中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
12期
895-898
,共4页
庞琪%刘晓菊%施凯%曾晓丽%包海荣
龐琪%劉曉菊%施凱%曾曉麗%包海榮
방기%류효국%시개%증효려%포해영
肺疾病,慢性阻塞性%巨噬细胞%香烟烟雾提取物%吞噬%氧化应激
肺疾病,慢性阻塞性%巨噬細胞%香煙煙霧提取物%吞噬%氧化應激
폐질병,만성조새성%거서세포%향연연무제취물%탄서%양화응격
Pulmonary disease,chronic obstructive%Macrophages%Cigarette smoke extract%Phagocytosis%Oxidative stress
目的 探讨香烟烟雾提取物(CSE)对慢性阻塞性肺疾病(简称慢阻肺)患者单核细胞源性巨噬细胞(MDM)吞噬功能的影响.方法 选择2012年1月至2013年3月兰州大学第一医院慢阻肺稳定期患者32例,选择同期健康体检者32名,分离外周血单核细胞,体外诱导培养成MDM.分别将慢阻肺患者和健康体检者的MDM分为慢阻肺非CSE组(常规培养)、慢阻肺CSE组(4％ CSE干预6h)、健康非CSE组(常规培养)、健康CSE组(4％ CSE干预6h).以流式细胞术的平均荧光强度和激光共聚焦显微镜的荧光灰度值判断MDM吞噬荧光标记大肠杆菌(FITC-E.coli的能力,菲罗啉比色法检测细胞上清液总抗氧化能力(TAC),硫代巴比妥酸比色法检测丙二醛含量,改良Hafeman直接测定法检测谷胱甘肽过氧化物酶(GSH-PX)活力.结果 平均荧光强度、荧光灰度值:慢阻肺非CSE组(20.2±2.2、51.5±5.8)显著低于健康非CSE组(56.9±6.7、87.3 ±7.3),慢阻肺CSE组(7.6±0.7、14.1 ±3.4)和健康CSE组(48.0±5.4、69.7±6.0)分别显著低于慢阻肺非CSE组和健康非CSE组(均P＜0.01).TAC、GSH-PX水平:慢阻肺非CSE组[(4.1±0.5)、(47.1±4.1)U/ml]显著低于健康非CSE组[(5.1±0.6)、(88.4 ±2.3)U/ml],慢阻肺CSE组和健康CSE组[(3.1±0.4)、(26.8±6.2) U/ml和(4.5±0.4)、(72.3±5.1) U/ml]分别显著低于慢阻肺非CSE组和健康非CSE组(均P＜0.01).丙二醛水平:慢阻肺非CSE组[(4.8 ±0.5)μmol/L]显著高于健康非CSE组[(2.1±0.4)μmol/L],慢阻肺CSE组和健康CSE组[(7.7±0.9)和(3.0±0.6) μmol/L]分别显著高于慢阻肺非CSE组和健康非CSE组(均P＜0.01).慢阻肺组平均荧光强度在基础状态下与TAC、GSH-PX水平呈正相关(r =0.523、0.818,P=0.038、0.001),与丙二醛水平呈负相关(r=-0.501,P=0.048),CSE干预后上述相关关系依然存在(r=0.704、0.716、-0.522,P=0.002、0.002、0.038).结论 香烟烟雾可导致慢性肺患者MDM吞噬能力下降,这可能与氧化应激密切相关.
