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随着生活方式及生活习惯的改变,心血管疾病发病率不断升高,已经成为全球过早死亡的主要原因[1-2]。冠心病(coronary heart disease,CHD)是冠状动脉粥样硬化性心脏病的简称,它是心血管系统中最常见、最主要的疾病,是由冠状动脉(冠脉)狭窄或者供血不足而引起的心肌功能障碍的一种器质性病变,是全世界致死和致残的主要原因。
乙醛脱氢酶2(acetaldehyde dehydrogenase 2, ALDH2)作为一种线粒体特异性酶,可特异性地将体内细胞毒性的乙醛转化成无毒的乙酸,减少生物膜中磷脂双分子层中脂质过度氧化,减轻膜蛋白的变形、线粒体DNA的损伤及细胞内氧化应激[3-4]。近年来相关研究[5-7]发现ALDH2基因多态性与CHD密切相关。NLRP3炎症体是炎症机制和细胞程序性死亡的重要的信号通路。相关研究[8]显示,当其被特定的内外源性信号激活后可诱发炎症体活化从而引起细胞焦亡及炎性产物(IL-1β、IL-18)等物质产生,并可在CHD病程进展中发挥作用[8]。近期研究[9]发现,ALDH2可促进体内细胞氧化应激及炎症介质(ROS等)产生从而促进炎症小体NLRP3的表达,该研究为我们提供线索,即ALDH2是否通过NLRP3调控炎症反应从而在CHD中发挥作用。
现阶段,ALDH2及NLRP3在CHD中的具体治病机制尚不明确,血浆中ALDH2及NLRP3水平与CHD的相关性仍未被充分研究,本研究拟通过检测不同病情CHD病人血浆中ALDH2及NLRP3活性水平,明确ALDH2及NLRP3活性水平与CHD病人病情变化之间的关系,为CHD在临床上的诊断治疗提供可供参考的依据。
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收集2018年1月至2019年6月蚌埠医学院第一附属医院诊断明确患有CHD的病人共170例,男81例,女89例,年龄(63.84±9.0)岁。其诊断分型依照1999年WHO/WPR标准,所有病人均接受常规12导联心电图或动态心电图、心脏彩超和冠脉造影等检查明确病情。所有病人入院后完善血常规、生化常规检查并记录血压和体质量指数。CHD病人按照冠脉造影中记录的冠脉左主干(LM)、左前降支(LAD)、左旋支(LCX)、右冠支(RCA)病变情况,依据狭窄程度最严重的病变血管情况分为:轻度狭窄(is-CHD,狭窄程度 < 50%)组19例、中度狭窄(ms-CHD,狭窄程度50%~75%)组41例和严重狭窄(ss-CHD,狭窄程度≥75%)组73例[10]。其中有胸痛发作症状但完善相关检查并行冠脉血管造影未见明显异常者作为对照组共37例。排除标准:既往有心肌炎、心肌病、结构性心脏病、恶性肿瘤类疾病、风湿免疫系统疾病、内分泌及代谢性疾病,或近期服用降低自身免疫功能的药物,神经系统退行性改变疾病、脑卒中、心房颤动及感染性疾病[11]。Gensini评分法[12]见表 1。
狭窄程度/% 评分/分 病变部分 评分/分 1~<25 1 左主干 5 25~ < 50 2 左前降支、左回旋支近段 2.5 50~ < 75 4 左前降支中段 1.5 75~ < 90 8 左前降支远段、第一对角支 1 90~ < 100 16 左回旋支远段、后降支 1 100 32 右冠脉 1 小分支 0.5 表 1 冠脉Gensini评分
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记录研究对象的姓名、性别、年龄、有无饮酒习惯、吸烟史等一般信息,有无高血压、糖尿病等既往疾病史。
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入选对象禁食12h后抽取肘静脉血3mL于肝素抗凝试管中,3000 r/min离心5 min,吸取上清,-80℃冰箱中备用。
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标本彻底融化后,采用ELISA法,按上海杰美基因医药科技有限公司的试剂盒说明书进行操作,分别取样本检测ALDH2及NLRP3 OD值,代入标准品与OD值之间的回归曲线方程,计算浓度值。
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采用方差分析、q检验和Pearson相关分析。
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ss-CHD组血肌酐(CR)水平高于对照组和ms-CHD组(P < 0.05),高密度脂蛋白(HDL)在ss-CHD组中明显低于对照组(P < 0.01),ms-CHD和ss-CHD组淋巴/单核细胞低于is-CHD组(P < 0.05)。而各组间三酰甘油(TG)、胆固醇(TC)、总蛋白及低密度脂蛋白(LDL)、红细胞(WBC)、淋巴/单核细胞差异无统计学意义(P>0.05)(见表 2)。
