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UFM24, a novel natural product, inhibits superoxide generation and CD11b expression in activated human neutrophils
JSID Abstracts / Journal of Dermatological Science 69 (2013) e1–e46
measurement of pruritus is difficult because pruritus is a subjective sensation. In addition, if we compare serum samples at different time points from the same patient who show dramatic improvement of skin conditions, almost all chemokine levels may well decrease. In this study, we collected serum samples twice with an eight-week interval from 10 Japanese patients with well- or moderately controlled AD. At the same time, the extent of pruritus was evaluated using visual analogue scale (VAS). We investigated serum levels of common 12 chemokines, i.e., IL-8, MCP-1, RANTES, MIP1␣, MIP-1, IP-10, I-TAC, MIG, eotaxin, TARC, MDC, and GRO␣, with enzyme-linked immunosorbent assay. There were strong correlations between serum levels of MIP-1␣ and MIP-1 and between those of MDC and TARC, consistent with previous reports. Strong correlations among serum levels of IP-10, I-TAC, and MIG were also observed. Interestingly, serum levels of eotaxin, chemoattractant for eosinophils, correlated with those of Th1 chemokines such as IP-10, I-TAC, and MIG. In addition, serum levels of GRO␣, chemoattractant for neutrophils, correlated those with MIP-1␣, IP-10, I-TAC, MIG, and eotaxin. Variations in pruritus VAS score were positively correlated with those in serum TARC levels and negatively correlated with those in serum I-TAC levels. These results suggest that TARC and I-TAC are sensitive markers reflecting minor changes in disease activity of AD and that they may contribute to pruritus in AD patients. http://dx.doi.org/10.1016/j.jdermsci.2012.11.342 P01-45 Decreased adiponectin levels in the patients with diffuse cutaneous systemic sclerosis Satoshi Fukushima ∗ , Masatoshi Jinnin, Hiroki Arakawa, Takamitsu Makino, Ikko Kajihara, Katsunari Makino, Noritoshi Honda, Yuji Inoue, Hironobu Ihn Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan Adiponectin is a protein hormone produced by visceral and subcutaneous fat cells. Adiponectin has been reported to play some roles in inflammation, angiogenesis and tissue remodeling, and induce the production of the anti-inflammatory mediators. Recently, adiponectin has also attracted attention for its involvement in the pathogenesis of collagen diseases. In this study, we determined the adiponectin expression in the sera and lesional skin of patients with systemic sclerosis (SSc). Serum adiponectin concentrations were measured in 32 patients with SSc, 10 patients with SLE, 12 patients with dermatomyositis and 13 normal subjects with specific enzyme-linked immunosorbent assays. Adiponectin mRNA was determined with real-time polymerase chain reaction in the skin tissues of 5 patients with diffuse cutaneous SSc (dcSSc), 7 patients with limited cutaneous SSc (lcSSc) and 7 normal subjects.There was significant reduction in serum adiponectin levels in patients with dcSSc compared with controls (11.2 ± 8.0 vs 139.7 ± 54.2 pg/ml, P < 0.001). SSc patients with decreased serum adiponectin levels had higher modified Rodnan total skin thickness score and higher incidence of pulmonary fibrosis, pitting scars, diffuse pigmentation or anti-topoisomerase I antibody. Adiponectin mRNA levels in skin tissues from patients with dcSSc were also reduced. In conclusions, serum adiponectin levels may be a useful biomarker for fibrotic condition in patients with SSc. Clarifying the role of adiponectin in collagen diseases may lead to further understanding of the pathogenesis and new therapeutic approach. http://dx.doi.org/10.1016/j.jdermsci.2012.11.343
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P01-46 UFM24, a novel natural product, inhibits superoxide generation and CD11b expression in activated human neutrophils Tsong-Long Hwang 1,∗ , Fang-Rong Chang 2 , Yang-Chang Wu 3 1
