Effect of pills for replenishi Qi and securing the external on the expression of SIRT5 in mitochondria of COPD cell models
Objective It is to investigate the effect of pills for replenishi Qi and securing the external(PRQSE)on the expression of SIRT5 in mitochondria of bronchial epithelioid cells and its regulatory effect on mitochondrial function in chro-nic obstructive pulmonary disease(COPD).Methods The human bronchial epithelial-like cells 16HBE were stimulate by cigarette smoke extract(CSE)to establish COPD models.6 groups were set up in the experiment:the normal group in which the human bronchial epithelial-like cells 16HBE were normal cultured,the COPD group in which the modeled cells were normal cultured,the COPD + siRNA in which the modeled cells were cultured after siRNA sham-transfection,the COPD +siRNA SIRT5 group in which the modeled cells were cultured after siRNA transfection,the COPD +PRQSE group in which the modeled cells were cultured with medicated serum containing PRQSE,the COPD + siRNA SIRT5 + PRQSE group in which the modeled cells were cultured with medicated serum containing PRQSE after siRNA transfection.The best SIRT5 knockdown sequence was screened by siRNA transfection and RT-qPCR,the changes of cell morphology and mito-chondrial membrane potential in each group were observed under the microscope,the mitochondrial membrane potential and reactive oxygen species(ROS)content of cells in each group were detected by flow cytometry,the ATP content of cells in each group was determined by ELISA,and the relative expression of SIRT5 mRNA in cells of each group was detected by RT-qPCR.Results Compared with the normal group,the cells in each group of COPD were wrinkled and rounded,with low cell density and poor cellular status;the mitochondrial membrane potential,ATP content and relative expression of SIRT5 mRNA were significantly lower(all P＜0.05),and the ROS content was significantly higher(P＜0.05).Compared with the COPD group,the cellular status was worse in the COPD +siRNA SIRT5 group,and the mitochondrial membrane poten-tial,ATP content and relative expression of SIRT5 mRNA were significantly lower(all P＜0.05),and the ROS content was significantly higher(P＜0.05);compared with the COPD group and COPD +siRNA SIRT5 group,the cellular status was significantly improved in the COPD + PRQSE group and COPD + siRNA SIRT5 + PRQSE group,the mitochondrial membrane potential,ATP content and relative expression of SIRT5 mRNA were all significantly higher(all P＜0.05),and the ROS content was significantly lower(P＜0.05).Conclusion Pills for replenishi Qi and securing the external can im-prove mitochondrial dysfunction via regulating the expression of SIRT5 in mitochondria of 16HBE cells induced by CES,thus to play a role in the treatment of COPD.
chronic obstructive pulmonary diseasepills for replenishi Qi and securing the externalmitochondriaSIRT5
NETL
NSTL
万方数据
维普
