基于网络药理学探讨甘露消毒汤干预转基因小鼠SARS-CoV-2假病毒感染后嗅觉障碍的机制OA

Objective To explore the mechanism of Ganlu Xiaodu Decoction on olfactory dysfunction in hACE2 transgenic mice infected with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)postviral infection olfactory disturbance(POVD)based on network pharmacology and molecular docking.Methods Active components and action targets of Ganlu Xiaodu Decoction were collected from TCMSP database.The olfactory dysfunction targets were obtained through GeneCards database,and the intersection of drugs and disease targets was obtained;the drug-active components-targets-disease network was constructed by using Cytoscape 3.8.0 software,and the protein interaction network was constructed by using STRING database.The GO and KEGG pathways of key targets were enriched and analyzed.AutoDock Vina and PyMOL software were used to conduct molecular docking between the main active components and the core target.The hACE2 transgenic mice were randomly divided into normal group,model group and Ganlu Xiaodu Decoction low-,medium-and high-dosage groups.Except the normal group,all mice were given 1×109/mL SARS-CoV-2 pseudovirus venom nasal drops(0.05 mL/mouse).The olfactory function of mice was evaluated by buried food pellet test(BFPT)after intervention for 7 days.HE staining was used to observe the histomorphology of olfactory mucosa in mice.ELISA was used to detect the contents of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-10,interferon-γ(IFN-γ)and C-C motif chemokine ligand 5(CCL5)in serum.Immunohistochemical staining was used to detect the expressions of TNF-α,IL-6 in olfactory mucosa and olfactory marker protein(OMP)in olfactory bulb.Results Totally 63 active components of Ganlu Xiaodu Decoction were identified,along with 235 targets.Among these,20 potential target molecules were identified that may intervene in POVD,primarily including AKT1,TNF and IL-6,etc.The key pathways were human cytomegalovirus infection signaling pathway,neurodegenerative disease multiple signaling pathway,influenza a disease signaling pathway,etc;molecular docking results showed that baicalin,kaempferol,quercetin,aloe emodin and luteolin exhibited high docking affinity with core targets AKT1,TNF and IL-6,indicating significant binding advantages between key bioactive components and core targets.Animal experiments indicated impaired olfactory function in mice assessed by BFPT.In Ganlu Xiaodu Decoction medium-and high-dosage groups,the number of inflammatory cells in the olfactory mucosa of mice decreased significantly;swelling was reduced,and mucosal thickness gradually returned to normal,the cellular structure and morphology of olfactory sensory neurons tended toward regularity and integrity;serum contents of TNF-α,IL-6,IFN-γ and CCL5 decreased,while IL-10 content increased(P<0.05,P<0.01);the expressions of TNF-α and IL-6 in the olfactory mucosa decreased(P<0.01,P<0.001),with upregulation trend of OMP expression in the olfactory bulb(P>0.05).Conclusion Ganlu Xiaodu Decoction acts on AKT1,TNF,IL-6 and other targets through wogonin,kaempferol,quercetin,aloe emodin,luteolin and other components,down-regulates the expressions of a variety of cytokines,reduces inflammatory injury of olfactory mucosa,and improves the expression of OMP in olfactory bulb,so as to treat POVD.