Curcuma longa enhances IFN-γ secretion by natural killer cells through cytokines secreted from macrophages.

Interferon-γ (IFN-γ) regulates the human immune system. To study the interaction between macrophages and natural killer (NK) cells, we established a THP-1 macrophage-conditioned media. Among the 58 natural plant extracts tested, Curcuma longa exerted the strongest IFN-γ-enhancing effect in NK-92 cells through THP-1 macrophages. C. longa extract (CLE) enhanced IFN-γ secretion 2.3- and 4.2-fold at 50 and 100 µg/ml, respectively. Therefore, we evaluated its IFN-γ-enhancing effect in vitro. Although NK-92 cells did not produce IFN-γ following treatment with C. longa, enhanced IFN-γ secretion was observed after treatment with THP-1 macrophage-conditioned media. We hypothesized that the cytokines secreted by the CLE-treated THP-1 macrophages are responsible for stimulating NK-92 cells. Cytokine array results show upregulation of cytokines, including MIP-1α, CXCL-1, IL-1ß, PAI-1, and TNF-α, in CLE-treated THP-1 macrophages. To determine the cytokines responsible for augmenting IFN-γ secretion, NK-92 cells were stimulated with MIP-1α, CXCL-1, IL-1ß, or PAI-1. Enzyme-linked immunosorbent assay results show that all cytokines induced IFN-γ production, although the dose response was somewhat varied. High-performance liquid chromatography analysis of CLE revealed the concentrations of three active curcuminoids, curcumin, demethoxycurcumin, and bisdemethoxycurcumin, as 6.70%, 1.00%, and 0.95%, respectively. Their mixture (with concentrations comparable to their occurrence in CLE) exerted an effect similar to that of the whole CLE. Our findings reveal that CLE indirectly stimulated NK-92 cells to secrete IFN-γ, which is mediated by cytokines produced from THP-1 macrophages. Further, we identified three curcuminoids partly responsible for this IFN-γ-enhancing effect. Therefore, C. longa can be used as a functional food ingredient owing to its immune-boosting ability. PRACTICAL APPLICATION: This study demonstrates that CLE stimulates THP-1 macrophages to secrete cytokines, which can in turn stimulate IFN-γ production by NK-92 cells. A mixture of three curcuminoids present in the extract exerted effects similar to whole CLE, demonstrating that the curcuminoids are partly responsible for the IFN-γ-enhancing effect of C. longa. Since IFN-γ is a key regulator of human immune system, these results suggest the potential use of C. longa as an immune-boosting functional food ingredient.

Adenosine and Cordycepin Accelerate Tissue Remodeling Process through Adenosine Receptor Mediated Wnt/ß-Catenin Pathway Stimulation by Regulating GSK3b Activity.

Adenosine is a cellular metabolite with diverse derivatives that possesses a wide range of physiological roles. We investigated the molecular mechanisms of adenosine and cordycepin for their promoting effects in wound-healing process. The mitochondrial energy metabolism and cell proliferation markers, cAMP responsive element binding protein 1 (CREB1) and Ki67, were enhanced by adenosine and cordycepin in cultured dermal fibroblasts. Adenosine and cordycepin stimulated adenosine receptor signaling via elevated cAMP. The phosphorylation of mitogen-activated protein kinase kinase (MEK) 1/2, mammalian target of rapamycin (mTOR) and glycogen synthase kinase 3 beta (Gsk3b) and Wnt target genes such as bone morphogenetic protein (BMP) 2/4 and lymphoid enhancer binding factor (Lef) 1 were activated. The enhanced gene expression by adenosine and cordycepin was abrogated by adenosine A2A and A2B receptor inhibitors, ZM241385 and PSH603, and protein kinase A (PKA) inhibitor H89, indicating the involvement of adenosine receptor A2A, A2B and PKA. As a result of Wnt/ß-catenin pathway activation, the secretion of growth factors such as insulin-like growth factor (IGF)-1 and transforming growth factor beta (TGFß) 3 was increased, previously reported to facilitate the wound healing process. In addition, in vitro fibroblast migration was also increased, demonstrating their possible roles in facilitating the wound healing process. In conclusion, our data strongly demonstrate that adenosine and cordycepin stimulate the Wnt/ß-catenin signaling through the activation of adenosine receptor, possibly promoting the tissue remodeling process and suggest their therapeutic potential for treating skin wounds.

Anti-inflammatory effect of Salsola komarovii extract with dissociated glucocorticoid activity.

