The Role of Erbin in GTS-21 Regulating Inflammtory Responses in MDP-Stimulated Macrophages

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To explore the role of Erbin protein and nucleotide-binding oligomerization domain 2/receptor-interacting serine/threonine protein kinases (NOD2/RICK) in GTS-21 activating cholinergic anti-inflammatory pathway.


Experiments were randomly divided into four groups: normal control (NC) group, muramyl dipeptide (MDP) group, 10 μg/mL MDP, GTS-21 (GTS) group, 10 μg/mL MDP plus 50 μg/mL GTS-21 (α7 nAChRs agonist), Erbin shRNA interference (sh-Erbin) group: sh-Erbin RNA plus 10 μg/mL MDP and 50 μg/mL GTS-21. We extract specimens at the point of 1, 6, and 24 h after stimulation of MDP in Raw264.7 macrophages.


After stimulation of MDP, the NLR2 mRNA, RICK and Erbin protein, nuclear factor (NF)-κB activity, the pro-inflammatory cytokines tumor necrosis factor (TNF)-α and HMGB1 were significantly increased in MDP group (P <0.05). The expression peak of TNF-α is at 1 h. The peak of HMGB1 is at 24 h. Compared with MDP group, the NLR2 mRNA, RICK, NF-κB, TNF-α, and HMGB1 were significantly decreased, but the Erbin was increased in GTS group (P <0.05). Compared with GTS group, the NLR2 mRNA, RICK, NF-κB, TNF-α, and HMGB1 increase in sh-Erbin group (P <0.05).


GTS-21 could significantly inhibit MDP-induced pro-inflammatory cytokines responses via activating cholinergic anti-inflammatory pathway, and the Erbin might be the key negative regulatory protein in NLR2/RICK signal transduction.

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