Pingchuan Formula Improves Airway Remodeling by Upregulating miR-23b Expression in BALF of Asthmatic Mice
To investigate the mechanism of Pingchuan formula'intervention in TGF-β/EMT by upregulating exosome miR-23b and its improvement in OVA-induced airway remodeling in asthmatic mice,40 BALB/c male mice were divided into blank group,asthma model group,Pingchuan fumula group,and dexamethasone group according to the random number table,with 10 mice in each group.The 1 d and 14 d model group,the Pinchuan formula group,and the dexamethasone group were sensitized by intraperitoneal injection of 10％OVA+aluminum-magnesium adjuvant,stimulated by atomization of 2.5％OVA every other day from the 21 d to 49 d,and were given daily intragastric administration for 28 d,and samples were taken 24 h after the last administration.Blank group and model group were given the same amount of normal saline intragastric administration,and samples were taken 24 h after the last administration.Hematoxylin-eosin(HE)and Masson staining were used to observe the lung histopathological changes of mice in each group.The morphology of exosomes in alveolar lavage fluid(BALF)was observed under transmission electron microscopy,and the contents of exosome surface marker proteins CD9,CD63 and CD81 were detected by Western blot.Real-Time fluorescence quan-titative polymerase chain reaction(Real-Time PCR)was used to detect miR-23b mRNA levels in BALF exosomes.The contents of transforming growth factor β1(TGF-β1),smooth muscle actin(α-SMA),Vimentin,E-cadherin and compact connectin 1(ZO-1)were detected by Western blot.CD9,CD63 and CD81 proteins were expressed in the exosomes extracted from BALF of mice in all groups,which proved that the exosomes were successfully extracted in this study.Compared with the blank group,pathological stai-ning showed that the model group of mice had inflammatory cell infiltration around the bronchus,thickened muscle layer of airway wall,and increased collagen fibers.The mRNA level of miR-23b in exosomes was significantly decreased(P＜0.01).The protein levels of Collagen Ⅰ,Collagen Ⅱ,TGF-β1,α-SMA and Vimentin in lung tissues were significantly increased(P＜0.01),while the protein levels of E-cadherin and ZO-1 were significantly decreased(P＜0.01).Compared with model group,the pathologic injury of Pingchuan formula group and dexamethasone group was mild,no obvious inflammatory cell infiltration was observed,airway collagen fibers were decreased,and the level of miR-23b mRNA in exosomes was significantly increased(P＜0.01).The protein levels of Collagen Ⅰ,Collagen Ⅱ,TGF-β1,α-SMA and Vimentin were significantly decreased(P＜0.01,P＜0.05),while the protein levels of E-cadherin and ZO-1 were significantly increased(P＜0.01,P＜0.05).Pingchuan formula can improve airway remodeling in asthmatic mice,and its mechanism may be related to up-regulation of exosomal miR-23b expression,thereby inhibiting TGF-β/EMT signal axis and improving airway remodeling.
NETL
NSTL
万方数据