BACKGROUND: Glucocorticoids (GCs) are anti-inflammatory drugs widely used to treat acute and chronic inflammatory diseases. However, despite their excellent efficacy, the long-term use of GCs is relatively limited owing to their adverse effects. Recent studies have sought to reduce these adverse effects by developing dissociated GCs that bind to GC receptors (GRs) to induce potent anti-inflammatory effects without the transcription of GC response element (GRE)-promoted genes. Some species of the genus Salsola are used in traditional Chinese medicine to treat cancer, hypertension, and inflammation. In this study, we investigated the potential dissociated GC activities and underlying mechanisms of Salsola komarovii (SK), which is native to Korea. METHODS: To determine whether SK ethanol extract (SEE) directly interacts with the GR, an in vitro fluorescence polarization based-GR competitor assay was performed. The effect of SEE on the transcriptional activity of nuclear factor (NF)-κB and GRE was confirmed in HepG2 cells using the Cignal reporter assay. The anti-inflammatory effect of SK was determined by assessing lipopolysaccharide (LPS)-induced interleukin (IL)-6 production. To confirm whether SEE induces GRE-driven gene expression, preadipocyte differentiation followed by lipid deposition was performed in the presence of SEE. RESULTS: SEE exhibited GR binding activity in the fluorescence polarization competitive binding assay and induced GR nuclear translocation. It also interfered with the nuclear translocation of NF-κB and the NF-κB-dependent transcriptional activity based on the immunofluorescence analysis and reporter assay, respectively. SEE exerted anti-inflammatory effects by reducing LPS-induced IL-6 production as effectively as hydrocortisone (positive control). SK did not induce GRE-driven gene expression and preadipocyte differentiation, which is one of the major adverse effects of GCs. CONCLUSIONS: Collectively, these results suggest that SK could be a novel and safe anti-inflammatory agent with dissociated GC properties and, therefore, it has great potential for use in treating inflammatory disorders.

Polygonum multiflorum extract support hair growth by elongating anagen phase and abrogating the effect of androgen in cultured human dermal papilla cells.

BACKGROUND: Dermal papilla cells (DPCs) play a key role in hair growth among the various cell types in hair follicles. Especially, DPCs determine the fate of hair follicle such as anagen to telogen transition and play a pivotal role in androgenic alopecia (AGA). This study was performed to elucidate the hair growth promoting effects of Polygonum multiflorum extract (PM extract) in cultured human DPCs and its underlying mechanisms. METHODS: The effects of PM extract on cultured DPCs were investigated. Cell viability and mitochondrial activity were measured by CCK-8 and JC-1 analysis, respectively. Western blotting, dot blotting, ELISA analysis, immunocytochemistry and real-time PCR analysis were also performed to elucidate the changes in protein and mRNA levels induced by PM extract. 3D cultured DPC spheroids were constructed for mimicking the in vivo DPs. The hair growth stimulatory effect of PM extract was evaluated using human hair follicle organ culture model. RESULTS: PM extract increased the viability and mitochondrial activity in cultured human DPCs in a dose dependent manner. The expression of Bcl2, an anti-apoptotic protein expressed dominantly in anagen was significantly increased and that of BAD, a pro-apoptotic protein expressed in early catagen was decreased by PM extract in cultured DPCs and/or 3D DPC spheroid culture. PM extract also decreased the expression of catagen inducing protein, Dkk-1. Growth factors including IGFBP2, PDGF and VEGF were increased by PM extract, revealed by dot blot protein analysis. We also have found that PM extract could reverse the androgenic effects of dihydrotestosterone (DHT), the most potent androgen. Finally, PM extract prolonged the anagen of human hair follicles by inhibiting catagen entry in human hair follicle organ culture model. CONCLUSION: Our data strongly suggest that PM extract could promote hair growth by elongating the anagen and/or delaying the catagen induction of hair follicles through activation of DPCs.

Hair Growth Promoting Effect of Hottuynia cordata Extract in Cultured Human Hair Follicle Dermal Papilla Cells.

