Development of a novel method for analyzing murine lipoprotein profile and application for an experimental model of lipoprotein glomerulopathy

1.毛细管等速电泳法(CITP)分析小鼠脂蛋白谱方法的建立：采用超速离心法测定apoE缺陷小鼠的混合血浆中各脂蛋白的比重，并用CITP法测定离心后的血浆脂蛋白谱。结果表明，峰1-3、峰4、峰5和峰6-8分别对应于高密度脂蛋白、CM/极低密度脂蛋白、极低密度脂蛋白/低密度脂蛋白和极低密度脂蛋白/低密度脂蛋白。因此，CITP方法对于分析小鼠血浆脂蛋白谱是有用的。将载脂蛋白E2、E3、E4和ApoE-Sendai基因导入载脂蛋白E缺陷小鼠体内，用CITP法和肾脏组织学方法检测其血浆脂蛋白谱。在脂蛋白分析中，ApoE-仙台转基因小鼠的峰值6升高，而其他载脂蛋白转基因小鼠的峰值7升高。肾组织学观察，76.5%的ApoE-仙台转基因小鼠出现类似脂蛋白肾小球病变(LPG)的改变，而其他小鼠几乎没有变化。(Anova_p&lt；0.001)3.ApoE-仙台导入小鼠的液化石油气样变的严重程度。将ApoE-仙台导入小鼠的组织学结果分为三级：轻度(46.1%)，中度(25.0%)，重度(5.8%)。轻度组仅见系膜脂蛋白沉积和空泡化；中度组有少量脂蛋白血栓，重度组有大量脂蛋白血栓。)在几个肾小球中，系膜基质中的脂蛋白沉积将迁移到毛细血管中。提示随着LPG的进展，脂蛋白可能从系膜基质向毛细血管腔内迁移。提示CITP法可用于分析小鼠血浆脂蛋白谱，而以峰6为界的VLDL/LDL异常与LPG的发生发展有关。

1. Establishment of capillary isotachophoresis (cITP) method for murine lipoprotein profile analysis.Pooled plasma which was collected from apoE deficiency mice was demarcated by ultracentrifugation in specific gravity of each lipoprotein, and detailed lipoprotein profiles of centrifuged plasma were measured by cITP. As a result, we revealed that peak 1-3, peak 4, peak 5, and peak 6-8 corresponded to HDL, CM/VLDL, VLDL/IDL, and VLDL/LDL respectively. Therefore, It is demonstrated that cITP method is useful to analyze murine plasma lipoprotein profiles.2. A comparison of lipoprotein profile by cITP and histology of apoE deficiency mice administered by recombinant apoEs.We introduced apos E2, E3, and E4, and ApoE-Sendai inducted adenovirus vector to apoE deficiency mice, and examined their plasma lipoprotein profile by cITP and renal histology. In lipoprotein analysis, mice introduced by ApoE-Sendai showed rise of peak 6, whereas mice by other apoEs showed rise of peak 7. In renal histology, changes similar to lipoprotein glomerulopathy (LPG) were demonstrated in 76.5% of ApoE-Sendai introduced-mice, while a little change was appeared in other mice. (ANOVA_p<0.001)3. Severity of LPG like change in ApoE-Sendai introduced mice.Histlogical findings of ApoE-Sendai introduced mice were categorized into three grades; mild (46.1%), moderate (25.0%), and severe (5.8%). Mild group displayed only mesangial lipoprotein deposition and vacuolation; moderate group, a few lipoprotein thrombus, and severe group, many lipoprotein thrombi.) In several glomeruli, lipoprotein deposits in mesangial matrix were going to migrate into the capillary. It is suggested that lipoprotein may move from mesangial matrix to capillary lumen according to the progression of LPG.These results revealed that cITP method was available for analysis of murine plasma lipoprotein profile, and that abnormal electrophoretic VLDL/LDL, which is demarcated peak 6, is related to the development of LPG.