分组 n 年龄/岁 TC/(mmol/L) TG/(mmol/L) HDL/(mmol/L) LDL/(mmol/L) 总蛋白/(g/L) ALDH2/(U/L) NLRP3/(ng/mL) CR/(μmol/L) WBC/(×109) 淋巴/单核细胞/(×109) 对照组 37 57.81±10.49 4.08±1.23 1.76±1.31 1.19±1.07 2.22±0.70 67.28±6.01 138.30±36.36 4.15±1.19 61.62±11.50 6.26±2.09 5.13±2.40 is-CHD组 19 61.68±11.78 4.39±1.16 1.77±1.07 1.04±0.20 2.42±0.78 66.71±6.21 131.61±29.66 5.30±1.75 66.37±9.85 6.12±1.08 6.61±7.14 ms-CHD组 41 64.76±10.22 4.25±1.12 1.44±0.65 1.09±0.25 2.40±0.90 66.21±6.44 117.76±30.24* 5.49±2.38** 63.68±12.13 6.41±1.86 4.73±1.80# ss-CHD组 73 64.85±9.61** 4.19±1.74 1.81±2.01 0.90±0.23** 2.25±0.73 65.52±0.76 109.57±29.82**## 7.28±2.69**##△△ 73.08±27.15**△△ 6.84±2.02 4.74±2.35# F — 4.50 0.21 0.55 2.81 0.60 0.80 17.26 7.79 3.53 1.25 1.93 P — < 0.01 >0.05 >0.05 < 0.05 >0.05 >0.05 < 0.01 < 0.01 < 0.05 >0.05 >0.05 MS组内 — 104.149 2.098 2.357 0.293 0.595 40.410 5.137 987.940 394.383 3.669 9.962 q检验:与对照组比较*P < 0.05,**P < 0.01;与is-CHD组比较#P < 0.05,##P < 0.01;与ms-CHD组比较△P < 0.05,△△P < 0.01 表 2 不同严重程度CHD病人临床指标比较(x±s)
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Is-CHD组、ms-CHD组和ss-CHD组血清ALDH2水平均较对照组减低(P < 0.05~ P < 0.01),并且随着冠脉狭窄严重程度的加重而逐渐减低(P < 0.01);而血清NLRP3水平随着冠脉病变程度加重其含量逐渐升高(P < 0.01),且两者之间呈正相关关系(P < 0.05)(见表 2)。
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根据病人冠脉造影结果将CHD病人分为单支、双支及多支病变(三支及以上),结果显示:随着CHD病变支数的增加ALDH2逐渐下降(P < 0.05~P < 0.01),而NLRP3水平逐渐增加(P < 0.05~P < 0.01),以NLRP3变化为显著(见表 3)。
分组 ALDH2/(U/L) NLRP3/(ng/mL) 对照组 136.70±36.40 4.21±1.25 单支病变组 121.32±28.20* 5.47±2.44* 双支病变组 110.05±32.63** 6.65±2.65**# 多支病变组 117.81±29.56* 7.18±2.59**## F 5.45 12.53 P < 0.01 < 0.01 MS组内 936.503 6.630 q检验:与对照组比较*P < 0.05,**P < 0.01;与单支病变比较#P < 0.05,##P < 0.01;与双支病变比较△P < 0.05,△△P < 0.01 表 3 血清ALDH2及NLRP3活性水平与病变支数的关系(x±s)
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血清ALDH2水平与Gensini评分呈明显负相关关系(r=-0.247,P < 0.01),而NLRP3水平与Gensini评分呈明显正相关关系(r=0.477,P < 0.01)。血清ALDH2与LDL呈负相关关系(r=-0.220,P < 0.05);而血清NLRP3与ALDH2呈负相关关系(r=-0.201,P < 0.05)。
血清ALDH2和NLRP3活性水平在冠心病不同血管病变程度中的变化
Changes of the serum ALDH2 and NLRP3 activity levels in different degrees of coronary artery disease
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摘要:
目的通过检测血清线粒体乙醛脱氢酶2(ALDH2)及核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)活性水平,分析血清ALDH2和NLRP3与冠心病(CHD)不同病变程度之间的相关性,探讨二者在CHD发生、发展中的变化规律。 方法选择因胸痛入院并行冠脉造影检查的病人170例,其中冠状动脉(冠脉)造影明确诊断为CHD病人133例,同期冠脉造影正常者37例(对照组)。133例CHD病人按照冠脉狭窄严重程度分成轻度狭窄组(< 50%,is-CHD组)19例、中度狭窄组(50%~75%,ms-CHD组)41例和严重狭窄组(≥75%,ss-CHD组)73例。应用酶联免疫吸附实验检测血清中ALDH2及NLRP3活性水平,分析在有无CHD及冠脉不同病变程度下ALDH2和NLRP3活性差异,探讨ALDH2及NLRP3活性水平与CHD发生、发展的关系,采用Pearson相关性分析ALDH2和NLRP3的血清学影响因素。 结果CHD组血肌酐水平高于对照组和ms-CHD组(P < 0.05),高密度脂蛋白在ss-CHD组中明显低于对照组(P < 0.01),ms-CHD和ss-CHD组淋巴/单核细胞低于is-CHD组(P < 0.05)。