毒胡萝卜素对转化生长因子β_1刺激的小鼠肺成纤维细胞凋亡的影响  

Effects of thapsigargin on the apoptosis in mouse lung fibroblast stimulated by transforming growth factor-β_1

作　　者：闫宇 马思琪 肖倩[2] 曲银娥 肖永红 

机构地区：[1]华北理工大学临床医学院,河北唐山063000 [2]华北理工大学冀唐学院,河北唐山063000 [3]华北理工大学基础医学院,河北唐山063000 [4]华北理工大学公共卫生学院,河北唐山063000

出　　处：《中国临床药理学杂志》2017年第19期1915-1917,共3页The Chinese Journal of Clinical Pharmacology

基　　金：河北省科技厅科学技术研究与发展计划基金资助项目(10276113D);华北理工大学大学生创新创业训练计划基金资助项目(X2017045)

摘　　要：目的研究毒胡萝卜素对转化生长因子β_1(TGF-β_1)刺激的小鼠肺成纤维细胞周期、细胞凋亡及半胱氨酸特异性蛋白酶-12(Caspase-12)蛋白表达的影响。方法液氮冻存的小鼠肺成纤维细胞株(L929),经复苏传代培养,同步化后分为3组:对照组、模型组和实验组,对照组的L929细胞只加入RPMI-1640培养基,模型组为L929+5 ng·m L^(-1)TGF-β_1,实验组为L929+5ng·m L^(-1)TGF-β_1诱导24 h+4μmol·L^(-1)毒胡萝卜素干预24 h。用流式细胞仪检测细胞周期和细胞凋亡情况,以免疫组织细胞化学方法检测Caspase-12蛋白表达。结果细胞周期:各组间的G1期、S期、G2期变化差异均有统计学意义(均P<0.05)。对照组、模型组和实验组的G1期细胞比例分别为(58.72±8.35)%,(52.13±7.61)%,(73.02±9.32)%,这3组的S期比例分别为(29.50±4.24)%,(40.54±6.31)%,(21.66±4.17)%,这3组的G2期比例分别为(11.75±0.96)%,(7.62±3.22)%,(6.09±2.11)%,组间比较差异均有统计学意义(均P<0.05);但G2期实验组和模型组比较差异无统计学意义(P>0.05);对照组、模型组、实验组的凋亡率分别为6.73%,5.03%,21.53%,caspase-12蛋白表达分别为3356.45±205.74,2645.56±168.36,5424.76±241.79,这2项指标组间比较差异均有统计学意义(均P<0.05)。结论毒胡萝卜素可促进肺成纤维细胞凋亡,增加caspase-12蛋白表达。Objective To explore the effects of thapsigargin on the cell cycle,apoptosis and cysteinyl aspartate specific proteinase（ Caspase-12） protein expression in mouse lung fibroblast stimulated transforming growth factor-β1 （ TGF-β1 ）. Methods Mice lung fibroblast lines frozen in liquid nitrogen were recovered and cultured. Experiment was divided into three groups： the control group,model group and experimental group. RPMI-1640 medium was only add in the control group,5 ng·m L^-1TGF-β1 was add in the model group; while in experimental group, 5 ng · m L^-1TGF-β1 was add for 24 hours, then4 μmol·L^-1thapsigargin was add. After 24 h,the cell cycle and apoptosis were detected by flow cytometry instrument. The protein expression of caspase-12 was examined using the immunohistochemical method.Results In control group,model group and experimental group,theproportion of G1 phase cells were（ 58. 72 ± 8. 35） %,（ 52. 13 ± 7. 61） % and（ 73. 02 ± 9. 32） %,which of S phase cells were（ 29. 50 ± 4. 24） %,（ 40. 54 ± 6. 31） % and（ 21. 66 ± 4. 17） %,G2 phase cells was（ 11. 75 ± 0. 96） %,（ 7. 62 ± 3. 22） % and（ 6. 09 ± 2. 11） % respectively,the differences had statistical significance in G1 phase,S phase and G2 phase among groups（ all P〈0. 05）,while there was no significant difference in G2 phase between experimental group and model group（ P〈0. 05）. The apoptosis rate in control group,model group and experimenta group were 6. 73 %, 5. 03 % and 21. 53 % respectively, the protein expression of Caspase-12 were3356. 45 ± 205. 74,2645. 56 ± 168. 36 and 5424. 76 ± 241. 79,the differences in above two indicators were statistical significant（ all P〈0. 05）. Conclusion Thapsigargin could promote the apoptosis of lung fibroblast and increase the protein expression of caspase-12.

关 键 词：肺成纤维细胞 毒胡萝卜素 细胞周期 凋亡 半胱氨酸特异性蛋白酶-12 

分 类 号：R97[医药卫生—药品]