目的 探討香煙煙霧提取物(CSE)對慢性阻塞性肺疾病(簡稱慢阻肺)患者單覈細胞源性巨噬細胞(MDM)吞噬功能的影響.方法 選擇2012年1月至2013年3月蘭州大學第一醫院慢阻肺穩定期患者32例,選擇同期健康體檢者32名,分離外週血單覈細胞,體外誘導培養成MDM.分彆將慢阻肺患者和健康體檢者的MDM分為慢阻肺非CSE組(常規培養)、慢阻肺CSE組(4％ CSE榦預6h)、健康非CSE組(常規培養)、健康CSE組(4％ CSE榦預6h).以流式細胞術的平均熒光彊度和激光共聚焦顯微鏡的熒光灰度值判斷MDM吞噬熒光標記大腸桿菌(FITC-E.coli的能力,菲囉啉比色法檢測細胞上清液總抗氧化能力(TAC),硫代巴比妥痠比色法檢測丙二醛含量,改良Hafeman直接測定法檢測穀胱甘肽過氧化物酶(GSH-PX)活力.結果 平均熒光彊度、熒光灰度值:慢阻肺非CSE組(20.2±2.2、51.5±5.8)顯著低于健康非CSE組(56.9±6.7、87.3 ±7.3),慢阻肺CSE組(7.6±0.7、14.1 ±3.4)和健康CSE組(48.0±5.4、69.7±6.0)分彆顯著低于慢阻肺非CSE組和健康非CSE組(均P＜0.01).TAC、GSH-PX水平:慢阻肺非CSE組[(4.1±0.5)、(47.1±4.1)U/ml]顯著低于健康非CSE組[(5.1±0.6)、(88.4 ±2.3)U/ml],慢阻肺CSE組和健康CSE組[(3.1±0.4)、(26.8±6.2) U/ml和(4.5±0.4)、(72.3±5.1) U/ml]分彆顯著低于慢阻肺非CSE組和健康非CSE組(均P＜0.01).丙二醛水平:慢阻肺非CSE組[(4.8 ±0.5)μmol/L]顯著高于健康非CSE組[(2.1±0.4)μmol/L],慢阻肺CSE組和健康CSE組[(7.7±0.9)和(3.0±0.6) μmol/L]分彆顯著高于慢阻肺非CSE組和健康非CSE組(均P＜0.01).慢阻肺組平均熒光彊度在基礎狀態下與TAC、GSH-PX水平呈正相關(r =0.523、0.818,P=0.038、0.001),與丙二醛水平呈負相關(r=-0.501,P=0.048),CSE榦預後上述相關關繫依然存在(r=0.704、0.716、-0.522,P=0.002、0.002、0.038).結論 香煙煙霧可導緻慢性肺患者MDM吞噬能力下降,這可能與氧化應激密切相關.
목적 탐토향연연무제취물(CSE)대만성조새성폐질병(간칭만조폐)환자단핵세포원성거서세포(MDM)탄서공능적영향.방법 선택2012년1월지2013년3월란주대학제일의원만조폐은정기환자32례,선택동기건강체검자32명,분리외주혈단핵세포,체외유도배양성MDM.분별장만조폐환자화건강체검자적MDM분위만조폐비CSE조(상규배양)、만조폐CSE조(4％ CSE간예6h)、건강비CSE조(상규배양)、건강CSE조(4％ CSE간예6h).이류식세포술적평균형광강도화격광공취초현미경적형광회도치판단MDM탄서형광표기대장간균(FITC-E.coli적능력,비라람비색법검측세포상청액총항양화능력(TAC),류대파비타산비색법검측병이철함량,개량Hafeman직접측정법검측곡광감태과양화물매(GSH-PX)활력.결과 평균형광강도、형광회도치:만조폐비CSE조(20.2±2.2、51.5±5.8)현저저우건강비CSE조(56.9±6.7、87.3 ±7.3),만조폐CSE조(7.6±0.7、14.1 ±3.4)화건강CSE조(48.0±5.4、69.7±6.0)분별현저저우만조폐비CSE조화건강비CSE조(균P＜0.01).TAC、GSH-PX수평:만조폐비CSE조[(4.1±0.5)、(47.1±4.1)U/ml]현저저우건강비CSE조[(5.1±0.6)、(88.4 ±2.3)U/ml],만조폐CSE조화건강CSE조[(3.1±0.4)、(26.8±6.2) U/ml화(4.5±0.4)、(72.3±5.1) U/ml]분별현저저우만조폐비CSE조화건강비CSE조(균P＜0.01).병이철수평:만조폐비CSE조[(4.8 ±0.5)μmol/L]현저고우건강비CSE조[(2.1±0.4)μmol/L],만조폐CSE조화건강CSE조[(7.7±0.9)화(3.0±0.6) μmol/L]분별현저고우만조폐비CSE조화건강비CSE조(균P＜0.01).만조폐조평균형광강도재기출상태하여TAC、GSH-PX수평정정상관(r =0.523、0.818,P=0.038、0.001),여병이철수평정부상관(r=-0.501,P=0.048),CSE간예후상술상관관계의연존재(r=0.704、0.716、-0.522,P=0.002、0.002、0.038).결론 향연연무가도치만성폐환자MDM탄서능력하강,저가능여양화응격밀절상관.
