The Mechanism of Jiawei Jinkui Shenqi Pills Improving Asthma Rats with Kidney Yang Deficiency Syndrome by Interfering with Ferroptosis through MUC5AC/EGFR Pathway
[Objective]To investigate the possible mechanism of action of Jiawei Jinkui Shenqi Pill in relieving asthma with kidney Yang deficiency.[Methods]Total 40 SD rats were randomly divided into control group,model group,Jiawei Jinkui Shenqi Pill group(referred to as Chinese medicine group),and dexamethasone group(referred to as DXM group),10 rats in each group,and all other groups except control group were induced by the combination of hydrocortisone injection and ovalbumin(OVA) to construct the rat model of kidney Yang deficiency asthma.The body mass of the rats was monitored regularly and wheezing symptoms were evaluated.Hematoxylin-eosin(HE) and periodic acid-Schiff(PAS) staining were used to observe the pathological morphology of rat lung tissues and bronchi;enzyme-linked immunosorbent assay(ELISA) was used to detect relevant indices in rat serum;and Western blot was used to detect protein levels of Mucin 5AC(MUC5AC),epidermal growth factor receptor(EGFR),and glutathione peroxidase 4(GPX4);and Real-time quantitative polymerase chain reaction(RT-qPCR) to detect the expression of MUC5AC,EGFR,GPX4 mRNA.Human bronchial epithelial cells BEAS-2B were cultured and divided into blank control group,ferric amine citrate(FAC) group,MUC5AC group,FAC+MUC5AC group.The concentration of non-haem iron was detected with red feldspun microplate method;the level of reactive oxygen species(ROS) was observed with fluorescence staining,and comparative analysis of the correlation between the MUC5AC pathway and ferroptosis.[Results]Compared with control group,the lung tissue bronchial tube wall thickened,a large number of inflammatory cells infiltrated,there was goblet cell hyperplasia in rats of model group;serum levels of interleukin-4(IL-4),interleukin-5(IL-5),interleukin-13(IL-13) significantly increased (P<0.05),cortisol(COR) levels were significantly decreased(P<0.05),MUC5AC and EGFR mRNA expression was increased,GPX4 mRNA expression was decreased(P<0.05),MUC5AC and EGFR protein content was increased and GPX4 protein content was decreased(P<0.05).Compared with model group,serum levels of IL-4,IL-5,IL-13 decreased and COR increased in Chinese medicine group and DXM group(P<0.05);MUC5AC,EGFR levels decreased and GPX4 protein level increased(P<0.05);MUC5AC and EGFR mRNA down-regulated and GPX4 mRNA expression up-regulated(P<0.05).Compared with blank control group,non-heme iron concentration significantly increased in the other groups(P<0.05),the level of ROS increased,and the FAC+MUC5AC group is more pronounced than the MUC5AC,FAC medication group.[Conclusion]Jiawei Jinkui Shenqi Pill can intervene ferroptosis and improve asthma of kidney yang deficiency syndrome by regulating MUC5AC/EGFR pathway.The mechanism may be to reduce the excessive secretion of mucus and reduce the absorption rate of iron ions by inhibiting the overexpression of MUC5AC,and reduce ferroptosis.
Jiawei Jinkui Shenqi PillMUC5ACEGFRGPX4ferroptosiskidney Yang deficiencybronchial asthmamechanism research
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