大鼠远端肺动脉平滑肌细胞原代培养及其生物学特性和功能的研究

发布时间：2018-06-21 06:00

本文选题：肌细胞 + 平滑肌　；参考：《中国全科医学》2014年27期

【摘要】：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,P0.05)。COPD模型组PASMCs增殖(1.315±0.203)强于对照组(1.023±0.057)(t=3.387,P0.05)。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。
[Abstract]:Objective by tissue adherent culture of primary rat distal pulmonary artery smooth muscle cells (PASMCs), chronic obstructive pulmonary disease (COPD) and the biological characteristics and function of model rat and normal rat distal PASMCs. Methods SPF SD male rats 12, according to the random number table method divided into 2 groups: control group rats 1,14 days Intratracheal injection of 0.9% Sodium Chloride Solution, no cigarette smoke exposure of.COPD rats in the 1,14 days after injection of lipopolysaccharide in the airway (LPS) solution, 200 g/, 1 h/d of cigarette smoke exposure, a total of 8 weeks. After the establishment of the model, the cell morphology was observed by tissue adherent extraction and primary cultured rats the distal PASMCs. under inverted phase contrast microscope And the growth characteristics of cells; immunohistochemical staining, transmission electron microscopy and fluorescence identification of PASMCs CCK-8 cells; flow cytometry was used to detect the proliferation of cells in each group. Results under inverted phase contrast microscope PASMCs were fusiform or fishtail, growth is the typical "peak and valley" pattern; immunohistochemical staining, transmission electron microscopy identified above the cells for 97% PA The SMCs.COPD model group, PASMCs absorbance (1.079 + 0.119) higher than that of the control group (0.871 + 0.041) (t=4.039, P0.05).COPD model group PASMCs proliferation (1.315 + 0.203) than in control group (1.023 + 0.057) (t=3.387, P0.05). Conclusion PASMCs morphology control group and COPD model group in primary culture is similar, but the biological characteristics of different functions, there are also differences.
【作者单位】：桂林医学院附属医院呼吸内科;
【基金】：国家自然科学基金项目(81360010) 广西壮族自治区卫生厅项目(重2012004)
【分类号】：R563.9

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