中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
22期
1560-1564
,共5页
刘俊峰%杜忠东%陈植%李慎涛%路敦祥%李丽
劉俊峰%杜忠東%陳植%李慎濤%路敦祥%李麗
류준봉%두충동%진식%리신도%로돈상%리려
黏膜皮肤淋巴结综合征%模型,动物%干酪乳杆菌%内皮祖细胞
黏膜皮膚淋巴結綜閤徵%模型,動物%榦酪乳桿菌%內皮祖細胞
점막피부림파결종합정%모형,동물%간락유간균%내피조세포
Mucocutaneous lymph node syndrome%Model,animal%Lactobacillus casei%Endothelial progenitor cell
目的 探讨川崎病模型小鼠冠状动脉损伤的发生与内皮祖细胞(EPC)数量及功能改变的相关性.方法 制备酪乳杆菌细胞壁提取物( LCWE),取0.5 ml经腹腔单次注射制备C57BL/6小鼠的川崎病模型.将24只小鼠以随机数字表法分为3组:LCWE注射14 d组；LCWE注射56 d组及对照组(磷酸盐缓冲液腹腔注射),每组8只.分别检测其外周血CD34+ Flk-1+ CD45 -的EPC含量,同时提取各组小鼠的骨髓单个核细胞在体外进行EPC培养,培养至第7天,通过噻唑蓝(MTT)比色法、黏附实验和迁移实验分别测定EPC的增殖、黏附和迁移能力.结果 LCWE注射14 d组小鼠冠状动脉主干及其各级分支周围可以见到以淋巴细胞为主的大量炎细胞浸润,其外周血中循环EPC的含量(0.017％±0.008％)明显低于对照组(0.028％ ±0.007％),LCWE注射56 d组冠状动脉弹力层的破坏清晰可见,而其外周血循环EPC的含量(0.016％±0.007％)也明显低于对照组(均P＜0.01).通过骨髓单个核细胞的体外培养显示,LCWE注射14 d组小鼠骨髓EPC的集落形成能力明显低于对照组,其体外增殖能力检测的MTT实验中吸光度(A值)为0.39±0.11,低于对照组(0.61±0.14,P＜0.01)；其黏附能力和迁移能力[(3.1±0.6)、(3.2±0.6)个/高倍视野)]也均低于对照组[(6.4±1.2)、(6.2±0.5)个/高倍视野,均P＜0.01)].LCWE注射56 d组小鼠骨髓EPC集落形成能力仍明显低于对照组,其增殖能力、黏附能力和迁移能力[0.38±0.09、(3.1±0.6)个/高倍视野和(3.3±0.6)个/高倍视野]也显著低于对照组(均P＜0.01).结论 川崎病模型小鼠冠状动脉损伤的发生可能与EPC数量及功能的下调存在一定的相关性.
目的 探討川崎病模型小鼠冠狀動脈損傷的髮生與內皮祖細胞(EPC)數量及功能改變的相關性.方法 製備酪乳桿菌細胞壁提取物( LCWE),取0.5 ml經腹腔單次註射製備C57BL/6小鼠的川崎病模型.將24隻小鼠以隨機數字錶法分為3組:LCWE註射14 d組；LCWE註射56 d組及對照組(燐痠鹽緩遲液腹腔註射),每組8隻.分彆檢測其外週血CD34+ Flk-1+ CD45 -的EPC含量,同時提取各組小鼠的骨髓單箇覈細胞在體外進行EPC培養,培養至第7天,通過噻唑藍(MTT)比色法、黏附實驗和遷移實驗分彆測定EPC的增殖、黏附和遷移能力.結果 LCWE註射14 d組小鼠冠狀動脈主榦及其各級分支週圍可以見到以淋巴細胞為主的大量炎細胞浸潤,其外週血中循環EPC的含量(0.017％±0.008％)明顯低于對照組(0.028％ ±0.007％),LCWE註射56 d組冠狀動脈彈力層的破壞清晰可見,而其外週血循環EPC的含量(0.016％±0.007％)也明顯低于對照組(均P＜0.01).通過骨髓單箇覈細胞的體外培養顯示,LCWE註射14 d組小鼠骨髓EPC的集落形成能力明顯低于對照組,其體外增殖能力檢測的MTT實驗中吸光度(A值)為0.39±0.11,低于對照組(0.61±0.14,P＜0.01)；其黏附能力和遷移能力[(3.1±0.6)、(3.2±0.6)箇/高倍視野)]也均低于對照組[(6.4±1.2)、(6.2±0.5)箇/高倍視野,均P＜0.01)].LCWE註射56 d組小鼠骨髓EPC集落形成能力仍明顯低于對照組,其增殖能力、黏附能力和遷移能力[0.38±0.09、(3.1±0.6)箇/高倍視野和(3.3±0.6)箇/高倍視野]也顯著低于對照組(均P＜0.01).結論 川崎病模型小鼠冠狀動脈損傷的髮生可能與EPC數量及功能的下調存在一定的相關性.