Houttuynia cordata (HC) is a traditional oriental herbal medicinal plant widely used as a component of complex prescriptions in Asia for alopecia treatment. The effect of HC on hair growth and its underlying mechanism, however, have not been demonstrated or clarified. In this study, we investigated the hair growth promoting effect of HC in cultured human dermal papilla cells (hDPCs). HC extract was found to stimulate the proliferation of hDPCs and this stimulation might be in part a consequence of activated cellular energy metabolism, because treatment of HC extract increased the generation of nicotinamide adenine dinucleotide (NADH) and ATP through increasing the mitochondrial membrane potential (ΔΨ). In the context of cell cycle, HC extract increased the expression of CDK4 and decreased the expression of CCNA2 and CCNB1, implying that HC extract might induce G1 phase progression of DPCs which resulted in enhanced proliferation. HC extract increased the expression of Bcl2 essential for maintaining hair follicle anagen stage and cell survival. On the contrary, the expression of p16 and p21 was down-regulated by HC extract. In addition, HC extract enhanced the secretion of platelet-derived growth factor (PDGF)-aa and vascular endothelial growth factor (VEGF) and induced phosphorylation of extracellular signal-regulated kinase (ERK) and AKT. Furthermore, HC extract prolonged anagen stage in organ cultured human hair follicles. Our data strongly suggest that HC extract could support hair growth by stimulating proliferation of DPCs and elongating anagen stage, resulted from enhanced cellular energy metabolism and modulation of gene expression related to cell cycle, apoptosis, and growth factors.

Erjingwan Extracts Exert Antiaging Effects of Skin through Activating Nrf2 and Inhibiting NF-κB.

In oriental medicine, mixtures of medical plants are always used as prescriptions for diseases. Natural products extracted from herbs have great potential antiaging effects. Previous studies and clinical trials have shown several critical functions of Erjingwan (EJW), such as nourishing Yin, kidney tonifying and aging-resistance. We assumed that EJW extracts exerted the antiaging effects through nourishing Yin. We examined the antiaging effects of EJW extracts on healthy human skin by noninvasive measurements. Then we estimated the cell proliferation and DPPH radical scavenging rate. Western blotting analysis was used to determine the expressions of matrix metalloproteinase-1 (MMP-1), type I collagen (COL1A2), p-NF-κB, NF-κB, p-IκBα, IκBα, p-Nrf2, and HO-1. EJW extracts did not affect moisture content, TEWL and skin chroma, while it significantly improved skin glossiness and skin elasticity. Moreover, EJW extracts could downregulate the MMP1 expression and upregulate the COL1A2 expression. In addition, it promoted the Nrf2 pathway while it inhibited the NF-κB pathway. With the application of cream containing EJW extracts, the skin aging state was significantly improved. Furthermore, in vitro studies showed that EJW extracts contributed to the repair of skin after injury. Taken together, the antiaging effects of EJW extracts were related to its antioxidant and anti-inflammatory abilities.

Clinical efficacy of herbal extract cream on the skin hydration, elasticity, thickness, and dermis density for aged skin: A randomized controlled double-blind study.

BACKGROUND: Traditional medicine herbal prescriptions used for the treatment of skin disease have been developed into cosmetics. Sang-Hyul-Yun-Boo-Em (SHYBE) is a mixed herbal formula prescribed for patients with yin or blood deficiency patterns of skin disease. A previous study reported that SHYBE exercises anti-allergic and anti-inflammatory effects. To date, no study has reported the efficacy of cosmetics containing the SHYBE extract. AIMS: To observe the efficacy of SHYBE extract cream on hydration, elasticity, thickness, and dermis density in aged skin. METHODS: This was a double-blind randomized placebo-controlled parallel-group trial. The trial consisted of an 8-week topical application of the test or placebo products with two visits at 4-week intervals. A total of 46 healthy Korean females, aged 40-59, were enrolled in this study. Objective skin assessments for hydration, elasticity, thickness and dermis density, self-assessment, and safety assessment were conducted. RESULTS: Sang-Hyul-Yun-Boo-Em extract cream improved skin hydration, elasticity, and dermal density in Asian middle-aged females compared with placebo cream, which excluded SHYBE extract and contained other cosmetic materials. CONCLUSIONS: Sang-Hyul-Yun-Boo-Em extract cream showed anti-aging properties in middle-aged women. It could be recommended for aging skin with dryness, and loss of elasticity and density.

Anti-allergic inflammatory effect of vanillic acid through regulating thymic stromal lymphopoietin secretion from activated mast cells.

Vanillic acid, which is well known as a benzoic acid derivative, has been used as a flavouring agent. Currently, we ascertained the therapeutic potential action of vanillic acid on allergic inflammatory reaction in human mast cell line, HMC-1. Treatment with vanillic acid resulted in a significant decrease in levels of thymic stromal lymphopoietin and pro-inflammatory cytokines compared to phorbol 12-myristate 13-acetate and calcium ionophore A23187 (PMACI)-treated HMC-1 cells. In PMACI-stimulated cells, treatment with vanillic acid also dramatically inhibited activities of caspase-1 and nuclear factor-kB (p65). Furthermore, treatment with vanillic acid suppressed phosphorylation of mitogen-activated protein kinases in PMACI-treated HMC-1 cells. Taken together, these findings suggest that vanillic acid has a beneficial effect on allergic inflammatory disorders.

