基因变异及心血管危险因素与单核细胞分泌白细胞介素6和10的关系

Relationship of genetic variants and cardiovascular risk factors with interleukin-6 and interleukin-10 secreted by monocytes

目的：研究白细胞介素（interleukin,IL）-6、IL-10基因变异及心血管危险因素[包括氧化低密度脂蛋白（oxygenized low density lipoprotein,ox-LDL）、缺乏体力活动、超重等]与单核细胞分泌IL-6和IL-10的关系.方法：2010年在北京社区人群随机抽取无心肌梗死和脑卒中病史的40名51 ～80岁的健康人,收集吸烟、饮酒资料,测量血压、空腹血糖、血脂等心血管危险因素指标,并检测IL-6（rs1800796、rs1524107、rs2066992）、IL-10基因变异（rs1800872、rs1554286、rs3021094）.分离静脉血单核细胞,在RPMI 1640培养液中培养24 h,分为二等份,一份加入ox-LDL（50 mg/L）刺激,另一份加入磷酸盐缓冲液（PBS）对照,继续培养48 h,采用ELISA法检测上清液IL-6和IL-10的浓度.结果：配对Wilcoxon检验显示,与PBS培养液相比,ox-LDL培养液中单核细胞分泌IL-6、IL-10以及IL-6/IL-10显著升高（P＜0.01）.以PBS和ox-LDL培养液中IL浓度作为重复测量结果,调整年龄、性别后,重复测量的多因素一般线性模型显示,超重者（BMI≥26 kg/m2）单核细胞分泌IL-10显著低于体重较轻者（P =0.007）,IL-6/IL-10则显著高于体重较低者（P=0.003）;缺乏体力活动者[metabolic equivalent of energy,METS＜ 166 kJ/（kg＆#183;d）]单核细胞分泌的IL-6/IL-10显著高于体力活动较高者（P=0.046）;饮酒者单核细胞分泌的IL-6和IL-10显著高于不饮酒者（P =0.049和0.006）;高密度脂蛋白胆固醇（high-density lipoprotein-eholesterol,HDL-c）较高者（≥56.4 mg/dL）单核细胞分泌IL-6显著升高（P=0.027）;IL-10的3个单核苷酸多态性（single nucleotide polymorphism,SNP）位点（rs1800872、rs1554286、rs3021094）与ox-LDL对单核细胞分泌IL-10存在显著交互作用（P＜0.05）,IL-6基因SNP与ox-LDL无显著交互作用.结论：ox-LDL、缺乏体力活动、超重等心血管因素与炎症-抗炎平衡向炎症方向偏斜密切相关,IL-10基因和ox-LDL的交互作用与抗炎细胞因子IL-10表达密切相关.

Objective：To examine the relationship of interleukin （IL）-6 and IL-10 genetic variants and cardiovascular factors [ oxygenized low density lipoprotein （ox-LDL）, lower physical activity, over- weight, etc. ] with IL-6 and IL-10 secreted by monocytes. Methods： In the study, 40 health persons, aged from 51 to 80 years, without stroke and myocardial infarction, were randomly sampled from a com- munity-based population in Beijing in 2010. Their data on smoking, drinking, blood pressure, fasting glucose, and lipid were collected. The single nucleotide polymorphisms （SNPs） of IL-6 （rs1800796, rs1524107, rs2066992） and IL-10 （rs1800872, rs1554286, rs3021094） were genotyped. The human monocytes were cultivated in RPMI 1640 medium for 24 h; then divided into two equal parts, in which ox-LDL （50 mg/L） and phosphate buffer solution （PBS） were added for another 48 h. Finally, the se- cretions of IL-6 and IL-10 in the culture supernatants were measured with ELISA. Results： Paired Wilcoxon tests showed that the IL-6, IL-10, and IL-6/IL-10 were significantly higher in ox-LDL medium than in PBS one （all P 〈 0.01 ）. The concentrations in PBS/ox-LDL taken as repeated measurements, and adjusted for age and gender, the repeated general linear models showed： IL-10 was significantly lower for those overweight （BMI〉~26 kg/m2） than for those normal weight （P = 0. 007 ）, and IL-6/IL- 10 was significantly higher in those overweight （P =0.003 ）. The IL-6/IL-10 was significantly higher in those with lower physical activity [ metabolic equivalent of energy, METS 〈 166 kJ/（ kg · d） ] than those with higher physical activities （P =0. 046）. IL-6 and IL-10 were significantly higher in alcohol drinkers （ P = 0. 049 and P = 0.006）. IL-6 was significantly higher in those with higher high-density lipoprotein- cholesterol （HDL-c, i〉 56.4 mg/dL, P = 0. 027 ）. There were significant interactions between IL-10 SNPs and ox-LDL on IL-10 （all P 〈 0.05 ）, but no significant interactions between IL-6 gene SNPs and ox-LDL on IL-6. Conclusion： The ox-LDL together with lower physical activity and overweight shifts the balance of pro-inflammatory and anti-inflammatory in the direction of pro-inflammatory. The interaction between IL-10 gene and ox-LDL is intensively correlated with the secretion of the anti-inflammatory cyto- kine IL-10.