목적 탐토천기병모형소서관상동맥손상적발생여내피조세포(EPC)수량급공능개변적상관성.방법 제비락유간균세포벽제취물( LCWE),취0.5 ml경복강단차주사제비C57BL/6소서적천기병모형.장24지소서이수궤수자표법분위3조:LCWE주사14 d조；LCWE주사56 d조급대조조(린산염완충액복강주사),매조8지.분별검측기외주혈CD34+ Flk-1+ CD45 -적EPC함량,동시제취각조소서적골수단개핵세포재체외진행EPC배양,배양지제7천,통과새서람(MTT)비색법、점부실험화천이실험분별측정EPC적증식、점부화천이능력.결과 LCWE주사14 d조소서관상동맥주간급기각급분지주위가이견도이림파세포위주적대량염세포침윤,기외주혈중순배EPC적함량(0.017％±0.008％)명현저우대조조(0.028％ ±0.007％),LCWE주사56 d조관상동맥탄력층적파배청석가견,이기외주혈순배EPC적함량(0.016％±0.007％)야명현저우대조조(균P＜0.01).통과골수단개핵세포적체외배양현시,LCWE주사14 d조소서골수EPC적집락형성능력명현저우대조조,기체외증식능력검측적MTT실험중흡광도(A치)위0.39±0.11,저우대조조(0.61±0.14,P＜0.01)；기점부능력화천이능력[(3.1±0.6)、(3.2±0.6)개/고배시야)]야균저우대조조[(6.4±1.2)、(6.2±0.5)개/고배시야,균P＜0.01)].LCWE주사56 d조소서골수EPC집락형성능력잉명현저우대조조,기증식능력、점부능력화천이능력[0.38±0.09、(3.1±0.6)개/고배시야화(3.3±0.6)개/고배시야]야현저저우대조조(균P＜0.01).결론 천기병모형소서관상동맥손상적발생가능여EPC수량급공능적하조존재일정적상관성.
Objective To assess whether the occurrence of coronary artery lesion was correlated with the changes of endothelial progenitor cell (EPC) number and function in murine model of Kawasaki disease (KD).Methods Lactobacillus casei cell wall extract (LCWE) was prepared and then C57BL/6 mice received a single intraperitoneal injection of LCWE for inducing KD.Twenty-four mice were categorized randomly into 3 groups:KD model group at Day 14 post-injection,KD model group at Day 56 post-injection and control group with an intraperitoneal injection of phosphate buffered solution (n =8 each).The number of circulating EPC was defined as CD34 + Flk-1 + CD45 - from mice.Meanwhile,bone warrow mononuclear cells were cultured in vitro to expand EPC for functional analysis. After 7 days of culturing,EPC were inoculated onto culture plate and thiazolyl blue assay was used to measure the absorbance value by enzyme labeling instrument to evaluate the proliferation.The adhesion of EPC was performed by replating cells on fibronectin coated dishes and then counting the number of adherent cells.The migration of EPC was assayed by Transwell.Results Focal inflammatory infiltrate was evident in coronary artery trunk and a series of branches at Day 14 post-injection.The inflammatory cell infiltrate consisted of mononuclear lymphocytes.The number of circulating EPC were significantly lower in the Day 14 LCWE-treating murine model versus the controls (0.017％ ±0.008％ vs 0.028％ ±0.007％,P ＜0.01 ).Disruption of elastin was consistently observed at Day 56 post-injection.And there was no apparent recovery in number of EPC (0.016％ ±0.007％,P ＜ 0.01 ). When bone marrow mononuclear cells were cultured in vitro,the colony-forming ability of EPC decreased in the KD model group at Day 14 post-injection versus the controls.Test of proliferating ability showed that the absorbance was 0.39 ±0.11 in MTT experiment and decreased than the controls (0.61 ±0.14,P ＜ 0.01 ).Adhesion and migration were also down-regulated versus the controls ((3.1 ±:0.6) and (3.2±0.6) vs (6.4±1.2) and (6.2±0.5) cells/HPF,both P＜0.01).In the KD model group at Day 56 post-injection,the colony-forming ability of EPC was not recovered significantly.Proliferation ability,adhesion and migration were still decreased compared to the controls (0.38 ±0.09,(3.12 ±0.56) cells/HPF and (3.29 ±0.63 ) cells/HPF,all P ＜ 0.01 ).Conclusion The occurrence of coronary artery lesion may be correlated with the down-regulation of EPC number and function in murine model of KD.