Ameliorative effect of atractylenolide III in the mast cell proliferation induced by TSLP.

Atractylenolide III (ATL-III) is an active compound of Atractylodes lancea, which has been widely used for the treatment of cancer. Cancer is closely connected with inflammation, and many anti-inflammatory agents are also used to treat cancer. We investigated the influence of ATL-III on thymic stromal lymphopoietin (TSLP)-induced inflammatory reactions. Pretreatment with ATL-III suppressed murine double minute 2 levels and promoted p53 levels in TSLP-treated human mast cell, HMC-1 cells. Mast cell proliferation increased by TSLP or IL-3 stimulation was significantly decreased by ATL-III pretreatment. Interleukin (IL)-13 and phosphorylated signal transducer and activator of transcription 3, 5, and 6 levels in TSLP-treated HMC-1 cells were also decreased by ATL-III pretreatment. In addition, ATL-III decreased the TSLP-induced production of proinflammatory cytokines (IL-6, IL-1ß, tumor necrosis factor-α, and IL-8). ATL-III decreased the levels of Bcl2 and procaspase-3 and increased caspase-3 activation and cleaved PARP levels. Furthermore, ATL-III decreased TSLP-induced mast cell proliferation and the production of inflammatory cytokine by LAD2 cells. Taken together, these findings suggest that ATL-III plays a useful role as an anti-inflammatory agent and should be viewed as a potential anti-cancer agent.

Cordycepin Suppresses Thymic Stromal Lymphopoietin Expression via Blocking Caspase-1 and Receptor-Interacting Protein 2 Signaling Pathways in Mast Cells.

Cordycepin (3'-deoxyadenosine) is one of the active components isolated from Cordyceps militaris, and has been shown to have anti-inflammatory, anti-oxidant, anti-aging, and anti-cancer effects. Mast cell-derived thymic stromal lymphopoietin (TSLP) plays an important role in the pathogenesis of allergic inflammatory reactions. Here, we investigated the regulatory effect and mechanisms of cordycepin on the expression of TSLP in the human mast cell line, HMC-1 cells, and in the human keratinocyte cell line, HaCaT cells. Cordycepin significantly decreased the production and mRNA expression of TSLP through the inhibition of caspase-1 and nuclear factor-κB activation. Cordycepin also significantly reduced the phosphorylation of receptor-interacting protein 2 and inhibitory kappa B (IκB) kinase ß. Cordycepin significantly decreased the production and mRNA expression of interleukin (IL)-8, IL-1ß, IL-6, and tumor necrosis factor-α in activated HMC-1 cells. Moreover, cordycepin significantly decreased the levels of TSLP in activated HaCaT cells. Our studies suggest that cordycepin can be applied to the treatment of allergic inflammatory diseases exacerbated by TSLP.

Inhibition of c-Kit signaling by diosmetin isolated from Chrysanthemum morifolium.

The interaction of stem cell factor (SCF) with its cognate receptor c-Kit is closely associated with the survival and maturation of melanocytes. To investigate novel depigmentation agents, we screened 2,000 plant extracts for c-Kit inhibitors to identify active small molecules by using time-resolved fluorescence enzyme assays. For the active extracts identified as inhibitors of c-Kit enzyme, we evaluated the effects of the active extracts and isolated flavonoids on c-Kit phosphorylation in MO7e/melanocytes. Anti-melanogenic activity was also examined in melanocytes and melanoderm model. The flavonoids such as diosmetin, apigenin, acacetin and luteolin isolated from Chrysanthemum morifolium were found to be active in inhibiting c-Kit both at enzyme and cellular levels. In addition, these flavonoids attenuated SCF-induced proliferation of human primary melanocytes without toxicity and suppressed ultraviolet (UV) B irradiation-mediated melanin synthesis significantly. Among the active flavonoids, diosmetin was found to inhibit SCF-induced melanogenesis in a human melanoderm model. These results strongly suggest that C. morifolium extract and diosmetin have potential to suppress SCF-/UVB-induced melanogenesis, and could be developed as anti-pigmentation agents.

Cedrol Enhances Extracellular Matrix Production in Dermal Fibroblasts in a MAPK-Dependent Manner.

