恐惧应激对雄性大鼠生殖内分泌及生精细胞凋亡的影响

发布时间：2018-05-13 02:11

本文选题：恐惧 + 应激　；参考：《南京师范大学》2007年硕士论文

【摘要】： 应激(Stress)是指机体在受到各种内外环境因素刺激时所发生的非特异性全身反应。长期处于应激状态，机体的自稳态将改变，从而诱发机体产生疾病。遭受战争环境刺激的军人丧失了应付能力，并伴有精神障碍，对此称之为作战应激反应(combat stress reaction CRS)；发展到后期会导致创伤后应激性障碍(post-traumatic stress disorder，PTSD)。本实验建立战争恐惧应激大鼠模型，检测恐惧应激对生殖的影响。研究表明急性应激组与对照组相比，大鼠体重、睾丸重量、附睾重量、精子数量没有明显变化(P＞0.05)，睾丸HE染色显示睾丸结构无明显变化，免疫组化显示3β-HSD表达降低，Western blotting分析3β-HSD降低了7％；慢性应激组与对照组相比，大鼠体重、睾丸重量、附睾重量、精子数量都有显著差异，P＜0.05。睾丸HE染色显示睾丸内生精小管细小，管腔内生精细胞稀少，免疫组化显示3β-HSD表达降低，Western blotting分析3β-HSD降低了19.6％。因此表明随着应激时间的延长生殖器官功能降低。此外，我们用酶联免疫吸附法检测应激组与对照组大鼠血清FSH、LH和T水平，发现急性应激组大鼠血清FSH(2.7±0.4)ng／ml、LH(3.1±0.3)ng／ml和T(3.9±0.5)ng／ml水平略高于对照组大鼠FSH(2.6±0.2)ng／ml、LH(2.8±0.6)ng／ml和T(2.7±0.2)ng／ml水平，但无显著差异(P＞0.05)；亚急性应激组大鼠血清FSH(1.9±0.5)ng／ml、LH(2.9±0.3)ng／ml和T(1.9±1.1)ng／ml水平都低于其对照组大鼠的FSH(2.3±0.4)ng／ml、LH(3.0±0.5)ng／ml和T(2.9±1.3)ng／ml水平，但都无显著差异(P＞0.05)；慢性应激组大鼠血清FSH(1.6±0.5)ng／ml、LH(2.1±0.4)ng／ml和T(0.6±0.3)ng／ml都低于对照组大鼠血清FSH(2.2±0.3)ng／ml、LH(3.0±0.8)ng／ml和T(3.0±0.4)ng／ml，其中大鼠血清LH水平的变化有显著差异(P＜0.05)；而T水平的变化有极显著差异(P＜0.001)。提示恐惧应激会扰乱大鼠的正常生殖内分泌功能。 Annexin5一种钙依赖的磷脂酶结合蛋白，在睾丸支持细胞和间质细胞中也有表达。我们研究表明急性应激组与对照组相比，免疫组化显示Annexin5都定位在间质细胞、支持细胞和成熟的精子头部，Western blotting分析发现Annexin5的表达降低了10.8％；慢性应激组与对照组相比，免疫组化结果显示Annexin5在精子头部的定位逐渐减少，应激1w和2w组，未见到Annexin5定位于精子头部，到了应激的3w和4w，发现Annexin5重新定位于精子的头部，而在间质细胞和支持细胞上的定位模式没有显著变化。Western blotting分析发现Annexin5的表达降低了17.4％。我们用RT-PCR检测了Annexin5mRNA的表达。应激组与各自的对照组相比，Annxin5 mRNA的表达都低于对照组，其中亚急性组与对照组相比有显著差异(P＜0.05)。随着应激时间的延长，Annexin5 mRNA由降低逐渐向生理水平回升。这些数据表明，应激改变大鼠睾丸中Annexin5的正常表达模式。最后，我们还用RT-PCR检测了大鼠睾丸Caspase-3 mRNA和Bax／Bcl-2 mRNA表达的变化。急性应激组与对照组相比，Caspase-3 mRNA的表达水平无显著变化(P＞0.05)，而亚急性应激组、慢性应激组与其对照组相比都有显著差异(P＜0.05)。与对照组相比，急性应激组Bax／Bcl-2 mRNA比值无显著变化(P＞0.05)，而亚急性应激组有显著差异(P＜0.05)，慢性应激组有极显著差异(P＜0.001)。TUNEL结果表明急性应激组与对照组相比，生殖细胞的凋亡指数无显著差异(P＞0.05)，而亚急性应激组和慢性应激组与对照相比生殖细胞凋亡指数有极显著差异(P＜0.001)。因此，长期恐惧应激可导致大鼠睾丸生精细胞发生凋亡，而这种凋亡有Caspase-3及bax和bcl-2等的参与。恐惧应激作为心理应激的一种形式对啮齿类动物睾丸的结构和功能以及生殖内分泌等都有着很大的破坏和干扰作用，并且促使睾丸生精细胞的凋亡，，提示恐惧应激对人类生殖可能有相似的影响。因此，如何减少恐惧应激损伤，对于维持正常生殖功能有着重要的意义。
[Abstract]:Stress ( Stress ) refers to the non - specific systemic reaction that occurs when the organism is stimulated by various internal and external environmental factors . In the long term , the homeostasis of the organism will change , thus inducing the organism to produce disease . The soldier suffering from the war environment loses its ability to cope and is accompanied by mental disorder , which is called the combat stress reaction CRS ;
Development to a later stage may lead to post - traumatic stress disorder ( s ) .

