摘要
目的 探索载脂蛋白E（ApoE）缺乏对视神经脊髓炎（NMO）体外模型的损伤影响。方法 将7 d龄的野生型及ApoE基因敲除（ApoE－/－）的C57BL/6J小鼠脊髓切片培养7 d后，将两种基因型的脊髓片按照随机数字表法随机分为实验组及对照组。实验组加入从NMO患者血清提取出的自身抗体免疫球蛋白G（NMO－IgG）及正常人补体血清；对照组不加NMO－IgG，余处理方式与实验组相同，继续培养24 h。用免疫荧光法检测脊髓片水通道蛋白4（AQP4）、胶质纤维酸性蛋白（GFAP）、离子钙接头蛋白（IBA1）、髓鞘碱性蛋白（MBP）、人神经丝蛋白L（NFL）的表达水平，并根据脊髓片AQP4和GFAP缺失面积比例计算损伤积分。结果 与相应的对照组相比，ApoE基因敲除小鼠造模组（NMO－ApoE－/－组）和野生型小鼠造模组（NMO－WT组）均出现明显的AQP4、GFAP、MBP和NFL缺失（NMO－ApoE－/－组上述各项指标缺失面积百分比分别为83.88%±5.01%、82.44%±6.11%、45.02%±5.11%、54.65%±7.66%，相应对照组对应指标分别为10.44%±4.07%、5.73%±0.82%、9.12%±1.41%、5.72%±0.81%, t＝34.143、37.269、20.300、19.051，均P〈0.05；NMO－WT组上述指标缺失面积比例分别为77.74%±6.75%、75.62%±5.76%、37.60%±4.88%、46.29%±4.98%，相应对照组对应指标分别为9.31%±2.97%、5.80%±0.82%、9.10%±1.63%、5.80%±0.81%，t＝27.828、35.934、16.613、24.057，均P〈0.05），且NMO－ApoE－/－组染色缺失的程度较NMO－WT组严重（t＝2.194、2.436、3.149、2.746，均P〈0.05），两组损伤积分均较各自对照组明显增高，均出现IBA1表达上调（NMO－WT组及对照组：19.88±1.11与11.18±0.65，t＝25.270，P〈0.05），且NMO－ApoE－/－组的IBA1表达水平较NMO－WT组更高（25.81±1.61与19.88±1.11，t＝9.101，P〈0.05）。结论 在补体存在的情况下，从NMO患者血清提取出的NMO－IgG可以诱导离体组织产生NMO样损伤。ApoE缺失可�
Objective To investigate the effects of apolipoprotein E（ApoE） deficiency on neuromyelitis optica （NMO） model of spinal cord sections induced by NMO-IgG and complement in vitro.Methods NMO-IgG was extracted from the patients with NMO, and complementary serum from healthy people. The spinal cord sections of seven days old C57BL / 6J mice with wild type （WT） or ApoE knockout （ApoE-/-） were cultured for seven days. The spinal cords of the two genotypes were randomly divided into experimental groups （NMO-ApoE-/- group, NMO-WT group） and control groups （C-AopE-/- group, C-WT group）, respectively. The experimental groups were treated with NMO-IgG and complementary serum, and the control groups only with complementary serum. Then all the sections were continued incubating for 24 h before harvested. Immunofluorescence staining and modified thick tissue film immunofluorescence were used to detect the expression of aquaporin 4 （AQP4）, glial fibrillary acidic protein （GFAP）, ionic calcium fibronectin （IBA1）, myelin basic protein （MBP） and human neurofilament protein L （NFL） respectively. The lesion score was calculated according to the areas percentage of AQP4 and GFAP deficiency in spinal cord sections.Results Compared with the respective control groups, the expressions of AQP4, GFAP, MBP and NFL were deficient in the experimental groups （The percentages of missing area in the NMO-ApoE-/- group were 83.88%±5.01%, 82.44%±6.11%, 45.02%±5.11% and 54.65%±7.66% respectively, while the percentages of missing area in the C-ApoE-/- group were 10.44%±4.07%, 5.73%±0.82%, 9.12%±1.41% and 5.72%±0.81%, t=34.143, 37.269, 20.300, 19.051, all P〈0.05; The percentages of missing area in the NMO-WT group were 77.74%±6.75%, 75.62%±5.76%, 37.60%±4.88% and 46.29%±4.98%, while the percentages of missing area in the C-WT group were 9.31%±2.97%, 5.80%±0.82%, 9.10%±1.63%, 5.80%±0.81% respectively, t=27.828, 35.934, 16.613, 24.057, all P〈0.05）. While IBA1 was up-regulated and the da
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2018年第2期111-117,共7页
Chinese Journal of Neurology
基金
国家自然科学基金资助项目（81460194,81260188）
广西自然科学基金资助项目（2015GXNSFAA139212）
