摘要目的 探讨骨保护素(OPG)启动子区域T149C、T950C基因多态性及血清OPG、可溶性核转录因子-κB受体活化因子配体(sRANKL)含量与冠心病(CHD)发病风险的关系.方法 选择2017年4月至2018年12月天津市胸科医院心内科收治的天津地区疑似CHD入院并接受冠状动脉造影(CAG)的528例患者,根据CAG检查结果将患者分为CHD组(302例)和非CHD组(226例).记录所有受试对象的性别、年龄、高血压史、CHD家族史、糖尿病、血脂指标等临床资料;采用酶联免疫吸附试验(ELISA)检测血清OPG、sRANKL水平;应用聚合酶链反应-限制性内切酶片段长度多态性(PCR-RFLP)技术检测T149C、T950C基因多态性,等位基因进行Hardy-Weinberg遗传平衡检验;采用二分类多因素非条件Logistic回归法分析T149C、T950C基因多态性及血清OPG、sRANKL水平与CHD的关系.结果 所有患者均纳入最终分析.CHD组血清OPG水平明显高于非CHD组(μg/L :1.76±0.49比1.47±0.29,P＜0.01),血清sRANKL水平明显低于非CHD组(ng/L :342.14±121.38比376.63±108.66,P＜0.05).Logistic回归分析显示,在调整了年龄、性别、血脂指标、糖尿病等因素后,血清OPG升高为发生CHD的独立危险因素〔优势比(OR)＝1.995,95%可信区间(95%CI)＝1.935～2.066,P＝0.012〕.PCR-RFLP检测结果显示,OPG基因启动子区域T149C、T950C均有TT、TC、CC 3种基因型.经Hardy-Weinberg平衡检验,OPG T149C及OPG T950C基因多态性符合Hardy-Weinberg定律,达到遗传平衡,具有群体代表性.非CHD组T149C基因型TT、TC、CC及等位基因T、C的频率分别为53.5%、42.9%、3.6%、75.0%和25.0%,CHD组分别为43.1%、50.3%、6.6%、68.2%和31.8%,两组间基因型和等位基因频率比较差异均有统计学意义(均P＜0.05).Logistic回归分析显示,T149C位点的TC+CC基因型发生CHD的风险是TT基因型的1.86倍(OR＝1.86,95%CI＝1.24～2.78,P＝0.003),提示C等位基因可能为发生CHD的易感基因.非CHD组T950C基因型TT、TC、CC及等位基因T、C的频率分别为39.8%、46.5%、13.7%、63.1%和36.9%,CHD组分别为39.4%、43.4%、17.2%、61.1%和38.9%,两组间基因型和等位基因频率比较差异均无统计学意义(均P＞0.05).Logistic回归分析显示,T950C位点的TC+CC基因型与CHD的发生无关.结论 血清OPG水平升高与CHD发生有关,可作为CHD发生的危险因素.OPG T149C的TC+CC基因型与CHD发生密切相关,C等位基因可能为发生CHD的易感基因.

abstractsObjective To explore the polymorphisms of T149C and T950C gene in osteoprotectin (OPG) promoter sites and the levels of serum OPG and soluble nuclear factor-κB receptor activator ligand (sRANKL) and the incidence of coronary heart disease (CHD). Methods 528 patients in Tianjin suspected of CHD and underwent coronary angiography (CAG) who admitted to the department of cardiology of Tianjin Chest Hospital from April 2017 to December 2018 were enrolled. According to the CAG results, they were divided into two groups: CHD group (n = 302) and non-CHD group (n = 226). The gender, age, history of hypertension, family history of CHD, diabetes, levels of blood lipid parameters in serum and other clinical data of patients were recorded. The levels of serum OPG and sRANKL were measured by enzyme-linked immunosorbent assay (ELISA). T149C and T950C gene polymorphisms were analyzed by polymerase chain reaction-restriction endonuclease fragment length polymorphism (PCR-RFLP) methods. Hardy-Weinberg genetic balance test was performed for alleles. Binomial classification multivariate non-conditional Logistic regression method was used to analyze the relationship between T149C and T950C gene polymorphisms, serum levels of OPG and sRANKL and CHD. Results All patients were enrolled in the final analysis. The serum level of OPG in CHD group was significantly higher than that in non-CHD group (μg/L: 1.76±0.49 vs. 1.47±0.29, P < 0.01), the serum level of sRANKL was significantly lower than that in non-CHD group (ng/L: 342.14±121.38 vs. 376.63±108.66, P < 0.05). Logistic regression analysis showed that after adjusting for age, gender, blood lipid parameters, diabetes and other factors, the increase in serum OPG level was an independent risk factor for CHD [odds ratio (OR) = 1.995, 95% confidence interval (95%CI) = 1.935-2.066, P = 0.012]. PCR-RFLP results showed that TT, TC and CC genotypes were found in T149C and T950C of OPG promoter. According to Hardy-Weinberg equilibrium test, the polymorphisms of OPG T149C and T950C accorded with Hardy-Weinberg law, achieving genetic balance with representative of the population. The frequencies of TT, TC, CC and alleles T and C in T149C genotypes of non-CHD group were 53.5%, 42.9%, 3.6%, 75.0% and 25.0%, respectively, and they were 43.1%, 50.3%, 6.6%, 68.2% and 31.8%, respectively in CHD group. There were statistically significant differences in genotype and allele frequencies between the two groups (all P < 0.05). It was shown by Logistic regression analysis that the risk of CHD in TC+CC genotype of T149C was 1.86 of TT genotype (OR = 1.86, 95%CI = 1.24-2.78, P = 0.003). It was suggested that C allele might be a susceptible gene for CHD. In non-CHD group, the frequencies of TT, TC, CC, and alleles T and C in T950C genotypes were 39.8%, 46.5%, 13.7%, 63.1% and 36.9%, respectively. They were 39.4%, 43.4%, 17.2%, 61.1% and 38.9%, respectively in CHD group. There were no significant differences in genotype and allele frequencies between the two groups (all P > 0.05). Logistic regression analysis showed that TC+CC genotype of T950C was not related with CHD. Conclusions The increased level of serum OPG was closely related with CHD and could be used as a risk factor for CHD. The cases carried OPG T149C TC+CC genotype might have the risk suffering CHD. C allele is might be a susceptible gene.