中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2012年
3期
131-136
,共6页
张翠萍%马筱玲%常文娇%周馨
張翠萍%馬篠玲%常文嬌%週馨
장취평%마소령%상문교%주형
葡萄球菌,金黄色%中性白细胞%肺炎%疾病模型,动物
葡萄毬菌,金黃色%中性白細胞%肺炎%疾病模型,動物
포도구균,금황색%중성백세포%폐염%질병모형,동물
Staphylococcusaureus%Neutrophils%Pneumonia%Disease models,animal
目的 探讨多形核粒细胞(PMN)在金黄色葡萄球菌Panton Valentine杀白细胞素(PVL)介导的肺炎性损伤中的作用.方法 取15只新西兰大白兔均分为3组,构建肺炎性损伤模型.对照组使用PBS灌肺,粒细胞正常兔直接用重组PVL灌肺(rPVL组),粒细胞减少症兔先用长春新碱(VCR)处理,再用rPVL灌肺(VCR+rPVL组).造模后9h,计数外周血和支气管肺泡灌洗液(BALF)中PMN,同时检测BALF上清液中乳酸脱氢酶(LDH)含量、肺通透指数(LPI),BALF中PMN凋亡率、坏死率、活性氧自由基(ROS)释放量.取肺组织检测肺湿干比,并进行病理学检查.组间比较采用t检验.结果 rPVL组外周血PMN为(2.69±0.34)×106/mL,显著低于对照组的(3.63±0.38)×106/mL(t=4.12,P＜0.05).对照组、rPVL组和VCR+rPVL组BALF中PMN分别为(0.57±0.01)×106/mL、(3.01±0.02)×106/mL和(0.10±0.02)×106/mL,rPVL组显著高于对照组(t=254.39,P＜0.05).rPVL组兔LDH、LPI和肺湿重/干重比均显著高于对照组,但后者与VCR+rPVL组比较,差异无统计学意义.rPVL组BALF中PMN晚期凋亡率和坏死率分别为( 18.98±1.04)％、(63.56±3.53)％,对照组分别为(1.03±0.17)％、(0.95± 0.33)％(t=38.24,39.48；均P＜0.05),VCR+rPVL组凋亡率和坏死率分别为(1.17=0.24)％、(1.1 3±0.17)％.rPVL组、对照组和VCR+rPVL组ROS分别为1.56±0.39、0.41±0.03和0.39±0.02,rPVL组明显升高(t=6.58,P＜0.05).rPVL组肺组织有弥漫性炎性细胞浸润、出血、水肿,VCR+rPVL组仅见支气管周围与肺泡间隔极少量炎性细胞浸润.结论 rPVL可引起粒细胞正常兔肺炎性损伤,但对粒细胞减少症兔肺损伤极小.PVL引起肺炎性损伤可能是依赖PMN的招募和聚集,继而坏死和(或)激活,释放细胞毒素颗粒内容物和(或)活性氧代谢产物等.
目的 探討多形覈粒細胞(PMN)在金黃色葡萄毬菌Panton Valentine殺白細胞素(PVL)介導的肺炎性損傷中的作用.方法 取15隻新西蘭大白兔均分為3組,構建肺炎性損傷模型.對照組使用PBS灌肺,粒細胞正常兔直接用重組PVL灌肺(rPVL組),粒細胞減少癥兔先用長春新堿(VCR)處理,再用rPVL灌肺(VCR+rPVL組).造模後9h,計數外週血和支氣管肺泡灌洗液(BALF)中PMN,同時檢測BALF上清液中乳痠脫氫酶(LDH)含量、肺通透指數(LPI),BALF中PMN凋亡率、壞死率、活性氧自由基(ROS)釋放量.取肺組織檢測肺濕榦比,併進行病理學檢查.組間比較採用t檢驗.結果 rPVL組外週血PMN為(2.69±0.34)×106/mL,顯著低于對照組的(3.63±0.38)×106/mL(t=4.12,P＜0.05).對照組、rPVL組和VCR+rPVL組BALF中PMN分彆為(0.57±0.01)×106/mL、(3.01±0.02)×106/mL和(0.10±0.02)×106/mL,rPVL組顯著高于對照組(t=254.39,P＜0.05).rPVL組兔LDH、LPI和肺濕重/榦重比均顯著高于對照組,但後者與VCR+rPVL組比較,差異無統計學意義.rPVL組BALF中PMN晚期凋亡率和壞死率分彆為( 18.98±1.04)％、(63.56±3.53)％,對照組分彆為(1.03±0.17)％、(0.95± 0.33)％(t=38.24,39.48；均P＜0.05),VCR+rPVL組凋亡率和壞死率分彆為(1.17=0.24)％、(1.1 3±0.17)％.rPVL組、對照組和VCR+rPVL組ROS分彆為1.56±0.39、0.41±0.03和0.39±0.02,rPVL組明顯升高(t=6.58,P＜0.05).rPVL組肺組織有瀰漫性炎性細胞浸潤、齣血、水腫,VCR+rPVL組僅見支氣管週圍與肺泡間隔極少量炎性細胞浸潤.結論 rPVL可引起粒細胞正常兔肺炎性損傷,但對粒細胞減少癥兔肺損傷極小.PVL引起肺炎性損傷可能是依賴PMN的招募和聚集,繼而壞死和(或)激活,釋放細胞毒素顆粒內容物和(或)活性氧代謝產物等.
