【摘要】： 研究背景/目的目前近視作為全球發(fā)生率最高的一種屈光不正,因發(fā)病率高、危害性大已經(jīng)成為全球防治的重點(diǎn),其防治工作已被世界衛(wèi)生組織(WHO)列入防盲計(jì)劃。 關(guān)于近視的研究歷史已有200余年,就近視的發(fā)生機(jī)制人們也曾提出許多假說,但至今尚不能明確,更沒有確切有效的防治方法。多項(xiàng)實(shí)驗(yàn)研究認(rèn)為,形覺剝奪性近視的發(fā)生可能是由局部視網(wǎng)膜調(diào)控機(jī)制決定的。 近幾年研究發(fā)現(xiàn),血管活性腸肽(vasoactive intestinal peptide, VIP)與近視有著密切的相關(guān)性。作為視覺信息分子,VIP在視覺誘導(dǎo)近視眼形成的過程中可能起到視覺信息傳遞和調(diào)控的作用。在幼雞及靈長(zhǎng)類等近視動(dòng)物模型中發(fā)現(xiàn)VIP在視網(wǎng)膜中的表達(dá)明顯增高,并在幼雞近視動(dòng)物模型中隨近視程度的增加表達(dá)增加,但在更接近人的哺乳動(dòng)物形覺剝奪性近視(form deprivation myopia, FDM)模型中是否也存在同樣的動(dòng)態(tài)變化尚未得到證實(shí)。 本研究擬用不同技術(shù)方法,觀察并對(duì)比VIP在豚鼠近視模型中形覺剝奪不同時(shí)間后實(shí)驗(yàn)組與對(duì)照組視網(wǎng)膜中的表達(dá)情況,同時(shí)觀察其變化趨勢(shì)是否與在幼雞中的變化一致,進(jìn)一步了解其在哺乳動(dòng)物中可能參與的發(fā)生或發(fā)展機(jī)制,為尋求一種新的治療或預(yù)防方法提供理論基礎(chǔ)。 方法選取60只2-3周齡三色健康豚鼠,隨機(jī)選40只隨機(jī)分成遮蓋前、遮蓋1w、2w、3w組,每組10只,采用單眼遮蓋法(monocular deprivation, MD)進(jìn)行形覺剝奪,均以右眼為剝奪眼(實(shí)驗(yàn)組),左眼暴露為自身對(duì)照組。另外分別在相應(yīng)時(shí)間點(diǎn)隨機(jī)選取5只同齡、未遮蓋豚鼠作為正常對(duì)照組。 在遮蓋前和MD 1w、2w、3w后對(duì)相應(yīng)豚鼠進(jìn)行屈光度和眼軸長(zhǎng)度的測(cè)量。遮蓋組、自身對(duì)照組、正常對(duì)照組各隨機(jī)取5只豚鼠的眼球用于HE染色和免疫組織化學(xué)；另5只豚鼠取視網(wǎng)膜組織,用于RT-PCR。對(duì)結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果①形覺剝奪前各組間屈光度和眼軸長(zhǎng)度在統(tǒng)計(jì)學(xué)無明顯差異(P0.05),在單眼形覺剝奪后的1w、2w、3w,各組之間的差異在統(tǒng)計(jì)學(xué)上均具有顯著性(屈光度P0.01、P0.01、P0.05,眼軸長(zhǎng)度均P0.01)；實(shí)驗(yàn)組、自身對(duì)照組屈光度和眼軸長(zhǎng)度隨形覺剝奪時(shí)間的增加而增加,正常對(duì)照組則在統(tǒng)計(jì)學(xué)上無顯著性差異(P0.05)。②形覺剝奪前各組間視網(wǎng)膜VIP在蛋白及mRNA水平無明顯統(tǒng)計(jì)學(xué)差異(P0.05),單眼形覺剝奪后的1w、2w、3w,各組之間的差異在統(tǒng)計(jì)學(xué)上均具有顯著性(均P0.01)；實(shí)驗(yàn)組、自身對(duì)照組視網(wǎng)膜VIP蛋白、VIP mRNA隨形覺剝奪時(shí)間的增加而增加,正常對(duì)照組在統(tǒng)計(jì)學(xué)上無顯著性差異(P0.05)。 結(jié)論①形覺剝奪1w可以形成近視,且屈光度和眼軸長(zhǎng)度隨形覺剝奪時(shí)間的延長(zhǎng)而增加。②VIP可能與形覺剝奪性近視的發(fā)生或發(fā)展有關(guān)。VIP在FDM豚鼠視網(wǎng)膜的表達(dá)隨形覺剝奪時(shí)間的延長(zhǎng)而增加,與屈光度和眼軸長(zhǎng)度的變化一致。
[Abstract]:Background / objective myopia, as one of the most common refractive errors in the world, has become the focus of global prevention and control because of its high incidence. The prevention and treatment of myopia has been included in the World Health Organization (WHO) blind prevention program. There have been more than 200 years of research on myopia. Many hypotheses have been put forward on the mechanism of myopia, but so far it is not clear, let alone effective prevention and cure methods. Many experimental studies suggest that the occurrence of form-deprived myopia may be determined by the mechanism of local retinal regulation. In recent years, it has been found that vasoactive intestinal peptide (vasoactive intestinal peptide, VIP) is closely related to myopia. VIP, as a visual information molecule, may play a role in the transmission and regulation of visual information during the formation of myopia induced by vision. The expression of VIP was significantly increased in the retina of young chickens and primates, and increased with the increase of myopia. But the same dynamic changes have not been confirmed in the mammalian (form deprivation myopia, FDM) model of form deprivation myopia. This study was designed to observe and