Objective To explore the effects of cigarette smoke extract (CSE) on phagocytosizing function of monocyte-derived macrophages (MDMs) in patients with chronic obstructive pulmonary disease (COPD).Methods From January 2012 to March 2013,peripheral blood monocytes were isolated from 32 stable COPD patients and 32 healthy controls at First Hospital,Lanzhou University.MDM was induced and cultured from monocytes in vitro.The MDMs from COPD patients and healthy controls were divided into 4 groups of COPD non-CSE (conventional culture),COPD CSE (4％ CSE treatment for 6 h),healthy nonCSE (conventional culture) and healthy CSE (4％ CSE treatment for 6 h).Flow cytometry (mean fluorescence intensity,MFI) and laser scanning confocal microscopy (fluorescence grey level) were applied to detect the ability of MDM phagocytosed fluorescein-labeled Escherichia coli (FITC-E.coli).Total antioxidative capacity (TAC) was measured by o-phenanthroline colorimetry.Malondialdehyde (MDA) was measured by thiobarbituricacid colorimetry and glutathione peroxidase (GSH-PX) by 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) method.Results MFI and fluorescence grey level in COPD non-CSE group (20.2 ± 2.2,51.5 ± 5.8) significantly decreased than those in healthy non-CSE group (56.9 ± 6.7,87.3 ±7.3).And in COPD CSE (7.6 ±0.7,14.1 ±3.4) and healthy CSE groups (48.0 ±5.4,69.7 ±6.0) decreased more than those in COPD non-CSE and healthy non-CSE groups (all P ＜ 0.01).The levels of TAC and GSH-PX in COPD non-CSE group ((4.1 ±0.5),(47.1 ±4.1) U/ml) were lower than those in healthy non-CSE group ((5.1 ± 0.6),(88.4 ± 2.3) U/ml).And in COPD CSE and healthy CSE groups ((3.1 ± 0.4),(26.8 ± 6.2) U/ml) and (4.5 ± 0.4),(72.3 ± 5.1) U/ml) were respectively lower than those in COPD non-CSE and healthy non-CSE groups (all P ＜ 0.01).The content of MDA in COPD non-CSE group was higher than that in healthy non-CSE group [(4.8 ±0.5) vs (2.1 ±0.4) μmol/L)].And in COPD CSE and healthy CSE groups ((7.7 ±0.9),(3.0 ±0.6) μmol/L) were higher than those in COPD non-CSE and healthy non-CSE groups (all P ＜ 0.01).At basic status,positive correlations existed between MFI and TAC,GSH-PX (r =0.523,0.818,P =0.038,0.001) while negative correlations between MFI and MDA (r =-0.501,P =0.048) in COPD patients and after CSE treatment,the above relationships still existed (r =0.704,0.716,-0.522,P =0.002,0.002,0.038).Conclusions Cigarette smoke can reduce the phagocytosizing ability of MDM in COPD patients.And it may be related with oxidative stress.
