ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Psoralea corylifolia L. (PC) is widely used in traditional medicines to treat inflammatory and infectious diseases. Isobavachin (IBC) is a bioavailable prenylated flavonoid derived from PC that has various biological properties. However, little information is available on its anti-inflammatory effects and mechanisms of action. AIM OF THE STUDY: In this study, we aimed to determine the anti-inflammatory effects of IBC in vitro and in vivo by conducting a mechanistic study using murine macrophages. MATERIALS AND METHODS: We evaluated the modulatory effects of IBC on the production of pro-inflammatory cytokines and mediators in murine macrophages. In addition, we examined whether IBC inhibits lipopolysaccharide (LPS)-induced inflammatory responses in a zebrafish model. Alterations in inflammatory response-associated genes and proteins were determined using quantitative reverse transcriptional polymerase chain reaction (RT-qPCR) and Western blotting analysis. RESULTS: IBC markedly reduced the overproduction of inflammatory mediators, pro-inflammatory cytokines, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), phosphorylation of mitogen-activated protein kinase (MAPK) and nuclear translocation of nuclear factor-kappa B (NF-κB) in macrophages induced by lipopolysaccharides (LPS). In addition, excessive NO, ROS, and neutrophil level induced by LPS, were suppressed by IBC treatment in a zebrafish inflammation model. CONCLUSIONS: Collectively, bioavailable IBC inhibited on the inflammatory responses by LPS via MAPK and NF-κB signaling pathways in vitro and in vivo, suggesting that it may be a potential modulatory agent against inflammatory disorders.
Subject(s)
Mitogen-Activated Protein Kinases , Psoralea , Animals , Mice , Mitogen-Activated Protein Kinases/metabolism , NF-kappa B/metabolism , Lipopolysaccharides/pharmacology , Zebrafish , Psoralea/metabolism , Signal Transduction , Flavonoids/pharmacology , Cytokines/metabolism , Macrophages , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Anti-Inflammatory Agents/metabolism , Nitric Oxide Synthase Type II/metabolism , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolismABSTRACT
The incidence of ulcerative colitis (UC), an inflammatory disorder of the gastrointestinal tract, has rapidly increased in Asian countries over several decades. To overcome the limitations of conventional drug therapies, including biologics for UC management, the development of herbal medicine-derived products has received continuous attention. In this study, we evaluated the beneficial effects of a hydroethanolic extract of Fritillariae thunbergii Bulbus (FTB) in a mouse model of DSS-induced UC. The DSS treatment successfully induced severe colonic inflammation and ulceration. However, the severity of colitis was reduced by the oral administration of FTB. Histopathological examination showed that FTB alleviated the infiltration of inflammatory cells (e.g., neutrophils and macrophages), damage to epithelial and goblet cells in the colonic mucosal layer, and fibrotic lesions. Additionally, FTB markedly reduced the gene expression of proinflammatory cytokines and extracellular matrix remodeling. Immunohistochemical analysis showed that FTB alleviated the decrease in occludin and zonula occludens-1 expression induced by DSS. In a Caco-2 monolayer system, FTB treatment improved intestinal barrier permeability in a dose-dependent manner and increased tight junction expression. Overall, FTB has potential as a therapeutic agent through the improvement of tissue damage and inflammation severity through the modulation of intestinal barrier integrity.
