【作者】 张鹏飞；

【导师】 孙晓东；

【作者基本信息】 上海交通大学 ， 眼科学， 2016， 博士

【摘要】 目的:脉络膜新生血管(CNV)是湿性年龄相关性黄斑变性(AMD)的主要病理特征,M2型巨噬细胞极化参与了CNV的形成。因此,靶向调控M2型巨噬细胞极化有望成为干预CNV形成的新途径。本课题拟通过建立激光诱导小鼠CNV模型和原代小鼠骨髓巨噬细胞极化模型,了解巨噬细胞极化机制,探讨通过调控巨噬细胞极化干预CNV形成的效果,以期为治疗CNV提供新的思路。方法:(1)建立并评估激光诱导的小鼠CNV模型,采用western blotting、real time PCR和ELISA方法检测正常组织和激光光凝后RPE/脉络膜组织中巨噬细胞极化状况。(2)通过体外PGE2诱导原代小鼠骨髓巨噬细胞极化模型和激光诱导小鼠CNV模型,采用western blotting和real time PCR方法检测mi R-155通过调控C/EBPβ在巨噬细胞极化中的作用。(3)建立激光诱导小鼠CNV模型,玻璃体注射mi R-155 mimic,通过眼底荧光血管造影(FFA)、苏木精-伊红(HE)染色、脉络膜铺片和频域光相干断层扫描技术(SD-OCT)检测其对CNV形成的影响。结果:(1)M2型巨噬细胞极化指标Arg-1和Ym-1在激光诱导小鼠CNV模型组织中第1、3天高表达,差异有统计学意义(P<0.001),M1型巨噬细胞极化指标i NOS和Cox-2表达变化不明显。激光诱导小鼠CNV形成的第1、3天C/EBPβ和PGE2表达明显升高(P<0.001)。激光后第1、2、3天,局部组织中mi R-155表达显著降低,差异有统计学意义(P<0.001)。(2)500ng浓度PGE2干预原代小鼠骨髓巨噬细胞24小时后,M2型巨噬细胞极化指标Arg-1、Ym-1高表达,C/EBPβ表达升高,mi R-155表达下降(P<0.001),M1型巨噬细胞极化指标i NOS和Cox-2表达变化不明显。细胞转染mi R-155 mimic后再予PGE2干预可以明显减少Arg-1、Ym-1和C/EBPβ的表达(P<0.001)。(3)mi R-155 mimic玻璃体注射可以明显抑制第7、14天小鼠CNV形成(P<0.001)。结论:(1)小鼠CNV形成早期巨噬细胞发生M2型极化,PGE2是其重要刺激因素。(2)mi R-155可以通过调控C/EBPβ调节巨噬细胞极化。(3)mi R-155可以通过调节巨噬细胞M2型极化,抑制CNV的生长。目的:定量分析特发性黄斑裂孔(idiopathic macular hole,IMH)患者黄斑区脉络膜厚度改变。方法:采用横断面研究及meta分析。连续的IMH患者32例纳入研究,IMH患者散瞳后前置镜眼底检查,光相干断层扫描技术(OCT)检查确诊。选择同期年龄、性别、屈光度及眼轴匹配的正常人32名32只眼作为正常对照组。用加强成像深度扫描OCT(EDI-OCT)检测并比较IMH组及对照组黄斑中心凹脉络膜厚度(SFCT)和距离黄斑中心凹1mm、3mm上方、下方、鼻侧、颞侧脉络膜厚度。单因素和多因素分析黄斑区脉络膜厚度与各临床资料之间的关系。Meta分析用Stata软件计算两组之间的加权均数差。结果:IMH患者的平均SFCT为202.78±58.90μm,正常对照组为272.38±57.27μm。IMH组黄斑区各部位脉络膜厚度较对照组明显变薄,差异有统计学意义(P<0.001)。单因素与多因素线性回归分析,黄斑区脉络膜厚度与IMH诊断显著相关。Meta分析结果:IMH组的黄斑中央凹下脉络膜较正常组薄,加权平均差异为-56.98μm(95%CI:-68.56,-45.41)。结论:IMH患者黄斑区脉络膜厚度较正常眼薄,变薄的黄斑区脉络膜厚度与IMH诊断存在相关性。更多还原

【Abstract】 Purpose.M2-type macrophages play a significant role in the development of choroidal neovascularization(CNV).Thus,inhibition of M2 macrophage polarization may provide an alternative to anti-vascular endothelial growth factor(VEGF)therapy.This study evaluated the role of micro RNA-155(mi R-155)in M2 macrophage polarization,and assessed whether overexpression of mi R-155 could inhibit polarization of M2-type macrophages via targeting C/EBPβ and suppress choroidal neovascularization in laser-induced mice CNV model.Methods.(1)M2 macrophage polarization was evaluated in mice CNV model by western blotting,real time PCR and ELISA.(2)The effect of mi R-155 on M2 macrophage polarization was evaluated in mice CNV model and bone marrow-derived macrophages by western blotting,real time PCR.(3)To evaluate the role of mi R-155 on CNV development,eyes of mice in the mice CNV model were examined by Fluorescein Angiography(FA),hematoxylin and eosin(HE)staining,choroidal flatmounts and SD-OCT on days 7 and 14 after intravitreal injection.Results.(1)Arg–1 and Ym-1 were high expressed at days 1 and 3 in laser induced mice(P < 0.001),while i NOS and Cox-2 did not changed significantly.The expression of C/EBPβ and PGE2 were increased significantly at days 1 and 3(P < 0.001),while mi R-155 expression in local tissue decreased significantly at days 1,2,3 after laser(P < 0.001).(2)The expression of Arg-1,Ym-1 and C/EBPβ were high expressed in M2 bone marrow-derived macrophages cultured with PGE2 in vitro(P < 0.001),while mi R-155 was significantly decreased(P < 0.001),i NOS and Cox-2 did not changed obviously.(3)Intravitreal injection of mi R-155 mimic could inhibit CNV leakage and neovascularization at days 7 and 14 after laser in mice model(P < 0.001).Conclusion.Mi R-155 modulating C/EBPβ plays a significant role in M2 macrophage polarizaiton,while mi R-155 mimic could suppress choroidal neovascularization by inhibiting M2 macrophages polarization.Purpose: To investigate the choroidal thickness in unilateral idiopathic macular hole(IMH)eyes and compare them with normal control eyes using enhanced depth imaging optical coherence tomography(EDI-OCT).Methods: In this cross-sectional study,the subfoveal choroidal thickness(SFCT)and choroidal thickness at 1 mm and 3 mm nasal,temporal,superior,and inferior to the fovea of IMH eyes and normal control eyes were measured using EDI-OCT.Univariate and multivariate linear regression analyses were performed to evaluate the correlation between choroidal thicknesses at various locations and clinical factors.A meta-analysis was conducted using the Stata software package to calculate the summary of weighted mean differences(WMDs).Results: Thirty-two unilateral IMH patients and 32 controls were involved in this study.The IMH eyes had a thinner choroid than the control eyes at all macular locations(all P < 0.001).Multivariable linear regression analysis further showed that the choroidal thickness at any of the nine points was significantly thinner in association with the IMH diagnosis,as well as being somewhat thinner in association with the age and axial length.The result of our cross-sectional study is consistent with those of the meta-analysis with a pooled WMD of-56.99(95% confidence interval [CI]:-68.58 to-45.41)for SFCT.Conclusions: The study of Chinese unliateral IMH patients,along with the comprehensive meta-analysis,suggested that the choroidal thickness at all macular locations in unilateral IMH eyes was significantly decreased compared to the control group.更多还原