Danggui Buxue Decoction Intervening in ROS/TXNIP/NLRP3 Signaling Pathway to Alleviate Lung Inflammatory Injury in Mice with Chronic Intermittent Hypoxia
Objective:To investigate the ameliorative effects and potential mechanisms of Danggui Buxue Decoction(DBD)on in-flammatory lung injury in mice with chronic intermittent hypoxia(CIH).Methods:Twenty-eight male C57BL/6 mice were randomly di-vided into four groups:normal control group(Control group),CIH group,CIH+DBD group,and DBD group.Mice in the CIH and CIH+DBD groups were placed in a hypoxic chamber which was filled with nitrogen to reduce the oxygen concentration from 21%to 9%within 90 seconds,followed by oxygen to gradually increase the oxygen concentration back to 21%within another 90 seconds.Mice in the Control and DBD groups were placed in the chamber but supplied with normal air.This procedure was conducted for 8 hours each day for 35 consecutive days.Mice in the CIH+DBD and DBD groups received intragastric administration of DBD before entering the chamber each day,while mice in the Control and CIH groups received an equal volume of saline intragastrically at the same time.After the animal modeling was completed,the pathological condition of the lungs of mice in each group was examined.qPCR was used to detect the mRNA expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in lung tissue.DHE probes were used to detect reactive oxygen species(ROS)content in lung tissue.Western Blot analysis was performed to assess the expression levels of thioredoxin-interacting protein(TXNIP),nucleotide-binding oligomerization domain-like receptor pro-tein 3(NLRP3),Caspase-1,IL-1β,superoxide dismutase 2(SOD2),and NADPH oxidase 2(NOX2).Immunohistochemical staining was used to observe the expression levels of TXNIP and NLRP3.Results:Compared with the Control group,mice in the CIH group ex-hibited significantly thickened alveolar septa with extensive inflammatory cell infiltration,accompanied by reduced alveolar count and enlarged alveolar spaces.Additionally,there was an increase in ROS content in lung tissue(P＜0.05),elevated protein expression of TXNIP,NLRP3,Caspase-1,and IL-1β(P＜0.05),increased mRNA levels of TNF-α,IL-6,and IL-1β(P＜0.05),decreased protein expression of SOD2(P＜0.05),and increased protein expression of NOX2(P＜0.05).After intervention with DBD,compared with the CIH group,the CIH+DBD group showed improved alveolar septal thickening,reduced inflammatory cell infiltration,decreased ROS content in lung tissue(P＜0.05),and reduced levels of inflammatory cytokines(P＜0.05).Furthermore,there were decreases in pro-tein expression of TXNIP,NLRP3,Caspase-1,IL-1β,and NOX2(P＜0.05)and an increase in SOD2 expression(P＜0.05).Immuno-histochemical staining results revealed that,compared with the Control group,the CIH group had higher expression of TXNIP and NL-RP3 proteins,predominantly located in the alveolar septa and alveolar epithelial cell cytoplasm,which was alleviated after administration of DBD.Conclusion:Danggui Buxue Decoction has a significant ameliorative effect on lung injury in CIH mice,and its mechanism may be achieved by inhibiting the ROS/TXNIP/NLRP3 signaling pathway.
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