Graduate Institute of Natural Products, College of Medicine, Chang Gung University 2 Graduate Institute of Natural Products, Kaohsiung Medical University 3 Graduate Institute of Integrated Medicine, College of Chinese Medicine, China Medical University The oxidative stress and elastase accumulation caused by activated neutrophils play key roles in the skin aging process. The development of new agents in regulating the activations of neutrophils will be benefit for the treatment of neutrophilic skin inflammation. In this study, the anti-inflammatory effects of UFM24, a natural product, on activation of neutrophils were evaluated. Our data showed that UFM24 distinctly and concentration-dependently inhibited superoxide anion (O2 − ) generation and CD11b expression in human neutrophils induced by N-formyl-methionyl-leucyl-phenylalanine (FMLP). However, UFM24 failed to inhibit elastase release in FMLP-activated human neutrophils. Furthermore, UFM24 did not have antioxidant and superoxide-scavenging abilities. Significantly, UFM24 induced an increase in cAMP formation and protein kinase A (PKA) activity, which occurred not through an increase in adenylate cyclase (AC) but through the inhibition of cAMP-specific phosphodiesterase (PDE)4. The inhibitory effects of UFM24 on O2 − release and CD11b expression were reversed by PKA inhibitors. In addition, UFM24 reduced FMLP-induced phosphorylation of protein kinase B (AKT), but not mobilization of calcium. Significantly, the inhibitory effect of UFM24 on AKT phosphorylation was also reversed by the PKA inhibitor. In summary, these results demonstrate that the inhibition of O2 − production and CD11b expression in activated human neutrophils by UFM24 is associated with an elevation of cellular cAMP concentration and PKA activity through the inhibition of PDE4. http://dx.doi.org/10.1016/j.jdermsci.2012.11.344 P01-47 Identification of nitrotryptophan-containing proteins in the lesional skin of atopic NC/Nga mice Mitsutoshi Tominaga 1,∗ , Hiroaki Kawasaki 1 , Ayako Shigenaga 2 , Atsuko Kamo 1 , Yayoi Kamata 1 , Fumiyuki Yamakura 3 , Kenji Takamori 1,4
1
Institute for Environmental and Gender Specific Medicine, Juntendo University Graduate School of Medicine, Chiba, Japan 2 The Institute of Health and Sports Sciences, Juntendo University Graduate School of Health and Sports Science 3 School of Health Care and Nursing, Juntendo University 4 Department of Dermatology, Juntendo University Urayasu Hospital Background: Nitration of tryptophan residues is a novel posttranslational modification. This modification may be involved in many pathological processes based on regulation of protein functions and activities. Objective: This study was performed to identify the nitrotryptophan-containing proteins from the skin of NC/Nga mice, which is an animal model of atopic dermatitis (AD). Methods: Dermatitis was induced in NC/Nga mice by repeated application of Dermatophagoides farinae body (Dfb) ointment. Untreated NC/Nga and ICR mice were used as controls. Nitrotryptophan-containing proteins in the skin were detected by a combination of 2D-PAGE and Western blotting using a 6-
measurement of pruritus is difficult because pruritus is a subjective sensation. In addition, if we compare serum samples at different time points from the same patient who show dramatic improvement of skin conditions, almost all chemokine levels may well decrease. In this study, we collected serum samples twice with an eight-week interval from 10 Japanese patients with well- or moderately controlled AD. At the same time, the extent of pruritus was evaluated using visual analogue scale (VAS). We investigated serum levels of common 12 chemokines, i.e., IL-8, MCP-1, RANTES, MIP1␣, MIP-1, IP-10, I-TAC, MIG, eotaxin, TARC, MDC, and GRO␣, with enzyme-linked immunosorbent assay. There were strong correlations between serum levels of MIP-1␣ and MIP-1 and between those of MDC and TARC, consistent with previous reports. Strong correlations among serum levels of IP-10, I-TAC, and MIG were also observed. Interestingly, serum levels of eotaxin, chemoattractant for eosinophils, correlated with those of Th1 chemokines such as IP-10, I-TAC, and MIG. In addition, serum levels of GRO␣, chemoattractant for neutrophils, correlated those with MIP-1␣, IP-10, I-TAC, MIG, and eotaxin. Variations in pruritus VAS score were positively correlated with those in serum TARC levels and negatively correlated with those in serum I-TAC levels. These results suggest that TARC and I-TAC are sensitive markers reflecting minor changes in disease activity of AD and that they may contribute to pruritus in AD patients. http://dx.doi.org/10.1016/j.jdermsci.2012.11.342 P01-45 Decreased adiponectin levels in the patients with diffuse cutaneous systemic sclerosis Satoshi Fukushima ∗ , Masatoshi Jinnin, Hiroki Arakawa, Takamitsu Makino, Ikko Kajihara, Katsunari Makino, Noritoshi Honda, Yuji Inoue, Hironobu Ihn Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan Adiponectin is a protein hormone produced by visceral and subcutaneous fat cells. Adiponectin has been reported to play some roles in inflammation, angiogenesis and tissue remodeling, and induce the production of the anti-inflammatory mediators. Recently, adiponectin has also attracted attention for its involvement in the pathogenesis