低密度脂蛋白胆固醇抗体对急性冠脉综合征患者的保护机制

Protection mechanism of autoantibodies against ox-LDL in ACS patients

目的阐明急性冠脉综合征(ACS)患者急性期趋化因子配体[chemokine(C-C motif)ligand,CCL)5和白细胞介素(IL)-18波动与ox-LDL自身抗体的关系,T淋巴细胞的免疫漂移规律;以及观察不同的再灌注治疗策略对减轻炎症反应的作用。方法入选对照组(n=30):冠状动脉造影阴性人员。随机序贯入选60位ACS患者,将其分为两组:早期再灌注(ER,n=29)组和晚期再灌注(LR,n=31)组。所有患者均进行冠脉造影,并采集入院后第2天至第28天的静脉血液样本,测定指标包括:血脂水平:总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)和ox-LDL,炎症因子:CCL5、IL-18、IL-33、IL-5和超敏C反应蛋白(hs-CRP),血浆免疫球蛋白(Ig)亚型,及分析Th1[CD4+γ干扰素(IFN-γ)+]和Th2(CD4+IL-4+)细胞亚群。结果 (1)炎症因子:CCL5:ER组在第8天出现下降,而LR组在第14天出现下降,IL-18:ER组在第8天达到峰值,而LR组则在第12天达到峰值,IL-5和IL-33:ER组的曲线拐点出现在第4天,而LR组出现在第16天,hs-CRP:ER组的第4天出现下降而LR组在第14天出现下降。(2)血脂水平:4周内对照组的血脂水平(TC、TG、LDL和oxLDL)没有明显变化,两种不同的再灌注策略中,血脂没有明显差异。(3)免疫漂移:ER组和LR组第1周T淋巴细胞免疫漂移最为显著。Th1漂移出现明显缓解,Th1漂移现象分别在ER组第2周和LR组第3周开始出现,并于第4周两组的Th1/Th2值最终趋于相等。与LR组比较,ER组前2周产生更多总Ig A、Ig E、Ig G1,但两组在总Ig G2a和Ig M水平上几乎完全相等。重要的是,前两周中ER组中ox-LDL的Ig G1抗体显著升高,而在ox-LDL的Ig M抗体方面,ER和LR组之间没有显著差异。(4)相关性和线性回归分析:上述连续变量的相关性研究发现,血浆浓度CCL5与IL-18,IL-33与IL-5明显正相关性(P<0.01)。多元线性回归分析表明,CCL5、ox-LDL和hs-CRP正相关,而IL-33、ox-LDL抗体与hs-CRP水平负相关。结论与LR治疗相比,ER治疗策略能明显降低不稳定斑块的附加风险,可能优于晚期再灌注治疗。

AIM The first aim was to clarify whether CCL5 and IL-18 are accompanied with autoantibodies against ox-LDL activity and T-lymphocyte immune drift in ACS patients. A secondary aim was to investigate the extent of different reperfusion therapeutic strategies to alleviate inflammation. METHODS The present investigation recruited 30 coronary angiography negative persons as ①control group （n = 30） , and studied a total of 60 consecutive unselected ACS patients divided into other two groups ： ②early reperfusion （ER） group and ③Late reperfusion （LR, after 2 weeks） combinated group. All patients had coronary angiography and venous blood samples of all participants were drawn every morning from day 2 to 28 after admission. Detection index including： blood fat index （TC, TG, HDL-C, LDL-C and ox-LDL）, inflam- mation index （CCLS, IL-18, IL-33, IL-5 and hs-CRP） , Plasma Ig isotypes, and analysis of Thl （CIM ＋ IFNγ＋ ）, Th2 （ CD4 ＋ IL4 ＋ ） by flow cytometry. RESULTS ①Blood lipid fluctuation ： The blood lipid levels （TC, TG, LDL and ox-LDL） of the control group had little change during the 4 weeks （P 〉 0. 05 ）. Between the two different reperfusion strategies, there was no differences in improvement of blood lipids metabolism （P 〉 0. 05 ）. ②Inflammatory cytokines dynamic state; CCL5 decreases in the ER group were demonstrated at day 8, but in the LR group not until day 14. IL-18 reached a maximum at day 8 in the ER group but peaked at day 12 in the LR group. IL-5 and IL-33 appeared at day 4 in the ER group but were seen at day 16 in the LR group, hsCRP decreased in the ER group at day 4 but not until day 14 in the LR group. ③T-lymphocyte immune drift state： Flow cytometry showed that T lymphocyte subsets： both in the ER group and the LR group, T-lymphocyte immune drift was the most significant in the first week. Thl bias who was relieved markedly from the beginning of the second week in ER group and from the third week in LR group evolved eventually toward an almost equal value of Th1/Th2 in the two treatment groups at the fourth week. The results showed that the ER group produced more total IgA, IgE, and IgG1 in the first 2 weeks, but total IgG2a and IgM in the ER group were almost equal with that in the LR group. Importantly, significantly elevated IgG1 antibody of ox-LDL were demonstrated at the ER group in the first 2 weeks, but IgM antibody of ox-LDL did not demonstrate significant differences between the ER and the LR groups. ④Correlation analysis was performed for continuous variables and found that plasma concentration of CCI5 with IL-18, IL-33 with IL-5 had clear association （P 〈0. 01 ）. Multiple linear regression analysis showed that CCL5 and ox-LDL had positive relationships with hsCRP, but IL-33 and ox-LDL antibody maintained negtive predictors with hs-CRP. CONCLUSION EarIy reperfusion strategy significantly alleviates the additional risk of plaque vulnerability compared with late reperfusion treatment and efficacy of early reperfusion strategy maybe better than late reperfusion treatment.