尾悬吊模拟失重对大鼠卵巢功能及其组织中HIF-1α表达的影响

发布时间：2018-04-29 15:42

本文选题：模拟失重 + 卵巢　；参考：《安徽医科大学》2012年硕士论文

【摘要】：人类进入太空后，重力几乎完全消失，处于重力变化、辐射、噪音、昼夜节律丢失等各种复杂因素共存的空间环境中。这种复杂的环境会对人体心血管、肌肉、骨骼及血液系统造成很大影响。失重对生殖系统尤其是卵巢影响的研究报道较少。随着越来越多的女性航天员参与太空活动，研究失重对女性生殖系统的影响就显得尤为重要。本实验采用Morey-Holton尾悬吊模型来模拟失重环境，研究失重对雌性大鼠卵巢形态和功能以及卵巢组织中缺氧诱导因子-1α（hypoxia-induciblefactor-1alpha，HIF-1α）表达的影响。经医院伦理委员会批准后进行动物实验。主要研究内容和结果包括以下三部分。 1.采用尾部悬吊模拟失重，将25只成年健康雌性Wistar大鼠分为悬吊0d（地面对照组）、10d、20d、30d组和悬吊30d后解除悬吊恢复重力10d组，观察各实验组大鼠体重及生殖器官重量变化情况；HE染色观察卵巢卵泡变化情况；免疫组化方法观察卵泡增殖、凋亡情况；放射性免疫法（RIA）测定血清雌二醇、孕酮、FSH、LH含量。结果发现，悬吊组大鼠在悬吊5d后体重明显低于地面对照组，之后体重缓慢升高，解除悬吊恢复重力后，体重又明显下降；悬吊10d后，大鼠卵泡数量明显下降（P＜0.05），解除悬吊后恢复不明显；悬吊组大鼠与地面对照组相比，卵巢颗粒细胞增殖明显下降（P＜0.05），解除悬吊后有所恢复（P＜0.05）；悬吊组颗粒细胞凋亡明显增加（P＜0.05），恢复重力组有所下降，下降幅度不明显；RIA结果显示悬吊组大鼠E2含量明显下降（P＜0.05），LH、FSH含量升高（P＜0.05）。 2.选取15只成年健康雌性Wistar大鼠，随机分为地面对照组、悬吊30d组、悬吊30d后解除悬吊恢复重力10d组大鼠各5只，分别与健康成年Wistar雄性大鼠按1:1合笼，检测大鼠生殖能力。结果发现，与对照组相比，尾部悬吊30d大鼠产仔数明显降低（P＜0.05），恢复重力10d组大鼠产仔数有所增加（P＜0.05）。各实验组仔鼠出生体重无明显差异。另外，悬吊30d组大鼠，，有一只大鼠见拴后16d，体重停止增加且开始下降，最终未生育。 3.选取60只成年健康雌性Wistar大鼠，随机分为尾悬吊6h、12h、24h、2d、3d、5d、7d、14d、21d组和0h组（地面对照组），采用免疫组织化学和RT-PCR技术检测大鼠卵巢组织中HIF-1α的表达变化。结果发现，模拟失重6h、12h大鼠卵巢组织中HIF-1α蛋白和HIF-1α mRNA表达均明显高于对照组（P＜0.05）。综上所述，本研究运用常规石蜡染色、免疫组化、RT-PCR和RIA检测技术，系统观察和分析了模拟失重状态下大鼠卵巢形态和功能以及HIF-1α表达的变化。结果显示，模拟失重造成实验大鼠体重减轻，卵泡发育受到抑制，性激素分泌功能紊乱，卵巢储备功能和生育能力下降，这种影响通过重力的恢复而有一定程度的恢复。研究结果还提示，HIF-1α在模拟失重早期参与了大鼠卵巢的应激反应，并可能介导下游靶基因产生一系列保护效应。本研究提示，尾悬吊模拟失重对实验大鼠卵巢可造成一系列的应激性病理改变。卵巢对失重刺激在一定时限内可产生耐受乃至可能存在交叉耐受（cross-tolerance）以及解除悬吊后不同程度逆转恢复的现象和机制，均有待进一步研究。本研究为女性航天员在失重训练和航天活动中的保健防护提供了一定的参考依据。
[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.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R85

【参考文献】