Subject(s)
Colitis, Ulcerative , Colitis , Humans , Animals , Mice , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/metabolism , Dextran Sulfate/toxicity , Caco-2 Cells , Intestines/pathology , Colitis/chemically induced , Inflammation/metabolism , Colon/metabolism , Intestinal Mucosa/metabolism , Disease Models, Animal , Mice, Inbred C57BLABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: In traditional oriental medicine, the dried seeds of Psoralea corylifolia L. (PC) have been used to treat various diseases, including gastrointestinal, urinary, orthopedic, diarrheal, ulcer, and inflammatory disorders. AIM OF THE STUDY: Although its various biological properties are well-known, there is no information on the therapeutic effects and bioavailable components of PC against inflammatory bowel disease. Therefore, we focused on the relationship between hydroethanolic extract of PC (EPC) that ameliorates colitis in mice and bioactive constituents of EPC that suppress pro-inflammatory cytokines in macrophages. MATERIALS AND METHODS: We investigated the therapeutic effects of EPC in a dextran sulfate sodium-induced colitis mouse model and identified the orally absorbed components of EPC using UPLC-MS/MS analysis. In addition, we evaluated and validated the mechanism of action of the bioavailable constituents of EPC using network pharmacology analysis. The effects on nitric oxide (NO) and inflammatory cytokines were measured by Griess reagent and enzyme linked immunosorbent assay in lipopolysaccharide (LPS)-induced macrophages. RESULTS: In experimental colitis, EPC improved body weight loss, colon length shortening, and disease activity index. Moreover, EPC reduced the serum levels of pro-inflammatory cytokines and histopathological damage to the colon. Network pharmacological analysis identified 13 phytochemicals that were bioavailable following oral administration of EPC, as well as their potential anti-inflammatory effects. 11 identified EPC constituents markedly reduced the overproduction of NO, tumor necrosis factor-α, and/or interleukin-6 in macrophages induced by LPS. The LPS-induced expression of the nuclear factor kappa-light-chain-enhancer of activated B cells reporter gene was reduced by the 4 EPC constituents. CONCLUSIONS: The results indicate that the protective activity of EPC against colitis is a result of the additive effects of each constituent on the expression of inflammatory cytokines. Therefore, it suggests that 11 bioavailable phytochemicals of EPC could aid in the management of intestinal inflammation, and also provides useful insights into the clinical application of PC for the treatment of inflammatory bowel diseases.
Subject(s)
Colitis, Ulcerative , Colitis , Fabaceae , Psoralea , Mice , Animals , Psoralea/metabolism , Lipopolysaccharides/pharmacology , Chromatography, Liquid , Network Pharmacology , Tandem Mass Spectrometry , Colitis/chemically induced , Colitis/drug therapy , Colitis/pathology , Anti-Inflammatory Agents/adverse effects , Colon , Cytokines/metabolism , Dextran Sulfate , Colitis, Ulcerative/drug therapy , NF-kappa B/metabolismABSTRACT
Natural and synthetic chalcones exhibit anti-inflammatory, antitumoral, antibacterial, antifungal, antimalarial, and antitubercular activities. Isodorsmanin A (IDA), a chalcone, is a well-known constituent of the dried seeds of Psoralea corylifolia L. (PC). Although other constituents of PC have been widely investigated, there are no studies on the biological properties of IDA. In this study, we focused on the anti-inflammatory effects of IDA and evaluated its effects on lipopolysaccharide (LPS)-stimulated macrophages. The results showed that IDA suppressed the production of inflammatory mediators (nitric oxide [NO] and prostaglandin E2 [PGE2]) and proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin-6 [IL-6], and interleukin-1ß [IL-1ß]) without cytotoxicity. In addition, it downregulated the mRNA levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) within the treatment concentrations. In our mechanistic studies, IDA inhibited the phosphorylation of the c-Jun N-terminal kinase (JNK), mitogen-activated protein kinase (MAPK), and protected the nuclear factor of the kappa light polypeptide gene enhancer in the B-cells' inhibitor, alpha (IκB-α), from degradation, thus preventing the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells' (NF-κB) transcription factor. Our results suggest that IDA is a promising compound for attenuating excessive inflammatory responses.
