摘要
目的探究欧前胡素衍生物(IMP-1)对慢性阻塞性肺疾病(COPD)肺泡Ⅱ(ATⅡ)型细胞活性及耐药蛋白的影响。方法ATⅡ型细胞分为空白组(常规培养基进行培养的生长良好的ATⅡ型细胞)、香烟烟雾提取物(CSE)组(加入2%CSE诱导培养)、IMP-1组(采用10μmol/L的IMP-1干预经CSE诱导的ATⅡ型细胞)。采用四甲基偶氮唑盐(MTT)法测定ATⅡ型细胞24 h、48 h、72 h增殖率,Hoechst 33342染色法检测ATⅡ型细胞凋亡率,流式细胞分析仪分析各周期细胞占比,软琼脂克隆形成实验检测ATⅡ型细胞克隆数量,Western blot法检测多耐药基因1(MDR1)及多药耐药相关蛋白1(MRP1)蛋白表达水平。结果与空白组比较,CSE组细胞凋亡率、ATⅡ型细胞周期G0/G_(1)期占比升高,在24 h、48 h、72 h细胞的增殖率和S、G_(2)/M期占比降低(P<0.05);与CSE组比较,IMP-1组细胞增殖率和S、G_(2)/M期占比及细胞克隆数量升高,ATⅡ型细胞周期G0/G_(1)期占比和MDR1、MRP1蛋白降低(P<0.05)。结论IMP-1可能通过下调CSE诱导的ATⅡ型细胞的MDR1、MRP1蛋白表达,从而抑制细胞的异常活化,增加其凋亡。
Objective To explore effects of a derivative of imperatorin(IMP-1)on the activity and resistance proteins of alveolar typeⅡ(ATⅡ)cells in chronic obstructive pulmonary disease(COPD).Methods ATⅡcells were divided into the blank group(well growing ATⅡcells cultured in conventional medium),the cigarette smoke extract(CSE)group(induced culture with 2%CSE added)and the IMP-1 group(intervention with 10μmol/L of IMP-1 on CSE induced ATⅡcells).Proliferation rates of ATⅡcells at 24,48 and 72 hours were measured using MTT method.Apoptosis of ATⅡcells was detected by Hoechst 33342 staining.The proportion of cells in each cycle was analyzed by flow cytometry.The number of ATⅡcell clones was detected by soft AGAR clone formation assay.Multidrug resistance gene 1(MDR1)and multidrug resistance associated protein 1(MRP1)were detected by Western blot assay.Results Compared with the blank group,the apoptosis rate and the proportion of G0/G_(1) phase of ATⅡcell cycle were increased in the CSE group,and the proliferation rate and the proportion of S,G_(2)/M phase were decreased at 24 h,48 h and 72 h(P<0.05).Compared with the CSE group,the cell proliferation rate,the proportion of S and G_(2)/M phases and the number of cell clones were increased in the IMP-1 group,and the proportion of G0/G_(1) phase in ATⅡcell cycle and the protein levels of MDR1 and MRP1 were decreased in the IMP-1 group(P<0.05).Conclusion IMP-1 may downregulate the expression of MDR1 and MRP1 proteins induced by CSE in ATⅡcells,inhibiting the progression of ATⅡcells from S phase to G_(2) phase and from G_(2) phase to M phase,thereby inhibiting abnormal cell activation and increasing apoptosis.
作者
于志鸿
王小琴
YU Zhihong;WANG Xiaoqin(Department of Respiration,Urumqi Fifth People's Hospital,Urumqi 830013,China;Department of Geriatrics,Urumqi Fifth People's Hospital,Urumqi 830013,China)
出处
《天津医药》
CAS
2024年第11期1127-1131,共5页
Tianjin Medical Journal
关键词
肺疾病
慢性阻塞性
欧前胡素衍生物
肺泡Ⅱ型上皮细胞
香烟烟雾提取物
多耐药基因1
多药耐药相关蛋白1
pulmonary disease
chronic obstructive
imperatorin derivatives
alveolar typeⅡepithelial cells
cigarette smoke extract
multidrug resistance gene 1
multidrug resistance associated protein 1
