雷公藤红素调控Notch途径对慢阻肺大鼠肺成纤维细胞气道影响的研究OA

Objective To investigate the effects of celastrol(Cel)on airway remodeling factors,collagen synthesis and degradation in lung fibroblasts of chronic obstructive pulmonary disease(COPD)rats by regulating the neurogenic locus notch homolog protein(Notch)pathway.Methods A total of 30 rats were randomly divided into 3 groups:animal control group(normal feeding),animal model group(COPD rat model established by smoke exposure combined with lipopolysaccharide tracheal instillation)and animal experimental group(intraperitoneal injection of 0.01 mg·kg-1 Cel after successful modeling).The bronchial fibroblasts(BF)cells were isolated from rats in the animal model group and randomly divided into cell control group(no treatment),cell experimental group(2 μmol·L-1 Cel)and inhibitor group[1 mmol·L-1 valproic acid(VPA)].Pulmonary function was assessed in rats using invasive pulmonary function testing methods;Western blot was used to detect the expression of Notch signaling pathway-related proteins in each group of cells;real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect mRNA expression of airway remodeling-related factors;enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of matrixmetalloproteinase-1(MMP-1)and tissue inhibitors of metalloproteinase-1(TIMP-1)in each group of cells.Results The forced vital capacity(FVC)of rats in animal control group,animal model group and animal experimental group were(6.74±0.87),(3.28±0.51)and(4.94±0.83)mL,respectively;the forced expiratory volume in 0.1 second(FEVO.1)were(3.94±0.45),(1.05±0.16)and(2.41±0.37)mL,respectively;the peak expiratory flow(PEF)were(28.09±4.60),(16.63±3.21)and(21.55±4.02)L·min-1,respectively.Significant differences were observed in the above indicators between animal control group and animal model group,and between animal model group and animal experimental group(P＜0.01,P＜0.001).The relative expression levels of(Notch1)protein in cell control group,cell experimental group and inhibitor group were 1.00±0.19,0.12±0.02 and 0.38±0.06,respectively;the relative expression levels of Notch1 receptor intracellular binding domain(NICD1)protein were 1.00±0.17,0.72±0.12 and 0.85±0.13,respectively;the relative expression levels of serrated typical Notch ligand 1(Jagged 1)protein were 1.00±0.15,0.71±0.11 and 0.84±0.12,respectively;the relative expression levels of hair and enhancer of split 1(Hes1)protein were 1.00±0.11,0.69±0.08 and 0.81±0.09,respectively;the relative expression levels of MMP-9 mRNA were 1.00±0.16,0.59±0.08 and 0.73±0.11,respectively;the relative expression levels of type Ⅰ collagen(Col Ⅰ)mRNA were 1.00±0.14,0.67±0.09 and 0.86±0.14,respectively;the relative expression levels of α-smoth muscle actin(α-SMA)mRNA were 1.00±0.16,0.41±0.07 and 0.78±0.13,respectively;the relative expression levels of transforming growth factor-β(TGF-β)mRNA were 1.00±0.18,0.53±0.09 and 0.75±0.14,respectively;the contents of MMP-1 were(1.19±0.17),(0.80±0.15)and(1.06±0.15)ng·mL-1,respectively;the contents of tissue inhibitor of protease-1(TIMP-1)were(219.78±30.24),(131.94±18.15)and(167.51±21.01)ng·mL-1,respectively.There were all statistically significant differences in the above indicators between cell control group and cell experimental group,as well as between cell experimental group and inhibitor group(P＜0.05,P＜0.01,P＜0.001).Conclusion Celastrol can effectively improve lung function and pulmonary pathological damage in COPD rats,reduce the levels of inflammatory cells and inflammatory factors,and improve the imbalance of airway remodeling-related factors and collagen synthesis/degradation in BF cells,which may be related to its inhibition of Notch signaling activation.