The effects of fear stress on reproduction were studied in this experiment . The results showed that the weight of the testis , the weight of the testis , the weight of the testis and the number of sperms were not significantly changed ( P > 0.05 ) . The results showed that the testis structure was not changed significantly ( P > 0.05 ) , and the expression of 3尾 - HSD was decreased by Western blotting , and the 3 尾 - HSD was reduced by 7 % by Western blotting .
Compared with the control group , the weight of the rat , the weight of the testis , the weight and the number of sperms were significantly different in the chronic stress group ( P < 0.05 ) .

The levels of serum FSH , LH and T were measured by enzyme linked immunosorbent assay ( ELISA ) , and the levels of FSH ( 2.7 卤 0.4 ) ng / ml , LH ( 3.1 卤 0.3 ) ng / ml and T ( 3.9 卤 0.5 ) ng / ml in acute stress group were slightly higher than those in control group ( 2.6 卤 0.2 ) ng / ml , LH ( 2.8 卤 0.6 ) ng / ml and T ( 2.7 卤 0.2 ) ng / ml , but there was no significant difference ( P > 0.05 ) .
The levels of FSH ( 1.9 卤 0.5 ) ng / ml , LH ( 2.9 卤 0.3 ) ng / ml and T ( 1.9 卤 1.1 ) ng / ml in subacute stress group were lower than those in control rats ( 2.3 卤 0.4 ) ng / ml , LH ( 3.0 卤 0.5 ) ng / ml and T ( 2.9 卤 1.3 ) ng / ml , but there was no significant difference ( P > 0.05 ) .
The serum levels of FSH ( 1.6 卤 0.5 ) ng / ml , LH ( 2.1 卤 0.4 ) ng / ml and T ( 0.6 卤 0.3 ) ng / ml in chronic stress group were lower than those in control group ( 2.2 卤 0.3 ) ng / ml , LH ( 3.0 卤 0.8 ) ng / ml and T ( 3.0 卤 0.4 ) ng / ml , respectively .
There was significant difference in T level ( P < 0 . 001 ) . It was suggested that fear stress could disturb the normal reproductive endocrine function of rats .

Annexin5 , a calcium - dependent phospholipase binding protein , was also expressed in the testis - supporting cells and interstitial cells . Our study showed that the expression of Annexin5 was decreased by 10.8 % in the acute stress group compared with the control group , and the expression of Annexin5 was decreased by 10.8 % in the Western blotting analysis .
Compared with the control group , the expression of Annexin5 mRNA was decreased by 17.4 % . The expression of Annexin5 mRNA was decreased by 17.4 % . The expression of Annexin5 mRNA was lower in the stress group than in the control group ( P < 0.05 ) .

The expression of Caspase - 3 mRNA and Bax / Bcl - 2 mRNA in rat testis were detected by RT - PCR . Compared with control group , there was no significant difference in the expression level of Bax / Bcl - 2 mRNA in acute stress group ( P > 0.05 ) .

Fear stress , as a form of psychological stress , has great damage and interference to the structure and function of rodent testis and reproductive endocrine , and promotes apoptosis of spermatogenic cells , suggesting that fear stress may have similar effects on human reproduction . Therefore , how to reduce fear stress injury is of great significance for maintaining normal reproduction function .

【学位授予单位】：南京师范大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R363

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