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Fig 1.

Effect of hemp seed hexane extracts against the growth of P. acnes.

All data are presented as mean±SD of three independent experiments. *p<0.05 indicates significant differences compared to control group.

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Fig 2.

Inhibitory effects of hemp seed hexane extracts on P. acnes-induced inflammation in HaCaT cells.

(A) Effects of hemp seed hexane extracts (HSHE) on iNOS, COX-2 and IL-1β expression in P. acnes-stimulated HaCaT cells. The expressions of iNOS, COX-2, and IL-1β were analyzed with ImageJ and normalized against β-actin. (B) Effects of HSHE on NO production in P. acnes-stimulated HaCaT cells. ELISA results demonstrate that HSHE reduced PGE2 (C) and IL-8 (D) in P. acnes-infected HaCaT cells. ELISA results demonstrate that HSHE reduced in P. acnes-infected HaCaT cells. All data are expressed as mean ± SD. *P<0.05 compared with P. acnes treated cells only.

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Fig 3.

Hemp seed hexane extracts effectively inhibit the NF-κB signaling pathway in P. acnes-treated HaCaT cells.

The expression of p-IKKα/β, p-IκB-α and p-NF-κB were analyzed with ImageJ and normalized against β-actin. Results are expressed as mean ± SD. *P<0.05 compared with P. acnes treated cells only.

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Fig 4.

Effect of hemp seed hexane extracts on p-NFκB p65 nuclear translocation in P. acnes-induced HaCaT cells.

Immunofluorescence staining for p-NF-κB (red) in P. acnes-exposed HaCaT cells without and with 0.6% HSHE. Nuclei are stained with DAPI (blue). HSHE reduced the nuclear translocation and accumulation of p-NF-κB, which was induced by P. acnes. Scale bar = 20 μm.

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Fig 5.

Hemp seed hexane extracts effectively inhibit the MAPK signaling pathway in P. acnes-treated HaCaT cells.

(A) The expression of p-p38, p-ERK, and p-JNK were analyzed with ImageJ and normalized against β-actin. (B) Transcriptional activation of AP-1 was analyzed by luciferase reporter assay. AP-1 signaling was down-regulated by HSHE in HaCaT cells. Results are expressed as mean ± SD. *P<0.05 compared with P. acnes treated cells only.

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Fig 6.

Stimulatory effect of hemp seed hexane extracts on in vitro collagen synthesis.

(A) Effect of HSHE on collagen production in P. acnes-induced Hs68 cells. (B) Effect of HSHE on P. acnes-induced MMP-9 activity (gelatinase) on zymographic gel. The expression of MMP-9 was analyzed with ImageJ and normalized against P. acnes treated cells only. All data are expressed as mean ± SD. *P<0.05 compared with P. acnes treated cells only.

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Fig 7.

Regulation of intracellular lipogenesis through AMPK signals in IGF-1-induced sebocytes.

The expression of p-AMPKα, p-mTOR, SREBP1, and FAS was analyzed with ImageJ and normalized against β-actin. Results are expressed as mean ± SD. *P<0.05 compared with IGF-1 treated cells only.

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Fig 8.

Regulation of intracellular lipogenesis through the AKT/FoxO1 signals in IGF-1-induced sebocytes.

The expression levels of p-AKT and p-FoxO1 were analyzed with ImageJ and normalized against β-actin. Results are expressed as mean ± SD. *P<0.05 compared with IGF-1 treated cells only.

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Fig 9.

Inhibitory effect of hemp seed hexane extracts on the IGF-1-stimulated 5-lipoxygenase level in sebocytes.

All data are expressed as mean ± SD. *P<0.05 compared with IGF-1 treated cells only.

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Fig 10.

The underlying mechanism for anti-acne activity of hemp seed hexane extracts via anti-inflammation in P.acnes-stimulated HaCaT cells and anti-lipogenesis in sebocytes.

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