compare the expression of VIP in the retina of experimental group and control group after form-deprivation in guinea pig myopia model with different techniques, and to observe whether the change trend of VIP was consistent with that in young chicken. Further understanding the mechanism of its possible involvement in mammals provides a theoretical basis for seeking a new method of treatment or prevention. Methods A total of 60 healthy guinea pigs of 2-3 weeks old were randomly divided into two groups: one week and two weeks after 3 weeks of shading, 10 guinea pigs in each group were deprived of shape by monocular shading (monocular deprivation, MD). The right eye was taken as the deprivation eye (experimental group) and the left eye was exposed as the self-control group. In addition, 5 guinea pigs of the same age were randomly selected as the normal control group at the corresponding time points. The diopter and the length of the eye axis of the guinea pigs were measured before the occlusion and after 1 week and 2 weeks of MD. The eyeballs of 5 guinea pigs in shaded group, self control group and normal control group were randomly selected for HE staining and immunohistochemistry, and retinal tissues were taken from 5 guinea pigs for RT-PCR.. The results were analyzed statistically. Results there was no significant difference in diopter and eye axis length between the groups before and after the deprivation (P0.05), but there were significant differences in the diopter and the length of the eye axis between the three groups at 1 week and 2 weeks after the deprivation (P 0.01, P 0.01, P 0.01), while in the experimental group, there was no significant difference in the diopter and the length of the eye axis (P 0.01), while in the experimental group, there was no significant difference in the diopter and the length of the eye axis (P 0.01). The diopter and axial length of the control group increased with the increase of the deprivation time. In the normal control group, there was no significant difference in protein and mRNA levels in retina before and after deprivation of shape (P0.05), but there was no significant difference in protein and mRNA levels in the retina of the normal control group (P0.05). The difference was statistically significant between the three groups at 1 week and 2 ws after monocular deprivation (P0.05). There was significant difference (P0.01). In the experimental group, the retinal VIP mRNA increased with the increase of form deprivation time in the control group, but there was no significant difference in the normal control group (P0.05). Conclusion 1 shape deprivation for 1 week can cause myopia. The diopter and the length of eye axis increased with the prolongation of the deprivation time. 2VIPs may be related to the occurrence or development of formal-deprivation myopia. The expression of VIP in the retina of FDM guinea pigs increased with the prolongation of the deprivation time. It is consistent with diopter and eye axis length.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R778.11

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