BACKGROUND: The extracellular matrix (ECM) produced by dermal fibroblasts supports skin structure, and degradation and/or reduced production of ECM are the main causes of wrinkle formation. OBJECTIVE: The aim of this study was to identify the active ingredient that enhances ECM production in dermal fibroblasts. METHODS: Polarity-based fractionation was used to isolate the active ingredient from natural extracts, and the effects of cedrol (isolated from Pterocarpus indicusirginia) on ECM production in cultured human dermal fibroblasts was investigated by reverse transcription-polymerase chain reaction, enzyme linked immunosorbent assay, and Western blot analysis. RESULTS: Cedrol accelerated fibroblast growth in a dose-dependent manner and increased the production of type 1 collagen and elastin. Phosphorylation of p42/44 extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, and Akt was markedly increased by cedrol, indicating that enhanced ECM production is linked to activation of intracellular signaling cascades. CONCLUSION: These results indicate that cedrol stimulates ECM production, with possible applications to the maintenance of skin texture.

Inhibition of melanogenesis by piceid isolated from Polygonum cuspidatum.

Piceid (5,4'-dihydroxystilbene-3-O-ß-D-glucopyranoside) is one of the stilbenes found in Polygonum cuspidatum. Previous studies have shown that this compound has little effect on tyrosinase inhibition when compared with other stilbenes in a cell-free tyrosinase assay. Furthermore, its role for melanogenesis in melanocytes is relatively unknown. In melanocytes, piceid inhibits tyrosinase activity and melanin production in a concentration-dependent manner. To explore the action of piceid on melanogenesis, we studied its effect on several key cellular enzymes and a transcriptional factor known to be involved in melanogenesis, including: tyrosinase, tyrosinase-related protein 1, tyrosinase-related protein 2, and microphthalmia-associated transcription factor. Interestingly, the effects of piceid on hypopigmentation and inhibition of tyrosinase activity were better than those of arbutin, which is well known to inhibit melanin formation in melanocytes. In addition, piceid suppressed the mRNA and protein expression of the aforementioned enzymes and transcriptional factor in a concentration-dependent manner. In this regards, our results showed that piceid represents a safe and new candidate for a skin-lightening agent.

Stimulation of the extracellular matrix production in dermal fibroblasts by velvet antler extract.

BACKGROUND: Fibroblasts produce many components of the extracellular matrix (ECM) and so they contribute to the maintenance of connective tissue integrity. OBJECTIVE: The aim of this study is to evaluate the effect of velvet antler extract (VAE) on the ECM production of dermal fibroblasts cultured in vitro. METHODS: Primary cultured human dermal fibroblasts were treated with VAE, and then the ECM production was determined by RT-PCR, ELISA and Western blot analysis. Furthermore, the change of gene expression according to VAE treatment was evaluated by cDNA microarray. RESULTS: VAE accelerated the growth of fibroblasts in a dose-dependent manner. VAE increased the production of several ECM components, including type 1 collagen, fibronectin and elastin. In line with these results, the phosphorylations of p42/44 ERK and p38 mitogen-activated protein kinase were markedly increased by VAE, suggesting that the enhancement of ECM production may be linked to the activation of intracellular signaling cascades. VAE also significantly increased cell migration on an in vitro scratch wound test. In cDNA microarray, many genes related with connective tissue integrity were identified to be up-regulated by VAE. CONCLUSION: These results suggest that VAE has a potential to stimulate ECM production, and VAE may be applicable for maintaining the skin's texture.

Partially purified paeoniflorin exerts protective effects on UV-induced DNA damage and reduces facial wrinkles in human skin.

Partially purified paeoniflorin (PF), a new cosmetic ingredient from roots of Paeoniae lactiflora, has been developed. Its paeoniflorin content is about 64%, far higher than that of conventional, cosmetic-grade peony root extracts (approximately 10%). In this report, we studied the effects of PF on UV-induced DNA damage in both cultured human keratinocytes and hairless mouse skin. We also investigated the anti-wrinkle effects of PF-containing cosmetic preparations on human skin. From the in vitro and in vivo comet assay, it was revealed that PF protected cells from DNA damage induced by ultraviolet B (UVB) irradiation in both cultured normal human keratinocytes (19.4% decrease at 0.001%) and hairless mouse skin keratinocytes (41% decrease at 0.01%). An eight-week clinical trial using 0.5% PF-containing formulation with 20 volunteers resulted in a statistically significant reduction in facial wrinkles (p < 0.05). These results suggest that the partially purified paeoniflorin has potent anti-aging and anti-wrinkle activities and should be a useful ingredient for these purposes.