is-CHD组、ms-CHD组和ss-CHD组血清ALDH2水平均较对照组减低(P < 0.05~P < 0.01),并且随着冠脉狭窄严重程度的加重而逐渐减低(P < 0.01);而血清NLRP3水平随着冠脉病变程度加重其含量逐渐升高(P < 0.01),且两者之间呈正相关关系(P < 0.05)。随着CHD病变支数的增加ALDH2逐渐下降(P < 0.01),而NLRP3水平逐渐增加(P < 0.05),以NLRP3变化为显著。血清ALDH2水平与Gensini评分呈明显负相关关系(r=-0.247,P < 0.01),而NLRP3水平与Gensini评分呈明显正相关关系(r=0.477,P < 0.01)。血清ALDH2与LDL呈负相关关系(r=-0.220,P < 0.05);而血清NLRP3与ALDH2呈负相关关系(r=-0.201,P < 0.05)。 结论随着冠脉病变程度加重,血清ALDH2水平逐渐降低,血清NLRP3水平逐渐升高,两者间存在一定的线性相关。 Abstract:ObjectiveTo detect the activity levels of serum mitochondrial acetaldehyde dehydrogenase 2 (ALDH2) and nucleotide-binding oligomerization domain-like receptors protein 3 (NLRP3), analyze the correlations between ALDH2, NLRP3 and different courses of coronary heart disease (CHD), and explore the change rule of both in the occurrence and development of CHD. MethodsA total of 170 chest pain patients detected by coronary angiography were investigated, and 133 CHD patients and 37 normal patients (control group) were identified.One hundred and thirty-three CHD patients were divided into the mild stenosis group (19 cases with stenosis deree of < 50%, is-CHD group), moderate stenosis group (41 cases degree of stenosis with 50% to 75%, ms-CHD group) and severe stenosis group (73 cases with stenosis degree of ≥75%, ss-CHD group) according to the severity of coronary artery stenosis.The activity levels of ALDH2 and NLRP3 in all cases were detected using enzyme-linked immunosorbent assay.The differences of the activity levels of ALDH2 and NLRP3 in CHD patients with different degrees of disease were analyzed, the correlations between activity levels of ALDH2, NLRP3 and occurrence, development of CHD were disscussed.The serological influencing factors of ALDH2 and NLRP3 were analyzed using the Pearson correlation analysis. ResultsThe serum level of creatinine in CHD group was higher than that in control group and ms-CHD group (P < 0.05), the high-density lipoprotein level in ss-CHD group was significantly lower than that in control group (P < 0.01), while the lymphocyte/monocytes in ms-CHD group and ss-CHD group were lower than that in the is-CHD group (P < 0.05).The serum levels of ALDH2 in ms-CHD and ss-CHD groups were lower than that in control group, which gradually decreased with the aggravation of the severity of coronary artery stenosis (P < 0.05-P < 0.01), while the serum level of NLRP3 gradually increased with the aggravation