中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2008年
3期
199-204
,共6页
田彩霞%李艳%汪明%夏尊恩%李晓艳%黄从新
田綵霞%李豔%汪明%夏尊恩%李曉豔%黃從新
전채하%리염%왕명%하존은%리효염%황종신
冠状动脉疾病%抗原,CD40%多态性,单核苷酸
冠狀動脈疾病%抗原,CD40%多態性,單覈苷痠
관상동맥질병%항원,CD40%다태성,단핵감산
Coronary disease%Antigens,CD40%Polymorphism,single nucleotide
目的 探讨急性冠状动脉综合征(ACS)与白细胞分化抗原40(CD40)、可溶性白细胞分化抗原40配体(sCD40L)、超敏C-反应蛋白(hs-CRP)的表达水平、白细胞计数、CD40-1C/T基因多态性的相关性,以及激活剂干扰素(IFN)-γ和抑制剂氟伐他汀对外周血单个核细胞CD40表达的影响.方法 随机选择160例经冠状动脉造影确诊的ACS患者和92例冠状动脉造影阴性人群,采用酶联免疫吸附试验、微粒子增强透射免疫分析技术、流式细胞分析技术检测血清sCD40L含量、血清hs-CRP,外周血白细胞计数水平.采用聚合酶链反应一限制性片段长度多态性(PCR-RFLP)和DNA测序技术,检测CD40-1C/T基因型和等位基因频率.随机选择40例冠状动脉造影结果为阳性的ACS患者,密度梯度离心法分离单个核细胞并原代培养,用100 ng/ml IFN-γ及10 μmol/L氟伐他汀对单个核细胞进行干预,流式细胞术检测单个核细胞培养24 h后表达CD40的水平.结果 ACS组的炎性细胞因子CD40[(4.59±2.07)MFI]、sCD40L[(7.58±2.34)μg/L]、hs-CRP[(5.24±1.34)mg/L、血白细胞计数水平(8.56×109/L±3.10×109/L)明显高于对照组[CD40:(2.08±1.19)MFI,sCD40L:(3.12±0.67)μg/L,hs-CRP:(0.88±0.90)mg/L,白细胞计数:6.12×109/L±1.41×109/L;均P＜0.01];ACS患者CC基因型频率为28.1%(45/160),明显高于对照组[13.0%(12/92),P＜0.01];C等位基因显著增加ACS的患病风险(OR=1.608,95%CI:1.12～2.32,P=0.011);基础状态下单个核细胞培养24 h后,CD40表达水平升高,且CC基因型单个核细胞CD40表达水平[(14.78±4.56)MFI]明显高于CT、TT基因型[(11.98±4.12)MFI,(9.86±3.83)MFI,P＜0.05];IFN-γ刺激状态下,单个核细胞表面CD40表达水平明显升高,且CC基因型升高的幅度(2.96倍)明显高于CT(2.37倍)和TT基因型(2.08倍),均P＜0.05;另外,氟伐他汀和IFN-γ共同作用,发现氟伐他汀可显著抑制IFN-γ诱导的CD40表达,但CC、CT、TT基因型单个核细胞经氟伐他汀干预后,其CD40表达水平差异无统计学意义(抑制率分别为30.3%、26.3%、29.3%,P＞0.05).结论 ACS的发生可激活炎症反应;CD40-1C/T基因多态性与ACS有相关性,C等位基因可能是ACS的易感相关基因;CD40-1C/T多态性可以影响单个核细胞CD40表达水平,氟伐他汀可显著抑制IFN-γ刺激后CD40的表达,但对不同基因型单个核细胞的保护作用无明显差异.
