A Water-Soluble Extract from Actinidia arguta Ameliorates Psoriasis-Like Skin Inflammation in Mice by Inhibition of Neutrophil Infiltration

Hyun-Keun Kim, Min Jung Bae, Seonung Lim, Wonwoo Lee, Sunyoung Kim, Hyun-Keun Kim, Min Jung Bae, Seonung Lim, Wonwoo Lee, Sunyoung Kim

Abstract

Psoriasis is a chronic inflammatory disease with complex etiology involving multiple factors. Current treatment methods are highly limited and there is a strong need for the development of safer and efficacious agents. We have previously shown that a water-soluble extract derived from hardy kiwifruit Actinidia arguta, called PG102, shows potent anti-inflammatory effects. Based on its reported biological activities, the effects of PG102 were examined on imiquimod-induced psoriasis-like skin inflammation. Our results showed that topical application of PG102 ameliorates clinical symptoms of psoriasis, reducing skin thickness and Interleukin (IL)-17A level in draining lymph nodes without causing any adverse effects. Treatment with PG102 on cytokine-stimulated HaCaT cells suppressed hyperproliferation and downregulated the expression of various chemokines and antimicrobial peptides known to induce neutrophil infiltration. These anti-inflammatory activities of PG102 were mediated via inhibition of NF-κB and signal transducer of activation (STAT) signaling. We also found decreased neutrophil chemotaxis both in vitro and in vivo. Taken together, PG102 has potential as a safe and effective reagent for the treatment of psoriasis.

Keywords: Actinidia arguta; PG102; keratinocyte; neutrophil; psoriasis.

Conflict of interest statement

M.J.B., W.L. and S.K. are the employees of ViroMed Co.Ltd. S.K. owns stocks of this company. Other authors have no conflict of interest.

Figures

Figure 1
Figure 1
PG102 is a standardized extract from Actinidia arguta. (A) Quantification of marker compounds citric acid and quinic acid; (B) Interleukin (IL)-8 bioassay and half maximal inhibitory concentration (IC50) of PG102 in HaCaT cells. Ref: reference. RT: retention time.
Figure 2
Figure 2
Topical treatment of PG102 alleviates imiquimod (IMQ)-induced psoriasis-like symptoms. (A) Experimental scheme (B) Photo of dorsal skin of Control (DMSO only), Vehicle (IMQ + DMSO), PG102 (IMQ + PG102 100 mg/kg) and Dexamethasone (IMQ + Dex 1 mg/kg) treated mice. (C) Psoriasis Area and Severity Index (PASI) scoring. (D) Hematoxylin-eosin (H&E) staining of dorsal skin (200×). (E) Interleukin (IL)-17A protein level from phorbol 12-myristate 13-acetate (PMA)/ionomycin-restimulated cells isolated from draining lymph nodes. (F) Change in body weight. Representative results from at least three independent experiments are shown (n = 3–4). The data are shown as the mean ± standard error of mean (SEM). ##p < 0.01, ####p < 0.0001 versus Control group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus vehicle group.
Figure 2
Figure 2
Topical treatment of PG102 alleviates imiquimod (IMQ)-induced psoriasis-like symptoms. (A) Experimental scheme (B) Photo of dorsal skin of Control (DMSO only), Vehicle (IMQ + DMSO), PG102 (IMQ + PG102 100 mg/kg) and Dexamethasone (IMQ + Dex 1 mg/kg) treated mice. (C) Psoriasis Area and Severity Index (PASI) scoring. (D) Hematoxylin-eosin (H&E) staining of dorsal skin (200×). (E) Interleukin (IL)-17A protein level from phorbol 12-myristate 13-acetate (PMA)/ionomycin-restimulated cells isolated from draining lymph nodes. (F) Change in body weight. Representative results from at least three independent experiments are shown (n = 3–4). The data are shown as the mean ± standard error of mean (SEM). ##p < 0.01, ####p < 0.0001 versus Control group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus vehicle group.
Figure 3
Figure 3
PG102 suppresses hyperproliferation of keratinocytes through STAT3 signaling. (A) WST-1 cell proliferation assay in HaCaT cells treated with M5 and PG102 for 24 h and 48 h (n = 3). (B) PG102 was treated to M5-stimulated HaCaT cells for 30 min and Western blot analysis was performed. (C) Densitometry results of Western blot. The means of three independent experiments are shown. p-STAT3 (Y705), phospho-signal transducer and activator of transcription 3 (Tyr705). p-STAT3 (S727), phospho-signal transducer and activator of transcription 3 (phospho S727). ##p < 0.01, ####p < 0.0001 versus Control group; * p < 0.05, ** p < 0.01 versus M5-only treated group. The data are shown as the mean ± standard error (SD).
Figure 3
Figure 3
PG102 suppresses hyperproliferation of keratinocytes through STAT3 signaling. (A) WST-1 cell proliferation assay in HaCaT cells treated with M5 and PG102 for 24 h and 48 h (n = 3). (B) PG102 was treated to M5-stimulated HaCaT cells for 30 min and Western blot analysis was performed. (C) Densitometry results of Western blot. The means of three independent experiments are shown. p-STAT3 (Y705), phospho-signal transducer and activator of transcription 3 (Tyr705). p-STAT3 (S727), phospho-signal transducer and activator of transcription 3 (phospho S727). ##p < 0.01, ####p < 0.0001 versus Control group; * p < 0.05, ** p < 0.01 versus M5-only treated group. The data are shown as the mean ± standard error (SD).
Figure 4
Figure 4
PG102 downregulates expression of chemokines on M5-stimulated HaCaT cells in a dose-dependent manner. (A) Real-time polymerase chain reaction (PCR) (qPCR) analysis of mRNA expression levels of chemokines after 6 h. (B) Protein levels of chemokines measured in the cultured supernatant by enzyme-linked immunosorbent assay (ELISA) after 24 h. CXCL, chemokine (C-X-C motif) ligand. ####p < 0.001 versus negative control group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus M5-only treated group.
Figure 5
Figure 5
PG102 downregulates expression of antimicrobial peptides on M5-stimulated HaCaT cells in a dose-dependent manner. (A) Real-time PCR (qPCR) analysis of mRNA expression levels of antimicrobial peptides after 24 h. (B) Protein levels of antimicrobial peptides measured in the cultured supernatant by ELISA after 48 h. BD-2, beta-defensin 2; S100, S100 calcium binding protein. ####p < 0.0001 versus negative control group; * p < 0.05, **** p < 0.0001 versus M5-only treated group.
Figure 6
Figure 6
PG102 inhibits NF-κB and STAT1 signaling in HaCaT cells. (A) Western blot results of corresponding proteins after treatment with M5 and PG102 for 30 min. (B) Densitometry of Western blot. The means of three experiments are shown. ###p < 0.001, ####p < 0.0001 versus Control group; * p < 0.05 versus M5-only treated group.
Figure 7
Figure 7
PG102 suppresses neutrophil chemotaxis in vitro and in vivo. (A) Chemotaxis assay of differentiated HL-60 (dHL-60) cells. (B) IHC staining for Ly6G in dorsal skin of control, vehicle, PG102 (100 mg/kg), Dex (1 mg/kg) treated mice. (C) Quantification of Ly6G+ cells per five random high power fields (400X). (D) Real-time qPCR analysis of mRNA expression levels of antimicrobial peptides and chemokines in dorsal skin of mice (n = 3–4). FBS, fetal bovine serum; Dex, dexamethasone; M5, mixture of 5 cytokines. The data are shown as the mean ± SEM. ####p < 0.0001 versus Control group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus vehicle group.

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