Study on mechanism of Chenpi in treatment of anemia of chronic disease with Yigong Powder based on network pharmacology and experimental validation
Objective To investigate the mechanism by which Chenpi(Citri Reticulatae Pericarpium)contributes to Yigong Powder's treatment of anemia of chronic disease(ACD),this study analyzes the different targets and signaling pathways of Chenpi and other active components of Sijunzi Tang in Yigong Powder using network pharmacology,and validates the role of Chenpi's active compounds in ACD treatment through animal experiments.Methods ①Network pharmacology:Active components of Yigong Powder were obtained from the Traditional Chinese Medicine Systems Pharmacology Database(TCMSP),HERB,and HIT databases,based on oral bioavailability(OB)≥30%and drug-likeness(DL)≥0.18,or by screening for absorption,distribution,metabolism,and excretion standards through the SwissADME database.The targets of the active components were obtained through the TCMSP and the SwissTargetPrediction database.The disease targets ACD were then identified using the human gene database(GeneCards),the online mendelian inheritance in man(OMIM)database,and the disease-centric interaction database among diseases and various associated genes(DisGeNET).Intersection targets were identified using the Venny 2.1 platform.Protein interaction networks were constructed using the STRING database and Cytoscape v3.9.1 software.The top 30 core targets were screened using the MCC algorithm,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis of the core targets was performed.Cytoscape was used to construct a"Yigong Powder active component-ACD-target"network and a"core target-signaling pathway"network.②Molecular docking validation:The distinct targets of Chenpi,compared to other active components in Yigong Powder,were validated through molecular docking analysis.③An ACD mouse model was prepared and treated with drug interventions.After one week of intervention,the levels of white blood cells(WBC),hemoglobin(Hb),red blood cells(RBC),serum ferritin(SF),and serum iron(SI)were measured and compared across groups.Histopathological changes in spleen tissue were observed using hematoxylin and eosin(HE)staining.Western blot analysis was performed to detect the expression of phosphorylated MEK(p-MEK)/MEK,phosphorylated extracellular signal-regulated kinases 1/2(p-ERK1/2)/ERK1/2,and phosphorylated P65(p-P65)/P65 in spleen tissue,to verify the therapeutic effects and mechanisms of Chenpi's active components in the treatment of ACD.Results ①Network pharmacology analysis revealed 135 active components in Yigong Powder,acting on 1,006 targets.There were 1,474 disease targets of ACD,with 235 common targets between the active components of Yigong Powder and ACD.The top 30 core targets were identified and used to construct the"Yigong Powder active component-ACD-core target"network and the"core target-signaling pathway"network.It was found that the active component of Chenpi,naringenin,and its target,mitogen-activated protein kinase 3(MAPK3),may play a crucial role in the treatment of ACD through the MEK-ERK1/2-NF-κB signaling pathway.②Animal experiment results showed that,compared to the mice in model group,the naringenin-treated group mice showed significantly lower levels of WBC and SF(P<0.05),and significantly higher levels of Hb,RBC,and SI(P<0.05)in peripheral blood.Western blot analysis showed a significant decrease in the expression of p-MEK/MEK,p-ERK(1/2)/ERK(1/2),and p-P65/P65 proteins in spleen tissue(P<0.05).HE staining indicated a significant reduction in necrotic areas and neutrophil infiltration in spleen tissue.Conclusion In Yigong Powder,the active component of Chenpi,naringenin,reduces the activation of MAPK3 protein to downregulate the MEK-ERK1/2-NF-κB pathway,leading to a reduction in inflammation in ACD mice,correction of RBC homeostasis,regulation of iron metabolism,and improvement of anemia.
anemia of chronic diseaseYigong PowderCitri Reticulatae Pericarpiumnaringeninnetwork pharmacologytraditional Chinese herbal medicine research
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