Exploring Regulatory Mechanism of Ganlu Xiaodu Decoction(甘露消毒汤)on SARS-CoV-2 Pseudovirus Infected hACE2 Transgenic Mice Based on Signal Pathway TGF-β1/Smad3
Objective Using SARS-CoV-2 pseudovirus technology,the expression changes of TGF-β1/Smad3 signaling pathway related proteins were investigated to explore the mechanism of Ganlu Xiaodu Decoction(甘露消毒汤)on SARS-CoV-2 infection.Methods Sixty experimental mice were randomly assigned to three groups:normal group,model group,Ganlu Xiaodu Decoction group.Except the normal group,the mice in the other two groups were given nasal drops of SARS-CoV-2 pseudovir-us for modeling.Following modeling,the drug intervention was administered,and the samples were collected on the seventh day of treatment.The expressions of interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),interleukin 1 β(IL-1 β)and inter-leukin-6(IL-6)in serum were determined using ELISA method.The mRNA contents of transforming growth factor-β1(TGF-β1),Smad3,CD9 and CD63 in lung tissue were quantified using qPCR method.The protein expressions of TGF-β1,Smad3,CD9 and CD63 were analyzed by Western blot(WB)assay.Results Model group:The lung index and pathological score of lung tissue in mice were increased.After HE staining,HE staining revealed thickening of alveolar septa,infiltration of inflammatory cells and increased expressions of TGF-β1,Smad3,CD9 and CD63 mRNA and protein in lung tissue.Results Serum levels of TNF-α,IL-1 β and IL-6 were elevated,while IL-10 expression was decreased(P<0.05).Ganlu Xiaodu Decoction group:Lung index and lung tissue pathological score were significantly down-regulated(P<0.05).HE staining demonstrated reduced inflammatory cell infiltration and decreased expressions of TGF-β1,Smad3,CD9 and CD63 mRNA and protein in mouse lung tissue(P<0.05).HE staining showed reduced inflammatory cell infiltration,decreased expressions of TGF-β1,Smad3,CD9 and CD63mRNA and protein in mouse lung tissue(P<0.05),decreased serum levels of TNF-α,IL-1 β and IL-6 and in-creased IL-10 content(P<0.05).Conclusion Ganlu Xiaodu Decoction can down-regulate inflammatory factors(IL-6,TNF-α,IL-1 β),up-regulate anti-inflammatory factor(IL-10),inhibit the process of inflammatory pathological reactions and alleviate the inflammatory response of lung tissue in mice with pseudovirus.Its mechanism may be related to the inhibition of pathway TGF-β1/Smad3.
pseudovirusGanlu Xiaodu Decoction(甘露消毒汤)TGF-β1hACE2Spike protein