Cilostazol Suppresses IL-23 Production in Human Dendritic Cells via an AMPK-Dependent Pathway

Abstract

Background/Aims: Cilostazol has been previously demonstrated to inhibit IL-23 production in human synovial macrophages via a RhoA/ROCK-dependent pathway. However, whether cilostazol affects IL-23 production in human dendritic cells remains largely unknown. The present study was designed to investigate this question and elucidate the possible underlying mechanisms. Methods: Human monocyte-derived dendritic cells (mo-DCs) were pretreated with or without cilostazol and then incubated with zymosan. Enzyme-linked immunosorbent assay (ELISA) and real time PCR analyses were used to measure IL-23 protein expression and RNA levels, respectively, whereas Western blotting was used to measure the expression and phosphorylation level of AMPK. Results: Our results demonstrated that cilostazol suppressed zymosan-induced IL-23 protein production in a concentration dependent manner without affecting dendritic cell viability. In addition, it was found that cilostazol suppressed the expression of the p19 and p40 subunits of IL-23. Moreover, cilostazol mimicked the effect of the AMPK agonist A-769662, as demonstrated by the fact that IL-23 production was also inhibited by A-769662, and the effect of cilostazol on IL-23 production was blocked by the AMPK antagonist Compound C. More importantly, Western blotting demonstrated that cilostazol led to an increased phosphorylation of AMPK. Conclusion: Collectively, our data suggest that cilostazol inhibits the production of IL-23 in human mo-DCs, potentially via the activation of AMPK. This suggests that cilostazol could be an effective anti-inflammatory agent in IL-23- and dendritic cell-related diseases.

Inhibiting AKT signaling pathway with cilostazol and meloxicam synergism for suppressing K562 cells in vitro

“…[ 39 ] Shi et al have observed that cilostazol has an anti‐inflammatory effect via AMPK activation in human dendritic cells. [ 40 ] Also, Lee et al have shown that cilostazol inhibits alcohol‐induced apoptosis through AMPK activation. [ 32 ] Similarly, in our work, cilostazol elevated AMPK phosphorylation, but, it did not affect the effect of meloxicam in combination drug.…”

“…Based on meloxicam and cilostazol IC50 concentrations, different concentrations (20,40,60, and 80 µM) of combination drugs were prepared and stored at −4°C. Next, the MTT assay was performed as described above.…”

“…diphenyltetrazolium bromide (MTT) assay standard protocol. [24] In brief, 6000 cells per well of 96-well plates were seeded and treated with cilostazol and meloxicam at different concentrations (20,40,60, and 80 μM) and the plates were incubated for 24, 48, and 72 h. After incubation, 10 µl of MTT solution (Sigma, 5 mg/ml) was added to each well and incubated for 4 h. Next, 100 µl of pure DMSO was added to each well for dissolving formazan crystals.…”

Zymosan-a Protects the Hematopoietic System from Radiation-Induced Damage by Targeting TLR2 Signaling Pathway

“…Many studies indicated that Zymosan-A can induce NF-κB phosphorylation and nuclear translocation by targeting TLR2[21, 22, 29]. Zymosan-A can also upregulate cytokines, including TNF-α; the IL-1 family; the IL-6, IL-8, and IL-10 family; IL-11; IL-23; the IL-12 family; IL-15; TGF-β; and G-CSF[30-34]. In addition, IL-6, IL-11, IL-12, G-CSF, and TNF-α are regarded as radiation protection factors [35-40].…”

The regulation effect of AMPK in immune related diseases

“…Besides, AMPK activation can suppress the production of IL-23 in human mo-DCs (Shi et al, 2016) and MEK1/2-ERK1/2-AMPK signaling axis participated in promoting survival of mDCs, suggesting that AMPK may be a potential target to modulate mDC lifespan and the immune response (López-Cotarelo et al, 2015).…”