of the degree of coronary artery disease (P < 0.01), and there was a positive correlation between both (P < 0.05).With the increasing of the number of CHD lesions, the level of ALDH2 gradually decreased (P < 0.01), while the level of NLRP3 gradually increased (P < 0.05), and the change of NLRP3 was significant.The serum level of ALDH2 was negatively correlated with Gensini score (r=-0.247, P < 0.01), while the NLRP3 level was positively correlated with Gensini score (r=0.477, P < 0.01).The serum level of ALDH2 was negatively correlated with LDL (r=-0.220, P < 0.05) and the serum level of NLRP3 was negatively correlated with ALDH2 (r=-0.201, P < 0.05). ConclusionWith the aggravation of coronary artery disease, the serum level of ALDH2 gradually decreases, the serum level of NLRP3 gradually increases, and there was a certain correlation between them. -
表 1 冠脉Gensini评分
狭窄程度/% 评分/分 病变部分 评分/分 1~<25 1 左主干 5 25~ < 50 2 左前降支、左回旋支近段 2.5 50~ < 75 4 左前降支中段 1.5 75~ < 90 8 左前降支远段、第一对角支 1 90~ < 100 16 左回旋支远段、后降支 1 100 32 右冠脉 1 小分支 0.5 表 2 不同严重程度CHD病人临床指标比较(x±s)
分组 n 年龄/岁 TC/(mmol/L) TG/(mmol/L) HDL/(mmol/L) LDL/(mmol/L) 总蛋白/(g/L) ALDH2/(U/L) NLRP3/(ng/mL) CR/(μmol/L) WBC/(×109) 淋巴/单核细胞/(×109) 对照组 37 57.81±10.49 4.08±1.23 1.76±1.31 1.19±1.07 2.22±0.70 67.28±6.01 138.30±36.36 4.15±1.19 61.62±11.50 6.26±2.09 5.13±2.40 is-CHD组 19 61.68±11.78 4.39±1.16 1.77±1.07 1.04±0.20 2.42±0.78 66.71±6.21 131.61±29.66 5.30±1.75 66.37±9.85 6.12±1.08 6.61±7.14 ms-CHD组 41 64.76±10.22 4.25±1.12 1.44±0.65 1.09±0.25 2.40±0.90 66.21±6.44 117.76±30.24* 5.49±2.38** 63.68±12.13 6.41±1.86 4.73±1.80# ss-CHD组 73 64.85±9.61** 4.19±1.74 1.81±2.01 0.90±0.23** 2.25±0.73 65.52±0.76 109.57±29.82**## 7.28±2.69**##△△ 73.08±27.15**△△ 6.84±2.02 4.74±2.35# F — 4.50 0.21 0.55 2.81 0.60 0.80 17.26 7.79 3.53 1.25 1.93 P — < 0.01 >0.05 >0.05 < 0.05 >0.05 >0.05 < 0.01 < 0.01 < 0.05 >0.05 >0.05 MS组内 — 104.149 2.098 2.357 0.293 0.595 40.410 5.137 987.940 394.383 3.669 9.962 q检验:与对照组比较*P < 0.05,**P < 0.01;与is-CHD组比较#P < 0.05,##P < 0.01;与ms-CHD组比较△P < 0.05,△△P < 0.01 表 3 血清ALDH2及NLRP3活性水平与病变支数的关系(x±s)
分组 ALDH2/(U/L) NLRP3/(ng/mL) 对照组 136.70±36.40 4.21±1.25 单支病变组 121.32±28.20* 5.47±2.44* 双支病变组 110.05±32.63** 6.65±2.65**# 多支病变组 117.81±29.56* 7.18±2.59**## F 5.45 12.53 P < 0.01 < 0.01 MS组内 936.503 6.630 q检验:与对照组比较*P < 0.05,**P < 0.01;与单支病变比较#P < 0.05,##P < 0.01;与双支病变比较△P < 0.05,△△P < 0.01 -
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