Curcumin Improves Keratinocyte Proliferation, Inflammation, and Oxidative Stress through Mediating the SPAG5/FOXM1 Axis in an In Vitro Model of Actinic Dermatitis by Ultraviolet

Figure 3

Overexpression of SPAG5 potentiated UV irradiation-mediated dysfunction of keratinocytes. (a) Vectors and SPAG5 overexpression plasmids were transfected into HaCaT cells, and the SPAG5 expression in HaCaT cells was assayed by RT-PCR 24 hours later. The transfected HaCaT cells were irradiated with UV and then treated with curcumin (5 μM) for 24 hours. (b) CCK-8 was applied to check the proliferation of UV-irradiated HaCaT cells. (c)–(f) Levels of SOD, GSH-PX, MDA, and ROS in UV-irradiated HaCaT cells were assessed using an oxidative stress assay kit and cytofluorimetry. (g) ELISA tested the expression of inflammatory factors IL-1β, IL-6, IL-18, and TNFα in UV-irradiated HaCaT cells. (h)–(j) WB was conducted to investigate the profiles of apoptosis-associated proteins (Bax, Bcl-xL, Caspase3, Caspase8, Caspase9), oxidative stress-associated proteins (Keap1, Nrf2, HO-1, COX2, iNOS), and inflammatory response proteins (NF-κB, MMP1, MMP). (k) The expression of the SPAG5/FOXM1 pathway in UV-irradiated HaCaT cells was estimated by WB. . (vs. vector or UV group), , ; ^# (vs. SPAG5+UV group), ^##, ^###.

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