中国全科医学
中國全科醫學
중국전과의학
CHINESE GENERAL PRACTICE
2014年
27期
3207-3210
,共4页
廖运学%王昌明%蒋明%马礼兵%徐青%陈峰%吕倩
廖運學%王昌明%蔣明%馬禮兵%徐青%陳峰%呂倩
료운학%왕창명%장명%마례병%서청%진봉%려천
肌细胞,平滑肌%肺动脉%大鼠%肺疾病,慢性阻塞性
肌細胞,平滑肌%肺動脈%大鼠%肺疾病,慢性阻塞性
기세포,평활기%폐동맥%대서%폐질병,만성조새성
Myocytes,smooth muscle%Pulmonary artery%Rats%Pulmonary disease,chronic obstructive
目的:采用组织块贴壁法体外培养原代大鼠远端肺动脉平滑肌细胞( PASMCs),探讨慢性阻塞性肺疾病(COPD)模型大鼠与正常大鼠远端PASMCs的生物学特性及功能的异同。方法 SPF级SD雄性大鼠12只,按随机数字表法分为2组:对照组大鼠第1、14天经气道内注入0.9%氯化钠溶液,无香烟烟雾暴露。COPD模型组大鼠于实验第1、14天经气道内注入脂多糖( LPS)溶液,200μg/次,香烟烟雾暴露1 h/d,共8周。模型建立后,组织块贴壁法提取并原代培养各组大鼠远端PASMCs。倒置相差显微镜下观察细胞形态及生长特点;细胞免疫组化染色、透射电镜鉴定PASMCs;CCK-8和细胞荧光流式细胞仪检测各组细胞增殖情况。结果倒置相差显微镜下PASMCs为长梭形或鱼尾状,为典型的“峰-谷”样生长;细胞免疫组化染色、透射电镜鉴定所培养的细胞97%以上为PASMCs。COPD模型组PASMCs吸光度(1.079±0.119)高于对照组(0.871±0.041)(t=4.039,P<0.05)。COPD模型组PASMCs增殖(1.315±0.203)强于对照组(1.023±0.057)(t=3.387,P<0.05)。结论对照组与COPD模型组原代培养的PASMCs形态学相似,但生物学特性不同,功能亦存在差异。
目的:採用組織塊貼壁法體外培養原代大鼠遠耑肺動脈平滑肌細胞( PASMCs),探討慢性阻塞性肺疾病(COPD)模型大鼠與正常大鼠遠耑PASMCs的生物學特性及功能的異同。方法 SPF級SD雄性大鼠12隻,按隨機數字錶法分為2組:對照組大鼠第1、14天經氣道內註入0.9%氯化鈉溶液,無香煙煙霧暴露。COPD模型組大鼠于實驗第1、14天經氣道內註入脂多糖( LPS)溶液,200μg/次,香煙煙霧暴露1 h/d,共8週。模型建立後,組織塊貼壁法提取併原代培養各組大鼠遠耑PASMCs。倒置相差顯微鏡下觀察細胞形態及生長特點;細胞免疫組化染色、透射電鏡鑒定PASMCs;CCK-8和細胞熒光流式細胞儀檢測各組細胞增殖情況。結果倒置相差顯微鏡下PASMCs為長梭形或魚尾狀,為典型的“峰-穀”樣生長;細胞免疫組化染色、透射電鏡鑒定所培養的細胞97%以上為PASMCs。COPD模型組PASMCs吸光度(1.079±0.119)高于對照組(0.871±0.041)(t=4.039,P<0.05)。COPD模型組PASMCs增殖(1.315±0.203)彊于對照組(1.023±0.057)(t=3.387,P<0.05)。結論對照組與COPD模型組原代培養的PASMCs形態學相似,但生物學特性不同,功能亦存在差異。
목적:채용조직괴첩벽법체외배양원대대서원단폐동맥평활기세포( PASMCs),탐토만성조새성폐질병(COPD)모형대서여정상대서원단PASMCs적생물학특성급공능적이동。방법 SPF급SD웅성대서12지,안수궤수자표법분위2조:대조조대서제1、14천경기도내주입0.9%록화납용액,무향연연무폭로。COPD모형조대서우실험제1、14천경기도내주입지다당( LPS)용액,200μg/차,향연연무폭로1 h/d,공8주。모형건립후,조직괴첩벽법제취병원대배양각조대서원단PASMCs。도치상차현미경하관찰세포형태급생장특점;세포면역조화염색、투사전경감정PASMCs;CCK-8화세포형광류식세포의검측각조세포증식정황。결과도치상차현미경하PASMCs위장사형혹어미상,위전형적“봉-곡”양생장;세포면역조화염색、투사전경감정소배양적세포97%이상위PASMCs。COPD모형조PASMCs흡광도(1.079±0.119)고우대조조(0.871±0.041)(t=4.039,P<0.05)。COPD모형조PASMCs증식(1.315±0.203)강우대조조(1.023±0.057)(t=3.387,P<0.05)。결론대조조여COPD모형조원대배양적PASMCs형태학상사,단생물학특성불동,공능역존재차이。
Objective Toinvestigatethesimilaritiesanddifferencesofbiologicalcharacteristicsandfunctionsbetween chronic obstructive pulmonary disease ( COPD ) model rats and normal rats distal pulmonary arterial smooth muscle cells (PASMCs)byusingtissueadherentculturedPASMCs.Methods 12SPFSDratswererandomlydividedintotwogroups:the control group was given saline injection on the 1st and 14th day,without exposure to cigarette smoke;The COPD model group was given intratracheal injection of lipopolysaccharide( LPS)solution,200μg/time,as well as cigarette smoke exposure of 1 h/d for a total of eight weeks. After the model was established,tissue adherent method was used to culture distal PASMCs. Cell mor-phology and growth characteristics were observed under inverted phase contrast microscope. Cell immunohistochemistry and elec-tron microscopy was used to identify PASMCs,and CCK-8 and fluorescence flow cytometry was used to detect cell prolifera-tion.Results Underinvertedphasecontrastmicroscope,PASMCsshapedlikelongspindleorfishtail,typically"peak-val-ley" style growth;Cell immunohistochemistry and electron microscopy identified that 97% of the cultured cells were PASMCs. The PASMCs absorbance of the COPD model group(1. 079±0. 119)was higher than that of the control group(0. 871±0. 041)(t=4. 039,P<0. 05). The PASMCs proliferation of COPD model group(1. 315±0. 203)was stronger than that of thecontrolgroup(1.023±0.057)(t=3.387,P<0.05).℅onclusion TheprimaryculturedPASMCshavesimilarmorphol-ogy in the control group and COPD model group,but have different biological characteristics and functionality.