CAJ | 학술논문

目的探讨重组人粒细胞集落刺激因子(recombinant human granulocyte colony stimulating factor,rhG-CSF)对大鼠肺动脉高压的治疗效果及其对循环内皮祖细胞(endothelial progenitor cells,EPCs)数量及功能的影响. 方法 8周龄Sprague-Dawlay大鼠随机分为模型组、治疗组和对照组,每组8只.模型组及治疗组大鼠一次性皮下注射1％的野百合碱50 mg/kg以诱导形成肺动脉高压模型,对照组注射等量磷酸盐缓冲液.治疗组于第5天皮下注射rhG-CSF 50 μg/(kg·d),连续3d.各组大鼠分别于第21天经尾静脉取血,应用流式细胞仪计数每100 000个单个核细胞中EPCs所占比例.检测右心室收缩压,HE染色观察肺组织及肺血管病理变化.尾静脉采集的外周血单个核细胞进行EPCs体外培养,14d后采用四甲基偶氮唑蓝比色法、黏附实验和迁移实验测定EPCs的增殖、黏附和迁移能力.多组样本之间的比较采用单因素方差分析,组间两两比较采用LSD检验.结果 (1)肺动脉压力及肺小动脉病理改变:模型组大鼠右心室收缩压较对照组明显升高[(48.13±2.85) mm Hg与(27.88±3.04) mm Hg,t=2.016,P＜0.01],且肺小动脉血管内皮细胞损伤明显,管壁显著增厚.治疗组大鼠右心室收缩压[(30.38±2.83) mm Hg]较模型组明显降低,接近对照组水平(t=0.376,P＞0.05),且肺部病变也相应减轻.(2)循环EPCs占单个核细胞比例:模型组大鼠外周血EPCs占单个核细胞比例较对照组明显下降[(0.016±0.007)％与(0.031±0.011)％,t=2.617,P＜0.01].治疗后,大鼠循环EPCs占单个核细胞比例[(0.042±0.01 3)％]明显高于模型组(t=4.325,P＜0.01)及对照组(t=1.942,P＜0.05).(3)体外培养的EPCs的增殖、黏附、迁移能力:模型组EPCs细胞增殖力、黏附细胞数和迁移细胞数分别为0.49±0.04,(6.93±1.47)个/高倍视野和(7.22±1.53)个/高倍视野,均低于对照组[分别为0.68±0.07、(11.05±1.73)个/高倍视野和(12.58±2.15)个/高倍视野]和治疗组[0.63±0.06、(12.35±1.82)个/高倍视野和(12.97±2.84)个/高倍视野],差异均有统计学意义(P均＜0.05). 结论 rhG-CSF能够有效上调肺动脉高压大鼠循环EPCs的数量及功能,从而有助于肺动脉高压的防治. 目的探討重組人粒細胞集落刺激因子(recombinant human granulocyte colony stimulating factor,rhG-CSF)對大鼠肺動脈高壓的治療效果及其對循環內皮祖細胞(endothelial progenitor cells,EPCs)數量及功能的影響. 方法 8週齡Sprague-Dawlay大鼠隨機分為模型組、治療組和對照組,每組8隻.模型組及治療組大鼠一次性皮下註射1％的野百閤堿50 mg/kg以誘導形成肺動脈高壓模型,對照組註射等量燐痠鹽緩遲液.治療組于第5天皮下註射rhG-CSF 50 μg/(kg·d),連續3d.各組大鼠分彆于第21天經尾靜脈取血,應用流式細胞儀計數每100 000箇單箇覈細胞中EPCs所佔比例.檢測右心室收縮壓,HE染色觀察肺組織及肺血管病理變化.尾靜脈採集的外週血單箇覈細胞進行EPCs體外培養,14d後採用四甲基偶氮唑藍比色法、黏附實驗和遷移實驗測定EPCs的增殖、黏附和遷移能力.多組樣本之間的比較採用單因素方差分析,組間兩兩比較採用LSD檢驗.結果 (1)肺動脈壓力及肺小動脈病理改變:模型組大鼠右心室收縮壓較對照組明顯升高[(48.13±2.85) mm Hg與(27.88±3.04) mm Hg,t=2.016,P＜0.01],且肺小動脈血管內皮細胞損傷明顯,管壁顯著增厚.治療組大鼠右心室收縮壓[(30.38±2.83) mm Hg]較模型組明顯降低,接近對照組水平(t=0.376,P＞0.05),且肺部病變也相應減輕.(2)循環EPCs佔單箇覈細胞比例:模型組大鼠外週血EPCs佔單箇覈細胞比例較對照組明顯下降[(0.016±0.007)％與(0.031±0.011)％,t=2.617,P＜0.01].治療後,大鼠循環EPCs佔單箇覈細胞比例[(0.042±0.01 3)％]明顯高于模型組(t=4.325,P＜0.01)及對照組(t=1.942,P＜0.05).(3)體外培養的EPCs的增殖、黏附、遷移能力:模型組EPCs細胞增殖力、黏附細胞數和遷移細胞數分彆為0.49±0.04,(6.93±1.47)箇/高倍視野和(7.22±1.53)箇/高倍視野,均低于對照組[分彆為0.68±0.07、(11.05±1.73)箇/高倍視野和(12.58±2.15)箇/高倍視野]和治療組[0.63±0.06、(12.35±1.82)箇/高倍視野和(12.97±2.84)箇/高倍視野],差異均有統計學意義(P均＜0.05). 結論 rhG-CSF能夠有效上調肺動脈高壓大鼠循環EPCs的數量及功能,從而有助于肺動脈高壓的防治.
