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

1.建立毛细管等速电泳（cITP）分析小鼠脂蛋白谱的方法，收集apoE缺陷小鼠合并血浆，用超离心法对血浆中各脂蛋白的比重进行标定，用cITP分析离心后血浆中脂蛋白的详细谱。结果，我们发现峰1-3、峰4、峰5和峰6-8分别对应于HDL、CM/VLDL、VLDL/IDL和VLDL/LDL。因此，cITP法可用于小鼠血浆脂蛋白谱的分析.重组apoE对apoE缺陷小鼠血浆脂蛋白谱的影响比较我们将apoE基因重组的apoE-2、apoE-3、apoE-4和apoE-仙台重组腺病毒载体分别导入apoE缺陷小鼠体内，通过cITP和肾组织学检测其血浆脂蛋白谱。在脂蛋白分析中，ApoE-Sendai引入的小鼠显示峰6升高，而其他apoE引入的小鼠显示峰7升高。在肾组织学上，76.5%的ApoE-仙台导入小鼠表现出类似脂蛋白肾小球病（LPG）的变化，而其他小鼠则表现出少许变化。（AN0VA_p<0.001）3. ApoE-仙台导入小鼠中LPG样改变的严重程度ApoE-仙台导入小鼠的组织学结果分为三个等级：轻度（46.1%）、中度（25.0%）和重度（5.8%）。轻度组仅表现为系膜脂蛋白沉积和空泡化，中度组可见少量脂蛋白血栓，重度组可见大量脂蛋白血栓。在一些肾小球中，系膜基质中的脂蛋白沉积物将迁移到毛细血管中。提示随着液化石油气的进展，脂蛋白可能从系膜基质转移到毛细血管腔。这些结果表明，cITP方法可用于分析小鼠血浆脂蛋白谱，并且电泳异常的VLDL/LDL（以峰6为界）与液化石油气的发生有关。

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.