목적 탐토다형핵립세포(PMN)재금황색포도구균Panton Valentine살백세포소(PVL)개도적폐염성손상중적작용.방법 취15지신서란대백토균분위3조,구건폐염성손상모형.대조조사용PBS관폐,립세포정상토직접용중조PVL관폐(rPVL조),립세포감소증토선용장춘신감(VCR)처리,재용rPVL관폐(VCR+rPVL조).조모후9h,계수외주혈화지기관폐포관세액(BALF)중PMN,동시검측BALF상청액중유산탈경매(LDH)함량、폐통투지수(LPI),BALF중PMN조망솔、배사솔、활성양자유기(ROS)석방량.취폐조직검측폐습간비,병진행병이학검사.조간비교채용t검험.결과 rPVL조외주혈PMN위(2.69±0.34)×106/mL,현저저우대조조적(3.63±0.38)×106/mL(t=4.12,P＜0.05).대조조、rPVL조화VCR+rPVL조BALF중PMN분별위(0.57±0.01)×106/mL、(3.01±0.02)×106/mL화(0.10±0.02)×106/mL,rPVL조현저고우대조조(t=254.39,P＜0.05).rPVL조토LDH、LPI화폐습중/간중비균현저고우대조조,단후자여VCR+rPVL조비교,차이무통계학의의.rPVL조BALF중PMN만기조망솔화배사솔분별위( 18.98±1.04)％、(63.56±3.53)％,대조조분별위(1.03±0.17)％、(0.95± 0.33)％(t=38.24,39.48；균P＜0.05),VCR+rPVL조조망솔화배사솔분별위(1.17=0.24)％、(1.1 3±0.17)％.rPVL조、대조조화VCR+rPVL조ROS분별위1.56±0.39、0.41±0.03화0.39±0.02,rPVL조명현승고(t=6.58,P＜0.05).rPVL조폐조직유미만성염성세포침윤、출혈、수종,VCR+rPVL조부견지기관주위여폐포간격겁소량염성세포침윤.결론 rPVL가인기립세포정상토폐염성손상,단대립세포감소증토폐손상겁소.PVL인기폐염성손상가능시의뢰PMN적초모화취집,계이배사화(혹)격활,석방세포독소과립내용물화(혹)활성양대사산물등.
Objective To explore the role of polymorpbonuclear leukocyte (PMN) in PantonValentine leucocidin (PVL)-induccd acute lung inflammation and injury. Methods Fifteen New Zealand rabbits were divided into 3 groups with five rabbits in each group.The controls were treated with pbosphate buffer solution (PBS),the rabbits with normal granulocyte in rPVL group were treated with endotracheal instillation of rPVL,the granulocytopenia rabbits in vincristine (VCR) +rPVL group were firstly treated with VCR,thcn with endotracheal instillation of rPVL.Nine hours after injection,the peripheral blood and bronchoalveolar lavage fluid (BALF) were collected for counting PMN.The lactate dehydrogenase (LDH) activity in BALF,lung permeability index (LPI),PMN apoptosis and necrosis and the release of reactive oxygen species (ROS) in BALF were measured.After the rabbits sacrificed,the lung tissue samples were collcctcd for dctcrmining wet/dry (W/D) ratio and histopathological examination.The comparison among groups was done by t test.Results The PMN count in the peripheral blood was (2.69=0.34) × 10 mL in rPVL group,which was significantly lower than control group [(3.63 ± 0.38) × 105/mL] (t =4.12,P＜0.05).The PMN counts in BALF in control group,rPVL group and VCR+rPVL group were (0.57±0.01 ) ×106/mL,(3.01±0.02) × 106/mL and (0.10±0.02) × 106/mL,respectively; that in rPVL group was significantly higher than those in control group (t=254.39,P＜0.05).The LDH activity,LPI and W/D ratio in rPVL group were all significantly higher than control group,while those in VCR+rPVL group were not significantly different from control group.The PMN apoptosis rate and necrosis rate in VCR+rPVL groupwere (1.17±0.24)％ and (1.13±0.17)％,respectively.The releases of ROS (meanfluorescence intensity) in rPVL group,control group and VCR+rPVL group were 1.56±0.39,0.41±0.03 and 0.39±0.02,respectively,and that in rPVL group was significantly higher (t=6.58,P＜0.05).Histopathological examination of the lung showed the diffuse infiltration of inflammatory cells,hemorrhage and edema in rPVL group,wbile there was only thimbleful infiltration of inflammatory cells observed in surrounding bronchia and alveolar septun in VCR-rPVL group.Conclusions rPVL can induce lung inflammation and injury in rabbits with normal granulocyte,but not in neutropenic rabbirs.Lung inflammation and injury may be the result of recruitment,aggregation and subsequent lysis and/or activation of PMN,which can damage the lung by releasing the contents of cytotoxic granules and/or reactive oxygen metabolites.