目的 探討急性冠狀動脈綜閤徵(ACS)與白細胞分化抗原40(CD40)、可溶性白細胞分化抗原40配體(sCD40L)、超敏C-反應蛋白(hs-CRP)的錶達水平、白細胞計數、CD40-1C/T基因多態性的相關性,以及激活劑榦擾素(IFN)-γ和抑製劑氟伐他汀對外週血單箇覈細胞CD40錶達的影響.方法 隨機選擇160例經冠狀動脈造影確診的ACS患者和92例冠狀動脈造影陰性人群,採用酶聯免疫吸附試驗、微粒子增彊透射免疫分析技術、流式細胞分析技術檢測血清sCD40L含量、血清hs-CRP,外週血白細胞計數水平.採用聚閤酶鏈反應一限製性片段長度多態性(PCR-RFLP)和DNA測序技術,檢測CD40-1C/T基因型和等位基因頻率.隨機選擇40例冠狀動脈造影結果為暘性的ACS患者,密度梯度離心法分離單箇覈細胞併原代培養,用100 ng/ml IFN-γ及10 μmol/L氟伐他汀對單箇覈細胞進行榦預,流式細胞術檢測單箇覈細胞培養24 h後錶達CD40的水平.結果 ACS組的炎性細胞因子CD40[(4.59±2.07)MFI]、sCD40L[(7.58±2.34)μg/L]、hs-CRP[(5.24±1.34)mg/L、血白細胞計數水平(8.56×109/L±3.10×109/L)明顯高于對照組[CD40:(2.08±1.19)MFI,sCD40L:(3.12±0.67)μg/L,hs-CRP:(0.88±0.90)mg/L,白細胞計數:6.12×109/L±1.41×109/L;均P＜0.01];ACS患者CC基因型頻率為28.1%(45/160),明顯高于對照組[13.0%(12/92),P＜0.01];C等位基因顯著增加ACS的患病風險(OR=1.608,95%CI:1.12～2.32,P=0.011);基礎狀態下單箇覈細胞培養24 h後,CD40錶達水平升高,且CC基因型單箇覈細胞CD40錶達水平[(14.78±4.56)MFI]明顯高于CT、TT基因型[(11.98±4.12)MFI,(9.86±3.83)MFI,P＜0.05];IFN-γ刺激狀態下,單箇覈細胞錶麵CD40錶達水平明顯升高,且CC基因型升高的幅度(2.96倍)明顯高于CT(2.37倍)和TT基因型(2.08倍),均P＜0.05;另外,氟伐他汀和IFN-γ共同作用,髮現氟伐他汀可顯著抑製IFN-γ誘導的CD40錶達,但CC、CT、TT基因型單箇覈細胞經氟伐他汀榦預後,其CD40錶達水平差異無統計學意義(抑製率分彆為30.3%、26.3%、29.3%,P＞0.05).結論 ACS的髮生可激活炎癥反應;CD40-1C/T基因多態性與ACS有相關性,C等位基因可能是ACS的易感相關基因;CD40-1C/T多態性可以影響單箇覈細胞CD40錶達水平,氟伐他汀可顯著抑製IFN-γ刺激後CD40的錶達,但對不同基因型單箇覈細胞的保護作用無明顯差異.
