Therapeutic mechanism of emodin for treatment of rheumatoid arthritis: a network pharmacology-based analysis / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic mechanism of emodin in the treatment of rheumatoid arthritis (RA) using a network pharmacology-based method and validate this mechanism in a fibroblast-like synovial cell line.@*METHODS@#The PubChem, Targetnet, SwissTargetPrediction, Genecards, OMIM, and DisGeNET databases were searched to obtain emodin targets and RA-related genes. A protein-protein interaction (PPI) network was constructed, and GO and KEGG pathway enrichment analyses were carried out to analyze the intersection genes. AutoDock4.2.6 software was used to simulate molecular docking between emodin and its candidate targets. In a cultured fibroblast-like synovial cell line (MH7A), the effects of different concentrations of emodin on proliferation of tumor necrosis factor-α (TNF-α)-induced cells were investigated using CCK-8 assay, cell scratch experiment and flow cytometry; the changes in the expressions of nuclear factor-κB (NF-κB) pathway proteins were detected using Western blotting, and the mRNA expressions of the hub genes were examined with RT-qPCR.@*RESULTS@#We identified 32 intersection genes of emodin and RA, and the key targets including CAPS3, ESR1, and MAPK14 involved mainly the NF-κB signaling pathway. Cell scratch experiment and flow cytometry demonstrated a strong inhibitory effect of emodin on MH7A cell proliferation. Treatment with TNF-α significantly increased the cellular expressions of the NF-κB pathway proteins, which were obviously lowered by treatment with 80 μmol/L emodin. The results of RT-qPCR showed that TNF-α treatment obviously up-regulated the expressions of the hub genes COX2 and P38MAPK, and emodin treatment significantly down-regulated the expressions of MAPK and PTGS2 and up-regulated the expression of CASP3.@*CONCLUSION@#The therapeutic effect of emodin on RA is mediated mainly through regulation of cell proliferation, apoptosis, and the NF-κB pathway.

Effect of Electroacupuncture on Inflammatory Response of Rats after Focal Cerebral Ischemia-reperfusion via Regulating IκB Kinases β Expression / 中国康复理论与实践

Objective:To observe the changes of inflammatory damage in the brain of rats after focal cerebral ischemia-reperfusion, and explore the effect of the initiation of IκB kinases β (IKKβ), which is the key protein of activating nuclear factor (NF)-kappa B signaling pathway in inflammatory response, and the mechanism of electroacupuncture inhibiting inflammatory damage. Methods:A total of 240 male Sprague-Dawley rats were randomly divided into sham group, ischemia-reperfusion group, electroacupuncture group, IKKβ silencing group, IKKβ overexpression group and IKKβ overexpression + electroacupuncture group, each group was further divided into six hours, twelve hours, 24 hours, 48 hours and 72 hours subgroups. The right middle cerebral artery occlusion reperfusion model was established by modified thread embolization. The IKKβ gene was intervened by gene silencing and gene overexpression technology. Results:Compared with the model group, the neurological function score increased (P < 0.05), the cerebral infarction volume decreased (P < 0.05), the activation of NF-κB p65 was inhibited, and the content of proinflammatory factors decreased (P < 0.05) in IKKβ silencing group. Compared with IKKβ silencing group, the above results were significantly worse in IKKβ overexpression group (P < 0.05), and microglia in cerebral ischemic cortex were significantly activated. The activation of microglia and activation of IKKβ were significantly inhibited in IKKβ overexpression + electroacupuncture group. Conclusion:IKKβ gene silencing could inhibit the inflammatory response of cerebral ischemic cortex mediated by NF-κB signaling pathway, and over-expression of IKKβ could lead to severe inflammatory damage in ischemic cortex. Electroacupuncture could inhibit the inflammatory damage after focal cerebral ischemia-reperfusion by regulating the activity of IKKβ.