ApoE基因多态性对瑞舒伐他汀治疗高脂血症疗效的影响及其机制的研究

目的：探讨ApoE基因多态性对瑞舒伐他汀（rosuvastatin，RSS）治疗高脂血症疗效的影响及其具体机制。方法：选取2018年1月1日至12月31日在郑州大学第二附属医院诊断的高血脂患者910例，采用"基因芯片法（Gene Chip）"分析ApoE基因型，并依据等位基因分组（ε2组、ε3组及ε4组），于RSS用药前及连续用药8周后采集静脉血，比较各组患者RSS用药前后总胆固醇（total cholesterol，TC）、三酰甘油（triglyceride，TG）、高密度脂蛋白胆固醇（high density liptein cholesterol，HDL-C）、低密度脂蛋白（low density lipoprotein，LDL）及脂蛋白a[lipoprotein-a，Lp （a）]变化水平，RT-PCR法检测用药前后ApoE mRNA表达；无血清培养正常人L-02型肝细胞诱导低密度脂蛋白受体（low density lipoprotein receptor，LDL-R）活性，分别加入不同浓度ApoE2/3/4-DMPC复合物，ELISA法比较ApoE2/3/4竞争性结合LDL-R的能力，RT-PCR法比较RSS用药后ApoE2/3/4对LDLR mRNA、HMG-CoA mRNA表达的影响。结果：ε2组（ε2/ε2、ε2/ε3）患者RSS治疗后TC、TG、LDL-C、Lp （a）下降水平，HDL-C升高水平显著高于ε3组（ε3/ε3、ε2/ε4）及ε4组（ε3/ε4、ε4/ε4）患者，其中ε4组疗效最差；ε2组（ε2/ε2、ε2/ε3）及ε3组（ε3/ε3）、ε4组（ε3/ε4） RSS用药后较用药前ApoE mRNA相对表达量显著降低（P<0.05），但ε3组（ε2/ε4）与ε4组（ε4/ε4）两种基因型RSS用药前后无显著性统计学差异（P>0.05）。在L-02肝细胞中，ApoE2与LDL-R结合力高于ApoE3（P<0.05），ApoE4与LDL-R结合力最弱；与ApoE3比较，ApoE2可显著上调LDL-R mRNA表达（P<0.05），但ApoE4则抑制LDL-R mRNA表达（P<0.05）；ApoE2抑制HMG-CoA mRNA表达的程度显著高于ApoE3（P<0.05），而ApoE4抑制HMG-CoA mRNA表达的程度显著低于ApoE3（P<0.05）。结论：RSS对于ApoE ε2等位基因患者降低血脂的疗效最优，其次为ε3、ε4等位基因携带者，该作用可能与ApoE2诱导LDLR基因表达，增强ApoE2与LDLR亲和力，从而增强RSS对HMG-CoA基因表达抑制作用有关。

Effect of ApoE gene polymorphism on the efficacy of rosuvastatin in the treatment of hyperlipidemia and its mechanism

OBJECTIVE To investigate the effect of ApoE gene polymorphism on the efficacy of rosuvastatin (RSS) in the treatment of hyperlipidemia and its specific mechanism.METHODS A total of 910 patients with hyperlipidemia who were diagnosed in the Second Affiliated Hospital of Zhengzhou University on January 1, 2018 were selected and analyzed for ApoE genotype by "Gene Chip". Venous blood was collected after 8 weeks of continuous treatment with RSS according to allele groups (ε2 group and ε3 group). The changes of total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein (LDL) and lipoprotein a [Lp (a)] before and after treatment with RSS were compared. The expression of ApoE mRNA was detected by RT-PCR. Normal L-02 hepatocytes were cultured without serum to induce low density lipoprotein receptor (LDL-R) activity. Different concentrations of ApoE2/3/4-DMPC complexes were added, respectively.ELISA was used to compare the ability of ApoE2/3/4 to competitively bind LDL-R, and RT-PCR was used to compare the effects of ApoE2/3/4 on LDLR mRNA and HMG-CoA mRNA expression after RSS administration.RESULTS In patients with ε2 (ε2/ε2, ε2/ε3), the levels of TC, TG, LDL-C, and Lp(a) decreased, and the level of HDL-C was significantly higher than that of ε3 group (ε3/ε3, ε2/ε4), and patients with ε4 have the worst efficacy; The relative expression of ApoE mRNA was significantly lower after treatment with RSS in three groups (P<0.05), but there was no statistically significant difference of ε3 (ε2/ε4) and ε4 (ε4/ε4) after treatment with RSS (P>0.05). In L-02 hepatocytes, ApoE2 binds to LDL-R more strongly than ApoE3 (P<0.05), and ApoE4 has the weakest binding to LDL-R. Compared with ApoE3, ApoE2 can up-regulate LDL-R mRNA expression(P<0.05), but ApoE4 inhibits LDLR mRNA expression (P<0.05); ApoE2 inhibited HMG-CoA mRNA expression significantly higher than ApoE3 (P<0.05), while ApoE4 inhibited HMG-CoA mRNA expression significantly lower than ApoE3 (P<0.05).CONCLUSION RSS has the best effect on lowering blood lipid in patients with ApoE ε2 allele, followed by ε3 and ε4 allele carriers, which may be related to the induction of LDLR gene expression by ApoE2, the enhancement of ApoE2 affinity to LDLR, and the enhancement of the inhibitory effect of RSS on HMG-CoA gene expression.