목적 탐토급성관상동맥종합정(ACS)여백세포분화항원40(CD40)、가용성백세포분화항원40배체(sCD40L)、초민C-반응단백(hs-CRP)적표체수평、백세포계수、CD40-1C/T기인다태성적상관성,이급격활제간우소(IFN)-γ화억제제불벌타정대외주혈단개핵세포CD40표체적영향.방법 수궤선택160례경관상동맥조영학진적ACS환자화92례관상동맥조영음성인군,채용매련면역흡부시험、미입자증강투사면역분석기술、류식세포분석기술검측혈청sCD40L함량、혈청hs-CRP,외주혈백세포계수수평.채용취합매련반응일한제성편단장도다태성(PCR-RFLP)화DNA측서기술,검측CD40-1C/T기인형화등위기인빈솔.수궤선택40례관상동맥조영결과위양성적ACS환자,밀도제도리심법분리단개핵세포병원대배양,용100 ng/ml IFN-γ급10 μmol/L불벌타정대단개핵세포진행간예,류식세포술검측단개핵세포배양24 h후표체CD40적수평.결과 ACS조적염성세포인자CD40[(4.59±2.07)MFI]、sCD40L[(7.58±2.34)μg/L]、hs-CRP[(5.24±1.34)mg/L、혈백세포계수수평(8.56×109/L±3.10×109/L)명현고우대조조[CD40:(2.08±1.19)MFI,sCD40L:(3.12±0.67)μg/L,hs-CRP:(0.88±0.90)mg/L,백세포계수:6.12×109/L±1.41×109/L;균P＜0.01];ACS환자CC기인형빈솔위28.1%(45/160),명현고우대조조[13.0%(12/92),P＜0.01];C등위기인현저증가ACS적환병풍험(OR=1.608,95%CI:1.12～2.32,P=0.011);기출상태하단개핵세포배양24 h후,CD40표체수평승고,차CC기인형단개핵세포CD40표체수평[(14.78±4.56)MFI]명현고우CT、TT기인형[(11.98±4.12)MFI,(9.86±3.83)MFI,P＜0.05];IFN-γ자격상태하,단개핵세포표면CD40표체수평명현승고,차CC기인형승고적폭도(2.96배)명현고우CT(2.37배)화TT기인형(2.08배),균P＜0.05;령외,불벌타정화IFN-γ공동작용,발현불벌타정가현저억제IFN-γ유도적CD40표체,단CC、CT、TT기인형단개핵세포경불벌타정간예후,기CD40표체수평차이무통계학의의(억제솔분별위30.3%、26.3%、29.3%,P＞0.05).결론 ACS적발생가격활염증반응;CD40-1C/T기인다태성여ACS유상관성,C등위기인가능시ACS적역감상관기인;CD40-1C/T다태성가이영향단개핵세포CD40표체수평,불벌타정가현저억제IFN-γ자격후CD40적표체,단대불동기인형단개핵세포적보호작용무명현차이.
Objective To investigate the expression levels of CD40,sCD40L,hs-CRP,WBC in acute coronary syndrome ( ACS)patients and the association between CD40-1C/T single nucleotide polymorphism and risk of ACS in Han Chinese,moreover,the regulatory effects of IFN-γ and fluvastatin on the expression of CD40 in peripheral blood mononuclear cell(PBMNC)were also observed.Methods (1) 160 ACS patients and 92 control patients diagnosed by coronary angiography were recruited.Enzyme linked immunosorbent assay,particle enhanced immunoturbidimetric assay,flow cytometry were used to detect the levels of soluble CD40L,hs-CRP,and WBC count.(2)The CD40 genotype and allele frequency were analyzed by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)and DNA sequencing technology.(3)PBMNC were separated by density gradient centrifugating heparinized venous blood from 40 ACS patients,cultured for 24 h with or without 100 ng/ml IFN-γ in the absence or present of 10 μmol/L fluvastatin.The CD40 expression levels were then detected by flow cytometry. Results Inflammatory cytokine CD40、sCD40L、hs-CRP levels were significantly higher in ACS patients than in controls.The CD40-1C allele frequency was 0.606 in ACS group and 0.489 in controls,while T allele frequency was 0.394 in ACS group and 0.511 in controls.The frequency of CC genotype was significantly higher in ACS group than in controls(P＜0.01).C allele carriers had significantly higher risk of ACS (OR=1.608,95%CI:1.12-2.32,P=0.011).CD40 production increased after 24 h culturing and the CD40 levels were significantly higher in subiects with CC genotype than that in subiects with CT or TF genotype[CC:(14.78 ±4.56)MFI,CT:(11.98±4.12)MFI,TT:(9.86±3.83)MFI,P＜0.05],IFN-γ further increased PBMNC CD40 expressions in all subjects after culturing for 24 h and fluvastatin equally inhibited IFN-γ induced PBMNC CD40 expression from various genotypes(CC,CT,TT was 30.3%,26.3%,29.3% respectively,all P＞0.05).Conclusion Inflammatory cytokines were increased in ACS patients and CD40-1C/T polymorphism iS associated with higher risk for ACS in Han Chinese.
