本文選題：模擬失重 + 卵巢�。� 參考：《安徽醫(yī)科大學(xué)》2012年碩士論文

【摘要】：人類進(jìn)入太空后，重力幾乎完全消失，處于重力變化、輻射、噪音、晝夜節(jié)律丟失等各種復(fù)雜因素共存的空間環(huán)境中。這種復(fù)雜的環(huán)境會(huì)對人體心血管、肌肉、骨骼及血液系統(tǒng)造成很大影響。失重對生殖系統(tǒng)尤其是卵巢影響的研究報(bào)道較少。隨著越來越多的女性航天員參與太空活動(dòng)，研究失重對女性生殖系統(tǒng)的影響就顯得尤為重要。本實(shí)驗(yàn)采用Morey-Holton尾懸吊模型來模擬失重環(huán)境，研究失重對雌性大鼠卵巢形態(tài)和功能以及卵巢組織中缺氧誘導(dǎo)因子-1α（hypoxia-induciblefactor-1alpha，HIF-1α）表達(dá)的影響。經(jīng)醫(yī)院倫理委員會(huì)批準(zhǔn)后進(jìn)行動(dòng)物實(shí)驗(yàn)。主要研究內(nèi)容和結(jié)果包括以下三部分。 1.采用尾部懸吊模擬失重，將25只成年健康雌性Wistar大鼠分為懸吊0d（地面對照組）、10d、20d、30d組和懸吊30d后解除懸吊恢復(fù)重力10d組，觀察各實(shí)驗(yàn)組大鼠體重及生殖器官重量變化情況；HE染色觀察卵巢卵泡變化情況；免疫組化方法觀察卵泡增殖、凋亡情況；放射性免疫法（RIA）測定血清雌二醇、孕酮、FSH、LH含量。結(jié)果發(fā)現(xiàn)，懸吊組大鼠在懸吊5d后體重明顯低于地面對照組，之后體重緩慢升高，解除懸吊恢復(fù)重力后，體重又明顯下降；懸吊10d后，大鼠卵泡數(shù)量明顯下降（P＜0.05），解除懸吊后恢復(fù)不明顯；懸吊組大鼠與地面對照組相比，卵巢顆粒細(xì)胞增殖明顯下降（P＜0.05），解除懸吊后有所恢復(fù)（P＜0.05）；懸吊組顆粒細(xì)胞凋亡明顯增加（P＜0.05），恢復(fù)重力組有所下降，下降幅度不明顯；RIA結(jié)果顯示懸吊組大鼠E2含量明顯下降（P＜0.05），LH、FSH含量升高（P＜0.05）。 2.選取15只成年健康雌性Wistar大鼠，隨機(jī)分為地面對照組、懸吊30d組、懸吊30d后解除懸吊恢復(fù)重力10d組大鼠各5只，分別與健康成年Wistar雄性大鼠按1:1合籠，檢測大鼠生殖能力。結(jié)果發(fā)現(xiàn)，與對照組相比，尾部懸吊30d大鼠產(chǎn)仔數(shù)明顯降低（P＜0.05），恢復(fù)重力10d組大鼠產(chǎn)仔數(shù)有所增加（P＜0.05）。各實(shí)驗(yàn)組仔鼠出生體重?zé)o明顯差異。另外，懸吊30d組大鼠，，有一只大鼠見拴后16d，體重停止增加且開始下降，最終未生育。 3.選取60只成年健康雌性Wistar大鼠，隨機(jī)分為尾懸吊6h、12h、24h、2d、3d、5d、7d、14d、21d組和0h組（地面對照組），采用免疫組織化學(xué)和RT-PCR技術(shù)檢測大鼠卵巢組織中HIF-1α的表達(dá)變化。結(jié)果發(fā)現(xiàn)，模擬失重6h、12h大鼠卵巢組織中HIF-1α蛋白和HIF-1α mRNA表達(dá)均明顯高于對照組（P＜0.05）。 綜上所述，本研究運(yùn)用常規(guī)石蠟染色、免疫組化、RT-PCR和RIA檢測技術(shù)，系統(tǒng)觀察和分析了模擬失重狀態(tài)下大鼠卵巢形態(tài)和功能以及HIF-1α表達(dá)的變化。結(jié)果顯示，模擬失重造成實(shí)驗(yàn)大鼠體重減輕，卵泡發(fā)育受到抑制，性激素分泌功能紊亂，卵巢儲(chǔ)備功能和生育能力下降，這種影響通過重力的恢復(fù)而有一定程度的恢復(fù)。研究結(jié)果還提示，HIF-1α在模擬失重早期參與了大鼠卵巢的應(yīng)激反應(yīng)，并可能介導(dǎo)下游靶基因產(chǎn)生一系列保護(hù)效應(yīng)。本研究提示，尾懸吊模擬失重對實(shí)驗(yàn)大鼠卵巢可造成一系列的應(yīng)激性病理改變。卵巢對失重刺激在一定時(shí)限內(nèi)可產(chǎn)生耐受乃至可能存在交叉耐受（cross-tolerance）以及解除懸吊后不同程度逆轉(zhuǎn)恢復(fù)的現(xiàn)象和機(jī)制，均有待進(jìn)一步研究。本研究為女性航天員在失重訓(xùn)練和航天活動(dòng)中的保健防護(hù)提供了一定的參考依據(jù)。
[Abstract]:When human beings enter space, gravity almost completely disappears, in a space environment where complex factors coexist in a variety of complex factors, such as gravity change, radiation, noise, and circadian rhythm loss. This complex environment has a great impact on human cardiovascular, muscle, bone and blood systems. As more and more female astronauts participate in space activities, it is very important to study the effect of weightlessness on the female reproductive system. In this experiment, the Morey-Holton tail suspension model was used to simulate the weightlessness environment and to study the morphology and function of the female rat ovary and the hypoxia inducible factor -1 alpha (hypoxia-induciblefactor-) in the ovarian tissue. The influence of 1alpha, HIF-1 alpha expression on animal experiment was approved by the hospital ethics committee. The main research contents and results included the following three parts.
