本文選題：恐懼 + 應激��； 參考：《南京師范大學》2007年碩士論文

【摘要】： 應激(Stress)是指機體在受到各種內外環(huán)境因素刺激時所發(fā)生的非特異性全身反應。長期處于應激狀態(tài)，機體的自穩(wěn)態(tài)將改變，從而誘發(fā)機體產生疾病。遭受戰(zhàn)爭環(huán)境刺激的軍人喪失了應付能力，并伴有精神障礙，對此稱之為作戰(zhàn)應激反應(combat stress reaction CRS)；發(fā)展到后期會導致創(chuàng)傷后應激性障礙(post-traumatic stress disorder，PTSD)。 本實驗建立戰(zhàn)爭恐懼應激大鼠模型，檢測恐懼應激對生殖的影響。研究表明急性應激組與對照組相比，大鼠體重、睪丸重量、附睪重量、精子數量沒有明顯變化(P＞0.05)，睪丸HE染色顯示睪丸結構無明顯變化，免疫組化顯示3β-HSD表達降低，Western blotting分析3β-HSD降低了7％；慢性應激組與對照組相比，大鼠體重、睪丸重量、附睪重量、精子數量都有顯著差異，P＜0.05。睪丸HE染色顯示睪丸內生精小管細小，管腔內生精細胞稀少，免疫組化顯示3β-HSD表達降低，Western blotting分析3β-HSD降低了19.6％。因此表明隨著應激時間的延長生殖器官功能降低。 此外，我們用酶聯免疫吸附法檢測應激組與對照組大鼠血清FSH、LH和T水平，發(fā)現急性應激組大鼠血清FSH(2.7±0.4)ng／ml、LH(3.1±0.3)ng／ml和T(3.9±0.5)ng／ml水平略高于對照組大鼠FSH(2.6±0.2)ng／ml、LH(2.8±0.6)ng／ml和T(2.7±0.2)ng／ml水平，但無顯著差異(P＞0.05)；亞急性應激組大鼠血清FSH(1.9±0.5)ng／ml、LH(2.9±0.3)ng／ml和T(1.9±1.1)ng／ml水平都低于其對照組大鼠的FSH(2.3±0.4)ng／ml、LH(3.0±0.5)ng／ml和T(2.9±1.3)ng／ml水平，但都無顯著差異(P＞0.05)；慢性應激組大鼠血清FSH(1.6±0.5)ng／ml、LH(2.1±0.4)ng／ml和T(0.6±0.3)ng／ml都低于對照組大鼠血清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水平的變化有顯著差異(P＜0.05)；而T水平的變化有極顯著差異(P＜0.001)。提示恐懼應激會擾亂大鼠的正常生殖內分泌功能。 Annexin5一種鈣依賴的磷脂酶結合蛋白，在睪丸支持細胞和間質細胞中也有表達。我們研究表明急性應激組與對照組相比，免疫組化顯示Annexin5都定位在間質細胞、支持細胞和成熟的精子頭部，Western blotting分析發(fā)現Annexin5的表達降低了10.8％；慢性應激組與對照組相比，免疫組化結果顯示Annexin5在精子頭部的定位逐漸減少，應激1w和2w組，未見到Annexin5定位于精子頭部，到了應激的3w和4w，發(fā)現Annexin5重新定位于精子的頭部，而在間質細胞和支持細胞上的定位模式沒有顯著變化。Western blotting分析發(fā)現Annexin5的表達降低了17.4％。我們用RT-PCR檢測了Annexin5mRNA的表達。應激組與各自的對照組相比，Annxin5 mRNA的表達都低于對照組，其中亞急性組與對照組相比有顯著差異(P＜0.05)。隨著應激時間的延長，Annexin5 mRNA由降低逐漸向生理水平回升。這些數據表明，應激改變大鼠睪丸中Annexin5的正常表達模式。 最后，我們還用RT-PCR檢測了大鼠睪丸Caspase-3 mRNA和Bax／Bcl-2 mRNA表達的變化。急性應激組與對照組相比，Caspase-3 mRNA的表達水平無顯著變化(P＞0.05)，而亞急性應激組、慢性應激組與其對照組相比都有顯著差異(P＜0.05)。與對照組相比，急性應激組Bax／Bcl-2 mRNA比值無顯著變化(P＞0.05)，而亞急性應激組有顯著差異(P＜0.05)，慢性應激組有極顯著差異(P＜0.001)。TUNEL結果表明急性應激組與對照組相比，生殖細胞的凋亡指數無顯著差異(P＞0.05)，而亞急性應激組和慢性應激組與對照相比生殖細胞凋亡指數有極顯著差異(P＜0.001)。因此，長期恐懼應激可導致大鼠睪丸生精細胞發(fā)生凋亡，而這種凋亡有Caspase-3及bax和bcl-2等的參與。 恐懼應激作為心理應激的一種形式對嚙齒類動物睪丸的結構和功能以及生殖內分泌等都有著很大的破壞和干擾作用，并且促使睪丸生精細胞的凋亡，，提示恐懼應激對人類生殖可能有相似的影響。因此，如何減少恐懼應激損傷，對于維持正常生殖功能有著重要的意義。
[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 鹵 0.4 ) ng / ml , LH ( 3.1 鹵 0.3 ) ng / ml and T ( 3.9 鹵 0.5 ) ng / ml in acute stress group were slightly higher than those in control group ( 2.6 鹵 0.2 ) ng / ml , LH ( 2.8 鹵 0.6 ) ng / ml and T ( 2.7 鹵 0.2 ) ng / ml , but there was no significant difference ( P > 0.05 ) .
The levels of FSH ( 1.9 鹵 0.5 ) ng / ml , LH ( 2.9 鹵 0.3 ) ng / ml and T ( 1.9 鹵 1.1 ) ng / ml in subacute stress group were lower than those in control rats ( 2.3 鹵 0.4 ) ng / ml , LH ( 3.0 鹵 0.5 ) ng / ml and T ( 2.9 鹵 1.3 ) ng / ml , but there was no significant difference ( P > 0.05 ) .
The serum levels of FSH ( 1.6 鹵 0.5 ) ng / ml , LH ( 2.1 鹵 0.4 ) ng / ml and T ( 0.6 鹵 0.3 ) ng / ml in chronic stress group were lower than those in control group ( 2.2 鹵 0.3 ) ng / ml , LH ( 3.0 鹵 0.8 ) ng / ml and T ( 3.0 鹵 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 .

【學位授予單位】：南京師范大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R363

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