丝裂霉素C及紫杉醇对人胚肺成纤维细胞增殖及凋亡的影响  

The influence of mitomycin C and paclitaxel on the proliferation and apoptosis of human pulmonary fibroblast

作　　者：陈楠[1] 张杰[1] 徐敏[1] 王婷[1] 王玉玲[1] 裴迎华[1] 

机构地区：[1]首都医科大学附属北京天坛医院呼吸内科,100050

出　　处：《中华结核和呼吸杂志》2013年第9期655-660,共6页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：北京市自然科学基金面上项目（7102053）

摘　　要：目的在体外细胞学水平观察丝裂霉素c及紫杉醇抑制人胚肺成纤维细胞增殖的量效及时效关系特点。初步探讨药物抑制细胞增殖的潜在机制，为设定药物洗脱气道支架的药物洗脱浓度提供试验参考。方法采用噻唑兰（MTT）法分别测定10-11mol／L至10-8moL／L（10倍倍比稀释）的丝裂霉素C或紫杉醇持续作用24、48和72h后对人胚肺戍纤维细胞增殖的抑制率。通过AnnexinV—FITC／PI双染及流式细胞术检测5×10-6、10-5、5×10-5、10-4、2×10-4mol／L的丝裂霉素C或紫杉醇持续作用48h后的细胞凋亡百分比。并采用Hoechst33342荧光染色法观察细胞凋亡的形态学特征。结果MTT结果显示，各浓度丝裂霉素C或紫杉醇持续作用24、48及72h均可在不同程度上抑制人胚肺成纤维细胞增殖。其中，当丝裂霉素C处于10-11mol／L至10-8mol／L的较低浓度水平时，药物对细胞增殖的抑制作用较弱。且在此浓度范围内，提高药物浓度或延长作用时间对于改善抑制率无益。而当丝裂霉素C处于10-7mol／L至10-4mol／L的较高浓度水平时，随作用时间延长或药物浓度提高，抑制率呈渐进式增高。10-7、10-6、10-5及10-4mol／L的丝裂霉素C持续作用72h，抑制率分别为53．52％、60．23％、89．81％及96．47％。紫杉醇对人胚肺成纤维细胞增殖的抑制作用存在明显的“阈浓度效应”。10-5mol／L的紫杉醇持续作用72h，抑制率仅为48．22％，但当浓度达到10-4mol／L后，药物作用24h时抑制率即可达93．38％，且随作用时间延长，抑制率可进一步升高。细胞凋亡部分的试验结果与Mr丌部分相吻合，即当药物对细胞增殖的抑制作用较为明显时，采用流式细胞术可检测到大量凋亡细胞，且以早期凋亡为主。此时进行Hoechst33342荧光染色可观察到典型的凋亡细胞。结论在体外条件下，一定浓度的丝裂霉素C或紫杉醇持续作用均可对人胚肺成�Objective To observe the inhibitory effect and potential mechanism of mitomycin C and paclitaxel on the proliferation of Human Puhnonary Fibroblast in vitro. So as to providing an experimental reference for the design of drug eluting airway stents. Methods Cell viability was measured by MTT assay after different concentrations of mitomycin C or paclitaxel varying from 10-11 mol/L to 10-4 mol/L had been applied to the fibroblasts for 24,48 or 72 b,respectively. Cell apoptosis was assessed by flow cytometry using dual staining with annexin V-FITC and propidium iodide 48 h after administering mitomycin C or paclitaxel at a concentration of 5× 10 -6,10 5,5 × 10 -5,10-4,2 × 10-4 mol/L, respectively. And the morphological character of cell apoptosis was observed by Hoechst 33342 fluorescent staining. Results The results of MTT revealed that cell proliferation were inhibited by mitomycin C and paclitaxel at all concentrations and exposure times. Among them, the inhibitory effect of mitomycin C were weak when the concentrations were between 10-11 mol/L to 10-5 mol/L. And within this context, the inhibitory ratio didn＇t correspond to the elevation of the concentration or the prolongation of the exposure times. However, when the concentration were between 10-7 mol/L to 10-4 tool/L, the inhibitory ratio rise progressively as the elevation of the concentration at all exposure times. The inhibitory ratio were 53.52% ,60. 23%, 89.81% and 96.47% respectively when cells were treated by 10 7,10 -6,10 -5 mol/L and 10-4 mol/L mitomycin C for 72 h. An apparent ＂threshold dose effect＂ was observed in the paclitaxel treated groups. It＇ s worth noting that the inhibitory ratio was only 48.22% when the cells had already been treated by 10-5 mol/L paclitaxel for 72 h. However, when the concentration had reached 10-4 mol/L, the inhibitory ratio sharply climbed to 93.38% even the cells had only been treated for 24 h. And the inhibitory ratio continued to rise as time prolonged. The results of cell apoptosis were consistent with

关 键 词：肉芽组织 成纤维细胞 丝裂霉素C 紫杉醇 药物洗脱气道支架 

分 类 号：R56[医药卫生—呼吸系统]