血管活性肠肽（VIP）在豚鼠形觉剥夺性近视眼中表达的动态观察

发布时间：2018-09-07 15:59

【摘要】： 研究背景/目的目前近视作为全球发生率最高的一种屈光不正,因发病率高、危害性大已经成为全球防治的重点,其防治工作已被世界卫生组织(WHO)列入防盲计划。关于近视的研究历史已有200余年,就近视的发生机制人们也曾提出许多假说,但至今尚不能明确,更没有确切有效的防治方法。多项实验研究认为,形觉剥夺性近视的发生可能是由局部视网膜调控机制决定的。近几年研究发现,血管活性肠肽(vasoactive intestinal peptide, VIP)与近视有着密切的相关性。作为视觉信息分子,VIP在视觉诱导近视眼形成的过程中可能起到视觉信息传递和调控的作用。在幼鸡及灵长类等近视动物模型中发现VIP在视网膜中的表达明显增高,并在幼鸡近视动物模型中随近视程度的增加表达增加,但在更接近人的哺乳动物形觉剥夺性近视(form deprivation myopia, FDM)模型中是否也存在同样的动态变化尚未得到证实。本研究拟用不同技术方法,观察并对比VIP在豚鼠近视模型中形觉剥夺不同时间后实验组与对照组视网膜中的表达情况,同时观察其变化趋势是否与在幼鸡中的变化一致,进一步了解其在哺乳动物中可能参与的发生或发展机制,为寻求一种新的治疗或预防方法提供理论基础。方法选取60只2-3周龄三色健康豚鼠,随机选40只随机分成遮盖前、遮盖1w、2w、3w组,每组10只,采用单眼遮盖法(monocular deprivation, MD)进行形觉剥夺,均以右眼为剥夺眼(实验组),左眼暴露为自身对照组。另外分别在相应时间点随机选取5只同龄、未遮盖豚鼠作为正常对照组。在遮盖前和MD 1w、2w、3w后对相应豚鼠进行屈光度和眼轴长度的测量。遮盖组、自身对照组、正常对照组各随机取5只豚鼠的眼球用于HE染色和免疫组织化学；另5只豚鼠取视网膜组织,用于RT-PCR。对结果进行统计学分析。结果①形觉剥夺前各组间屈光度和眼轴长度在统计学无明显差异(P0.05),在单眼形觉剥夺后的1w、2w、3w,各组之间的差异在统计学上均具有显著性(屈光度P0.01、P0.01、P0.05,眼轴长度均P0.01)；实验组、自身对照组屈光度和眼轴长度随形觉剥夺时间的增加而增加,正常对照组则在统计学上无显著性差异(P0.05)。②形觉剥夺前各组间视网膜VIP在蛋白及mRNA水平无明显统计学差异(P0.05),单眼形觉剥夺后的1w、2w、3w,各组之间的差异在统计学上均具有显著性(均P0.01)；实验组、自身对照组视网膜VIP蛋白、VIP mRNA随形觉剥夺时间的增加而增加,正常对照组在统计学上无显著性差异(P0.05)。结论①形觉剥夺1w可以形成近视,且屈光度和眼轴长度随形觉剥夺时间的延长而增加。②VIP可能与形觉剥夺性近视的发生或发展有关。VIP在FDM豚鼠视网膜的表达随形觉剥夺时间的延长而增加,与屈光度和眼轴长度的变化一致。
[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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R778.11

【相似文献】