Long-term Effect Of Tiao-bu Fei-shen Granules On Rats’ Airway Remodeling With Chronic Obstructive Pulmonary Disease Through Regulate NFκB/TGF-β1/Smad2Signaling

ObjectiveTo evaluate the mechanisms of the long-term effect of three Tiaobu Feishen (BufeiJianpi, Bufei Yishen, Yiqi Zishen) granules of traditional Chinese medicine (TCM) on rats’airway remodeling with chronic obstructive pulmonary disease (COPD), and provideexperimental evidences for practice in clinical.MethodsSprague-Dawley rats were randomized into control, model, Bufei Jianpi, Bufei Yishen,Yiqi Zishen and aminophyline groups. COPD rat model was prepared by smoke inhalationsand repeated bacterial infections. At the period from week9to week20, control and modelgroups was intragastriclly administrated with normal saline, while Bufei Jianpi group withBufei Jianpi granule, Bufei Yishen group with Bufei Yishen granule, Yiqi Zishen group withYiqi Zishen granule and aminophylline group with aminophyline. At week20, half of theanimals were sacrificed, while the survivals raised under normal conditions up to the32ndweek, and then sacrificed. The lung function (tidal Volume, VT; Peak Expiratory Flow, PEF;50%Tidal Volume Expiratory Flow, EF50) was measured. Lung morphology was detected.The levels of interleukin (IL)-1β, IL-6, IL-8and tumor necrosis factor (TNF)-α weredetected by immunohistochemistry methods. Expression of phosphorylated nuclear factor-κB(p-NFκB), phosphorylated inhibitory factor-κBα (p-IκBα) proteins were detected bywestern blotting. Expression of TGF-β1mRNA, Smad2mRNA were detected by fluorescencequantitative PCR.Results1General statusSignificant symptoms appeared in model group, such as emaciation, poor spirit, reducedactivities, decreased food and water intake, loose stool, etc. While symptoms mentionedabove alleviated dramatically in three TCM-treated groups and aminophylline group,especially Bufei Jianpi and Bufei Yishen group. Body weight in three TCM-treated groups and Aminophylline increased quickly from the12thweek compared with model group, butthere was no significant statistical difference among the groups (P>0.05).2Lung functionThe levels of VT, PEF, EF50of Model group were significantly lower than Controlgroup from the4thweek (P<0.05, P<0.01). The levels of PEF and EF50in threeTCM-treated groups were significantly higher than model group from the16thweek (P<0.05,P<0.01), and so did VTfrom the20thweek. PEF of Aminophylline group was significantlyhigher only at the20thweek and the28thweek (P<0.05). During the period of the32weeks,there was no difference in the levels of VT, PEF, EF50between three TCM-treated groupsand Aminophylline group(P>0.05).PEF in three TCM-treated groups was higher at the32ndweek compared with that in the20thweek (P<0.05)，PEF of Aminophylline group wasgenerally increasing (P>0.05).3Lung pathologyAt the end of the20thweek, histology of lung impaired obviously in Model group whilealleviated in different degrees in the three TCM-treated groups and Aminophylline group,especially the three TCM-treated groups. At the end of the32ndweek, the three TCM-treatedgroups can improve the pathological state better than Aminophylline group.4Expression of cytokines of lung and airwayAt both week20and32, The expression of IL-1β, IL-8andTNF-α in the model groupwere significantly higher than that in the control group(P<0.01) and concurrently higher thanthe three TCM-treated groups and aminophylline group(P<0.01).The expression of themarkers mentioned above in the three TCM-treated groups were lower than Aminophylline(P<0.01), especially Bufei Jianpi and Bufei Yishen groups(P<0.01). At week32, Theexpression of TNF-α, IL-1β and IL-8of TCM-treated groups were lower than that at week20(P<0.05, P<0.01), while there was no significant statistical difference in aminophyllinegroup (P>0.05).5Expression of p-NFκB and p-IκBα proteins of lung and airwayAt both week20and32, the p-NFκB and p-IκBα proteins in model group weresignificantly higher than that in control (P<0.01), and concurrently higher than that inTCM-treated groups and aminophylline group(P<0.01). In addition, the expression of theproteins mentioned above in three TCM-treated groups were lower than aminophylline(P<0.01), especially Bufei Jianpi and Bufei Yishen group(P<0.01). At week32, Theexpression of p-NFκB and p-IκBα proteins in the three TCM-treated groups were lower thanthat at week20(P<0.05, P<0.01), while there was no significant statistical difference in aminophylline group (P>0.05).6Expression of TGF-β1and Smad2mRNAAt both week20and32, the expression of TGF-β1and Smad2mRNA in model groupwere significantly higher than that in control (P<0.01), and higher than that in TCM-treatedgroups and aminophylline group simultaneously (P<0.01). Further, the expression of theproteins in three TCM-treated groups were lower than aminophylline (P<0.01), especiallyBufei Jianpi and Bufei Yishen group (P<0.01). At week32, the expression of TGF-β1andSmad2mRNA were higher in model group compared with that at week20(P<0.05, P<0.01),conversely, lower in three TCM-treated groups (P<0.01, P<0.05).7Correlation analysisAt week20and32, there are significant positive correlations between the thickness ofairway wall and levels of IL-1β, IL-8, TNF-α, p-NFκB proteins, p-IκBα proteins,TGF-β1mRNA and Smad2mRNA that contained in the tissue of lung and airway (week20:r=0.974,0.945,0.964,0.954,0.948,0.948,0.964; week32: r=0.977,0.974,0.980,0.971,0.963,0.954,0.968); there are significant negative correlations between the levels of VToflung function and the levels of IL-1β, IL-8, TNF-α, p-NFκB proteins, TGF-β1mRNA andSmad2mRNA that contained in the tissue of lung and airway (week20: r=-0.579,-0.646、-0.652,-0.598,-0.604,-0.622; week32: r=-0.514,-0.555,-0.527,-0.474,-0.509,-0.551).there are significant negative correlations between the levels of PEF of lung function and thelevels of IL-1β, IL-8, TNF-α, p-NFκB proteins, TGF-β1mRNA and Smad2mRNA thatcontained in the tissue of lung and airway (week20: r=-0.487,-0.538,-0.512,-0.439,-0.378,-0.482; week32: r=-0.504,-0.536,-0.526,-0.499,-0.498,-0.551); there are significantnegative correlations between the levels of EF50of lung function and the levels of IL-1β,IL-8, TNF-α, p-NFκB proteins, TGF-β1mRNA and Smad2mRNA that contained in thetissue of lung and airway (week20: r=-0.523,-0.638,-0.594,-0.566,-0.529,-0.589; week32:r=-0.511,-0.548,-0.520,-0.510,-0.496,-0.548).Conclusion1Bufei Jianpi granule, Bufei Yishen granule and Yiqi Zishen granule can obviouslyimprove the general condition, pulmonary function, ease mophological changes in ratswith COPD in the stable period than aminophuline, and have good long-term effects.2Bufei Jianpi granule, Bufei Yishen granule and Yiqi Zishen granule can decrease theexpression of inflammation cytokines, reduce inflammation and improve airwayremodeling through regulate NFκB/TGF-β1/Smad2signaling, and have good long-termeffect. 3The effect of Bufei Jianpi granule and Bufei Yishen granule are better than Yiqi Zishengranule.