목적 탐토중조인립세포집락자격인자(recombinant human granulocyte colony stimulating factor,rhG-CSF)대대서폐동맥고압적치료효과급기대순배내피조세포(endothelial progenitor cells,EPCs)수량급공능적영향. 방법 8주령Sprague-Dawlay대서수궤분위모형조、치료조화대조조,매조8지.모형조급치료조대서일차성피하주사1％적야백합감50 mg/kg이유도형성폐동맥고압모형,대조조주사등량린산염완충액.치료조우제5천피하주사rhG-CSF 50 μg/(kg·d),련속3d.각조대서분별우제21천경미정맥취혈,응용류식세포의계수매100 000개단개핵세포중EPCs소점비례.검측우심실수축압,HE염색관찰폐조직급폐혈관병리변화.미정맥채집적외주혈단개핵세포진행EPCs체외배양,14d후채용사갑기우담서람비색법、점부실험화천이실험측정EPCs적증식、점부화천이능력.다조양본지간적비교채용단인소방차분석,조간량량비교채용LSD검험.결과 (1)폐동맥압력급폐소동맥병리개변:모형조대서우심실수축압교대조조명현승고[(48.13±2.85) mm Hg여(27.88±3.04) mm Hg,t=2.016,P＜0.01],차폐소동맥혈관내피세포손상명현,관벽현저증후.치료조대서우심실수축압[(30.38±2.83) mm Hg]교모형조명현강저,접근대조조수평(t=0.376,P＞0.05),차폐부병변야상응감경.(2)순배EPCs점단개핵세포비례:모형조대서외주혈EPCs점단개핵세포비례교대조조명현하강[(0.016±0.007)％여(0.031±0.011)％,t=2.617,P＜0.01].치료후,대서순배EPCs점단개핵세포비례[(0.042±0.01 3)％]명현고우모형조(t=4.325,P＜0.01)급대조조(t=1.942,P＜0.05).(3)체외배양적EPCs적증식、점부、천이능력:모형조EPCs세포증식력、점부세포수화천이세포수분별위0.49±0.04,(6.93±1.47)개/고배시야화(7.22±1.53)개/고배시야,균저우대조조[분별위0.68±0.07、(11.05±1.73)개/고배시야화(12.58±2.15)개/고배시야]화치료조[0.63±0.06、(12.35±1.82)개/고배시야화(12.97±2.84)개/고배시야],차이균유통계학의의(P균＜0.05). 결론 rhG-CSF능구유효상조폐동맥고압대서순배EPCs적수량급공능,종이유조우폐동맥고압적방치.
Objective To investigate the effects of recombinant human granulocyte colony stimulating factor (rhG-CSF) therapy on pulmonary hypertension,and its influence on number and functions of circulating endothelial progenitor cells (EPCs) in rats.Methods Eight week old Sprague-Dawlay rats were randomized into model group,treatment group and control group (8 rats in each group).The rats in model group and treatment group were treated with single subcutaneous injection of 1％ monocrotaline (50 mg/kg) to induce pulmonary hypertension models,while the control group was treated with phosphate buffered saline.Five days later,the rats in treatment group were administrated with 50 μg/(kg· d) rhG-CSF for 3 days.On day 21,peripheral blood was collected from caudal vein in all groups,and the percentage of EPCs in 100 000 mononuclear cells was evaluated by flow cytometry.Right ventricular systolic pressure was assessed,and the pathological changes of lung tissue and pneumoangiogram were observed by HE staining.Meanwhile,peripheral mononuclear cells collected from caudal vein were separated and cultured in vitro for EPCs.The cell ffunctions as proliferation,adhesion and migration ability were assessed.ANOVA and LSD test were applied as statistical analysis methods.Results (1) The right ventricular systolic pressure of rats in model group was higher than that in the controls [(48.13 ± 2.85) mm Hg vs (27.88 ± 3.04) mm Hg,t=2.016,P＜0.01],the lesion of endothelial cells in pulmonary arteriolar was evident,and the vessel wall was thickened.The pulmonary artery pressure of rats in the treatment group [(30.38 ± 2.83) mm Hg] was lower than that in the model group and close to the level of control group (t=0.376,P＞0.05) with mild pulmonary pathological changes.(2) The percentage off peripheral blood EPCs in mononuclear cells in the model group was decreased as compared to the control group [(0.016±0.007) ％ vs (0.031±0.011) ％,t=2.617,P＜0.01].After administration ofrhG-CSF,the EPCs in treatment group [(0.042±0.013) ％] was increased evidently as compared to the model group (t=4.325,P＜0.01) and the control group (t =1.942,P＜0.05).(3) The proliferation,adhesive and migrated cells of EPCs in model group were 0.49 ± 0.04,(6.93 ± 1.47) cells/HPF and (7.22±1.53) cells/HPF,lower than those in control group [0.68±0.07,(11.05±1.73) cells/HPF and (12.58±2.15) cells/HPF] and treatment group [0.63±0.06,(12.35±1.82) cells/HPF and (12.97±2.84) cells/HPF],the differences were statistically significant(all P＜0.05).Conclusions rhG-CSF may be effective in treating pulmonary hypertension through up-regulating the number and function of circulating EPCs in rat model of pulmonary hypertension.