Mechanism study of Huashi Baidu Powder in improving pulmonary fibrosis by reducing collagen cross-linking
Objective:To explore therapeutic effect and potential mechanism of Huashi Baidu Powder(HSBD)on pulmonary fibrosis in mice.Methods:Fifty C57BL/6J mice were randomly divided into five groups:the normal group,sham surgery group,bleomycin group,HSBD group(early intervention),and HSBD group(late intervention).A model of idiopathic pulmonary fibrosis(IPF)in mice was established by tracheal injection of bleomycin.RNA-Seq was used to screen for similarities and differences in gene expression levels among the groups,predicting their involvement in biological processes and pathways.Mouse weight changes were recorded,lung function(respiratory rate and flow)was measured,lung CT images were taken,lung tissue collagen morphology was observed using two-photon fluorescence microscopy,tissue pathological changes were assessed using HE staining,and qPCR was employed to detect mRNA levels of hypoxia-inducible factor 1α(HIF1α),lysyl oxidase(LOX),lysyl oxidase-like subtypes(LOXL)1-LOXL4,and lysyl hydroxylase 2(PLOD2)in lung tissue.Immunohistochemical staining was conducted to evaluate the protein expression levels of HIF1α,LOXL2,LOXL3,and PLOD2 in lung tissue.Results:Transcriptome results indicate that HSBP primarily affects collagen synthesis,extracellular matrix interactions,and inflammatory responses,with its mechanism potentially involving the HIF-1 pathway.Compared to the late intervention group,the early intervention group of HSBP accelerates weight recovery,alleviates lung collagen crosslinking,slows down the fibrotic process,and has a positive regulatory effect on the recovery of respiratory rate and flow.Simultaneously,it can reduce the gene expression levels of HIF1α,LOX,and LOXL3 in mouse lung tissue(P＜0.05,P＜0.01),also downregulate the protein expression levels of HIF1α,LOXL2,LOXL3,and PLOD2(P＜0.01).Conclusion:HSBP can reduce collagen cross-linking levels in pulmonary fibrosis,slowing down the fibrotic process.
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