Effect of Zijin Bushui Ointment on rats with chronic obstructive pulmonary disease based on the miR-30b-5p/BCL2L11 axis and the underlying mechanism
Objective To explore the effect of Zijin Bushui Ointment on rats with chronic obstructive pulmonary disease(COPD)rats and its potential mechanism.Methods Eighty-four male Wistar rats in specific pathogen free(SPF)level were selected and randomly assigned into 7 groups(12 rats/group)according to the random number table method:control group,model group,low-dose,medium-dose and high-dose Zijin Bushui Ointment groups,antagomir NC group(high-dose Zijin Bushui Ointment+transfection of antagomir NC),and miR-30b-5p antagomir group(high-dose Zijin Bushui Ointment+transfection of miR-30b-5p antagomir).Except for rats in the control group,the COPD rat model was constructed by airway instillation of lipopolysaccharide(LPS)combined with smoke exposure.After successful modeling,rats in each group were given corresponding drugs by oral gavage and tail vein injection,once a day for 12 weeks.Small animal lung function testing instrument was used to detect the lung function of rats in each group.The mRNA expression of miR-30b-5p in lung tissue was detected by real-time fluorescence quantitative PCR(RT-qPCR).Hematoxylin and eosin(H&E)staining was performed to visualize pathological damage of lung tissue and measure the thickness of small airway wall in lung tissue.Terminal deoxynucleotidyl transferase dUTP nick-end labeling(TUNEL)staining was applied to observe the apoptosis of rat lung epithelial cells.Serum interleukin(IL)-8 and IL-β levels were detected by enzyme linked immunosorbent assay(ELISA).Western blot was applied to detect the protein expressions of anti-apoptotic factor B cell lymphocyte tumor 2(Bcl-2),Bcl-2 associated X protein(Bax)and BCL2L11.Dual luciferase reporter assay was applied to verify the targeting relationship between miR-30b-5p and BCL2L11.Results Compared with those of the control group,rats in the model group had significantly higher resting respiratory rate,resistance index(RI),thickness of small airway wall in lung tissue,apoptosis index of lung epithelial cells,serum levels of IL-8 and IL-β,and protein expressions of Bax and BCL2L11,severer pathological damages in the lung,and significantly lower peak inspiratory flow(PIF),compliance dynamic(Cdyn),mRNA level of miR-30b-5p,and the protein expression of Bcl-2(P<0.05).Compared with those of Model group,rats in the low-dose,medium-dose and high-dose Zijin Bushui Ointment groups had significantly lower resting respiratory rate,RI,thickness of small airway wall in lung tissue,apoptosis index of lung epithelial cells,serum levels of IL-8 and IL-β,and protein expressions of Bax and BCL2L11,milder pathological damages in the lung,and significantly higher PIF,Cdyn,mRNA level of miR-30b-5p,and the protein expression of Bcl-2(P<0.05).There was no significant difference in resting respiratory rate between the medium-dose and low-dose Zijin Bushui Ointment groups(P>0.05),and significant differences in the other indicators were detected in the pairwise comparison among the low-dose,medium-dose and high-dose Zijin Bushui Ointment groups(P<0.05).Compared with those of high-dose Zijin Bushui Ointment group and antagomir NC group,rats in the miR-30b-5p antagomir group had significantly higher resting respiratory rate,RI,thickness of small airway wall in lung tissue,apoptosis index of lung epithelial cells,serum levels of IL-8 and IL-β,and protein expressions of Bax and BCL2L11,severer pathological damages in the lung,and significantly lower PIF,Cdyn,mRNA level of miR-30b-5p,and the protein expression of Bcl-2(P<0.05).Transfection of miR-30b-5p antagomir weakened the protective effect of high-dose Zijin Bushui Ointment on COPD.MiR-30b-5p negatively regulated the expression of BCL2L11.Conclusion Zijin Bushui Ointment down-regulates BCL2L11 by regulating miR-30b-5p,thereby inhibiting inflammatory response,reducing apoptosis of lung epithelial cells,improving lung function,and alleviating symptoms of COPD rats.
万方数据
维普
