA in group B were not significantly increased at the 5th day after operation, but they began to rise after 10 days without statistical significance. Compared with group A and group B, there was significant difference between group A and group B (P 0.05), but the level of elevation was lower than that of group B, but the level of IgM increased significantly at 5 days after operation, and then decreased gradually with the prolongation of time. (3) histological observation showed that MTD and TFI in group C were significantly lower than those in group A on the 5th day after operation, and there were significant differences between group A and group A (P 0.05), MTD and TFI in group C were increased 10 days after operation, compared with group A and group B, there was significant difference between group A and group B (P 0.05). (4) the apoptosis level of rats in group B increased significantly at the 5th day after the establishment of the cryptorchidism model, and gradually increased with the extension of time, and the apoptosis of the cryptorchidism model in group C increased significantly at the 5th day after the establishment of the model, and gradually increased with the extension of time. But there was no significant difference between group B and group A (P 0.05). (5) the changes of Hsp70 expression: in group A, the expression of Hsp70 in spermatocytes and spermatocytes was moderate. Group B showed a significant decrease in the expression of Hsp70 in the nucleus of spermatocytes and spermatozoa at 5 days after the establishment of the cryptorchidism model, and the decrease of Hsp70 reached its peak at 10 days after the establishment of the cryptorchidism model. On the 15th day after operation, the expression of Hsp70 in group C was significantly decreased after the establishment of the model of cryptorchidism without Hsp70 expression. The expression of Hsp70 in group C was significantly lower than that in group A (P 0.05) 10 days after operation, and there was significant difference between group A and group B (P 0.05). Conclusion: 1) norepinephrine could inhibit the germ cell apoptosis induced by operation in rat cryptorchidism. (2) norepinephrine could up-regulate the expression of HSP70 in germ cells of rats after operation induced by cryptorchidism.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R726.9

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