1. using tail suspension to simulate weightlessness, 25 adult healthy female Wistar rats were divided into suspended 0d (ground control group), 10d, 20d, 30d group and suspended 30d were removed to restore gravity 10d group, and the weight and reproductive organs weight of the rats in the experimental groups were observed. The changes of ovarian follicles were observed by HE staining, and the eggs were observed by immunohistochemical method. Proliferation, apoptosis, serum estradiol, progesterone, FSH and LH in serum were measured by radioimmunoassay (RIA). The results showed that the weight of rats in suspension group was significantly lower than that of the ground control group after suspended 5D. After the suspension of gravity, the weight of the rats decreased obviously; after suspended 10D, the number of follicles in the rats decreased significantly (P < 0.05). In suspension group, the proliferation of ovarian granulosa cells decreased significantly (P < 0.05) compared with the ground control group (P < 0.05), and the apoptosis of granular cells in suspension group increased significantly (P < 0.05), the recovery of gravity group decreased and the decrease was not obvious; RIA results showed that the E2 contained in the suspension group was E2 The amount decreased significantly (P < 0.05), and LH and FSH contents increased (P < 0.05).
2. 15 adult healthy female Wistar rats were randomly divided into the ground control group and the suspended 30d group. After suspended 30d, 5 rats were suspended to restore the gravity 10d group. The reproductive ability of the rats was detected by 1:1 cage with the healthy adult Wistar male rats respectively. The results showed that the litter size of the tail suspended 30d rats was significantly lower than the control group (P < 0.). 05), the litter size of rats in the 10d group was increased (P < 0.05). There was no significant difference in birth weight of the experimental group. In addition, the rats in the suspension group of 30d group, one of the rats, saw the post tethered 16d, and the weight stopped and began to decline and finally had no birth.
3. 60 adult healthy female Wistar rats were randomly divided into tail suspension 6h, 12h, 24h, 2D, 3D, 5D, 7d, 14d, 21d group and 0h group (ground control group). The expression of alpha protein in the ovarian tissue of rats was detected by immunohistochemistry and RT-PCR technology. All of them were significantly higher than those of the control group (P < 0.05).
To sum up, this study used routine paraffin staining, immunohistochemistry, RT-PCR and RIA detection techniques to systematically observe and analyze the changes in the morphology and function of the ovaries and the expression of HIF-1 alpha in the rats under simulated weightlessness. The results showed that simulated weightlessness resulted in the loss of weight, the inhibition of follicle development, the disorder of the sex hormone secretion, and the disorder of the sex hormone secretion. The results also suggest that HIF-1 alpha participates in the stress response of the rat ovaries in the early stage of simulated weightlessness and may mediate a series of protective effects on the downstream target genes. The nests can cause a series of stressful pathological changes. The phenomenon and mechanism of tolerance and possible existence of cross tolerance (cross-tolerance) and reversion recovery after lifting suspension in a certain time limit for weightlessness stimulation are still needed to be further studied. This study is for female astronauts in weightlessness training and space activities. Health protection provides a certain reference basis.

【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R85

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