of collagen diseases. In this study, we determined the adiponectin expression in the sera and lesional skin of patients with systemic sclerosis (SSc). Serum adiponectin concentrations were measured in 32 patients with SSc, 10 patients with SLE, 12 patients with dermatomyositis and 13 normal subjects with specific enzyme-linked immunosorbent assays. Adiponectin mRNA was determined with real-time polymerase chain reaction in the skin tissues of 5 patients with diffuse cutaneous SSc (dcSSc), 7 patients with limited cutaneous SSc (lcSSc) and 7 normal subjects.There was significant reduction in serum adiponectin levels in patients with dcSSc compared with controls (11.2 ± 8.0 vs 139.7 ± 54.2 pg/ml, P < 0.001). SSc patients with decreased serum adiponectin levels had higher modified Rodnan total skin thickness score and higher incidence of pulmonary fibrosis, pitting scars, diffuse pigmentation or anti-topoisomerase I antibody. Adiponectin mRNA levels in skin tissues from patients with dcSSc were also reduced. In conclusions, serum adiponectin levels may be a useful biomarker for fibrotic condition in patients with SSc. Clarifying the role of adiponectin in collagen diseases may lead to further understanding of the pathogenesis and new therapeutic approach. http://dx.doi.org/10.1016/j.jdermsci.2012.11.343
e15
P01-46 UFM24, a novel natural product, inhibits superoxide generation and CD11b expression in activated human neutrophils Tsong-Long Hwang 1,∗ , Fang-Rong Chang 2 , Yang-Chang Wu 3 1
Graduate Institute of Natural Products, College of Medicine, Chang Gung University 2 Graduate Institute of Natural Products, Kaohsiung Medical University 3 Graduate Institute of Integrated Medicine, College of Chinese Medicine, China Medical University The oxidative stress and elastase accumulation caused by activated neutrophils play key roles in the skin aging process. The development of new agents in regulating the activations of neutrophils will be benefit for the treatment of neutrophilic skin inflammation. In this study, the anti-inflammatory effects of UFM24, a natural product, on activation of neutrophils were evaluated. Our data showed that UFM24 distinctly and concentration-dependently inhibited superoxide anion (O2 − ) generation and CD11b expression in human neutrophils induced by N-formyl-methionyl-leucyl-phenylalanine (FMLP). However, UFM24 failed to inhibit elastase release in FMLP-activated human neutrophils. Furthermore, UFM24 did not have antioxidant and superoxide-scavenging abilities. Significantly, UFM24 induced an increase in cAMP formation and protein kinase A (PKA) activity, which occurred not through an increase in adenylate cyclase (AC) but through the inhibition of cAMP-specific phosphodiesterase (PDE)4. The inhibitory effects of UFM24 on O2 − release and CD11b expression were reversed by PKA inhibitors. In addition, UFM24 reduced FMLP-induced phosphorylation of protein kinase B (AKT), but not mobilization of calcium. Significantly, the inhibitory effect of UFM24 on AKT phosphorylation was also reversed by the PKA inhibitor. In summary, these results demonstrate that the inhibition of O2 − production and CD11b expression in activated human neutrophils by UFM24 is associated with an elevation of cellular cAMP concentration and PKA activity through the inhibition of PDE4. http://dx.doi.org/10.1016/j.jdermsci.2012.11.344 P01-47 Identification of nitrotryptophan-containing proteins in the lesional skin of atopic NC/Nga mice Mitsutoshi Tominaga 1,∗ , Hiroaki Kawasaki 1 , Ayako Shigenaga 2 , Atsuko Kamo 1 , Yayoi Kamata 1 , Fumiyuki Yamakura 3 , Kenji Takamori 1,4
1
Institute for Environmental and Gender Specific Medicine, Juntendo University Graduate School of Medicine, Chiba, Japan 2 The Institute of Health and Sports Sciences, Juntendo University Graduate School of Health and Sports Science 3 School of Health Care and Nursing, Juntendo University 4 Department of Dermatology, Juntendo University Urayasu Hospital Background: Nitration of tryptophan residues is a novel posttranslational modification. This modification may be involved in many pathological processes based on regulation of protein functions and activities. Objective: This study was performed to identify the nitrotryptophan-containing proteins from the skin of NC/Nga mice, which is an animal model of atopic dermatitis (AD). Methods: Dermatitis was induced in NC/Nga mice by repeated application of Dermatophagoides farinae body (Dfb) ointment. Untreated NC/Nga and ICR mice were used as controls. Nitrotryptophan-containing proteins in the skin were detected by a combination of 2D-PAGE and Western blotting using a 6-