Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599, Singapore.
Khoo Teck Puat-National University Children's Medical Institute, National University Health System, Singapore 119260, Singapore.
Clin Sci (Lond). 2020 Jan 31;134(2):225-237. doi: 10.1042/CS20190961.
Circulating factors have been implicated in the pathogenesis of minimal change disease (MCD), and may have direct effects on cholesterol metabolism. This study investigated the pathogenesis of hypercholesterolemia in an IL-13 overexpression rat model of MCD prior to the onset of proteinuria, so as to establish the direct contribution of IL-13, especially with regard to hepatic cholesterol handling. In this model of MCD, the temporal relationship between hypercholesterolemia and proteinuria was first identified. Plasma proprotein convertase subtilisin/kexin type 9 (Pcsk9) and liver ATP-binding cassette sub-family G member 5 (Abcg5) were measured using ELISA. Liver Ldlr and liver X receptor alpha (Lxra) were quantified with Western blot. Abcg5-mediated cholesterol efflux in IL-13-stimulated rat primary hepatocytes was measured using taurocholate as cholesterol acceptor. The role of Lxra was validated using a luciferase assay in Lxre-luciferase-transfected IL-13-stimulated hepatocytes. IL-13-transfected rats developed hypercholesterolemia prior to proteinuria, with 35% of rats hypercholesterolemic but only 11% proteinuric by Day 20 (P = 0.04). These pre-proteinuric hypercholesterolemic rats showed elevations in total and LDL-cholesterol, but not hypertriglyceridemia or hepatic steatosis. The hypercholesterolemia was associated with increased hepatic Pcsk9 synthesis and enhanced circulating Pcsk9 levels, which correlated strongly with plasma total cholesterol (r = 0.73, P<0.001). The hypercholesterolemia was also contributed by decreased Abcg5 expression and activity, due to reduced Lxra expression. Lxra expression correlated with plasma total cholesterol levels (r = -0.52, P = 0.01), and overexpression of pLxra in rat hepatocytes abrogated the IL-13-mediated down-regulation of Lxre-driven gene expression. In conclusion, we have shown that IL-13 induced changes in hepatic cholesterol handling in a cytokine-induced rat model of MCD, resulting in hypercholesterolemia which can precede the onset of proteinuria.
循环因子与微小病变病(MCD)的发病机制有关,并且可能对胆固醇代谢有直接影响。本研究在蛋白尿发生之前,研究了 MCD 中白细胞介素 13(IL-13)过表达大鼠模型中高胆固醇血症的发病机制,从而确定了 IL-13 的直接作用,特别是在肝胆固醇处理方面。在这种 MCD 模型中,首先确定了高胆固醇血症与蛋白尿之间的时间关系。使用 ELISA 测量血浆前蛋白转化酶枯草溶菌素/克氏蛋白酶 9(Pcsk9)和肝 ATP 结合盒亚家族 G 成员 5(Abcg5)。使用 Western blot 定量肝 LDL 受体(Ldlr)和肝 X 受体α(Lxra)。使用牛磺胆酸盐作为胆固醇受体测量 IL-13 刺激的大鼠原代肝细胞中 Abcg5 介导的胆固醇流出。使用荧光素酶测定法在 Lxre-荧光素酶转染的 IL-13 刺激的肝细胞中验证 Lxra 的作用。转染 IL-13 的大鼠在蛋白尿之前就出现高胆固醇血症,20 天时 35%的大鼠高胆固醇血症,但仅有 11%的大鼠蛋白尿(P = 0.04)。这些预蛋白尿高胆固醇血症大鼠的总胆固醇和 LDL 胆固醇升高,但三酰甘油血症或肝脂肪变性不升高。高胆固醇血症与肝 Pcsk9 合成增加和循环 Pcsk9 水平升高有关,这与血浆总胆固醇密切相关(r = 0.73,P<0.001)。高胆固醇血症还归因于 Abcg5 表达和活性降低,这是由于 Lxra 表达降低所致。Lxra 表达与血浆总胆固醇水平相关(r = -0.52,P = 0.01),并且大鼠肝细胞中 pLxra 的过表达可消除 IL-13 介导的 Lxre 驱动基因表达的下调。总之,我们已经表明,在细胞因子诱导的 MCD 大鼠模型中,IL-13 诱导了肝胆固醇处理的变化,导致高胆固醇血症,这可能先于蛋白尿的发生。