ABSTRACT
Resveratrol is a phytoalexin with multiple bioactive properties, including antioxidative, neuroprotective, cardioprotective, and anticancer effects. However, resveratrol exhibits structural instability in response to UV irradiation, alkaline pH, and oxygen exposure. Thus, resveratrol derivatives have attracted considerable research interest. In this study, we aimed to evaluate the anti-adipogenic effects of pinostilbene hydrate (PH), a methylated resveratrol derivative, in 3T3-L1 cells. We also evaluated the mechanisms underlying the effects of PH on adipogenesis in 3T3-L1 adipocytes. Oil Red O staining, lipid accumulation assay, and triglyceride (TG) content assay revealed that PH significantly inhibited lipid and TG accumulation without cytotoxicity. In addition, we determined that PH decreased the expression of adipogenesis-related transcription factors, such as PPARγ, C/EBPα, SREBP-1c, and FABP4, and the phosphorylation of MAPK and protein kinase B (AKT). Moreover, PH attenuated the expression of CREB and C/EBPß, while increasing the phosphorylation of AMPK and ACC, and decreasing the expression of fatty acid synthase and FABP4. Based on these results, we suggest that PH suppresses adipogenesis in 3T3-L1 cells via the activation of the AMPK signaling pathway and the inhibition of the MAPK and AKT insulin-dependent signaling pathways.
Subject(s)
Adipocytes/metabolism , Adipogenesis/drug effects , MAP Kinase Signaling System/drug effects , Stilbenes/pharmacology , 3T3-L1 Cells , AMP-Activated Protein Kinases/metabolism , Animals , Extracellular Signal-Regulated MAP Kinases/metabolism , Mice , Phosphorylation/drug effects , Transcription Factors/metabolismABSTRACT
Persistent inflammatory reactions promote mucosal damage and cause dysfunction, such as pain, swelling, seizures, and fever. Therefore, in this study, in order to explore the anti-inflammatory effect of 6-methylcoumarin (6-MC) and suggest its availability, macrophages were stimulated with lipopolysaccharide (LPS) to conduct an in vitro experiment. The effects of 6-MC on the production and levels of pro-inflammatory cytokines (interleukin (IL)-1ß, IL-6, tumor necrosis factor (TNF)-α) and inflammatory mediators (nitric oxide (NO), prostaglandin E2 (PGE2)) in LPS-stimulated RAW 264.7 cells were examined. The results showed that 6-MC reduced the levels of NO and PGE2 without being cytotoxic. In addition, it was demonstrated that the increase in the expression of pro-inflammatory cytokines caused by LPS stimulation, was decreased in a concentration-dependent manner with 6-MC treatment. Moreover, Western blot results showed that the protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), which increased with LPS treatment, were decreased by 6-MC treatment. Mechanistic studies revealed that 6-MC reduced the phosphorylation of the mitogen-activated protein kinase (MAPK) family and IκBα in the MAPK and nuclear factor-kappa B (NF-κB) pathways, respectively. These results suggest that 6-MC is a potential therapeutic agent for inflammatory diseases that inhibits inflammation via the MAPK and NF-κB pathways.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Coumarins/pharmacology , Macrophages/drug effects , Mitogen-Activated Protein Kinases/antagonists & inhibitors , NF-kappa B/antagonists & inhibitors , Animals , Anti-Inflammatory Agents, Non-Steroidal/chemistry , Cell Line, Tumor , Coumarins/chemistry , Lipopolysaccharides/antagonists & inhibitors , Lipopolysaccharides/pharmacology , Macrophages/metabolism , Mice , Mitogen-Activated Protein Kinases/metabolism , Molecular Structure , NF-kappa B/metabolism , Nitric Oxide/antagonists & inhibitors , Nitric Oxide/biosynthesis , Phosphorylation/drug effects , RAW 264.7 Cells , Signal Transduction/drug effectsABSTRACT
Nojirimycin (NJ) is a compound in which the oxygen of the ring is replaced with an NH group in the D-glucose structure. NJ, which has a structure similar to D-glucose, is a powerful glucosidase inhibitor and an interesting compound. However, no anti-inflammatory effects of NJ have been reported. Therefore, to investigate its anti-inflammatory effect, the production and expression of inflammatory cytokines, as well as inflammatory mediators, such as iNOS and COX-2, were measured in LPS-stimulated RAW264.7 macrophages. In addition, the effects on the representative inflammatory signaling pathways, the suppression of NF-κ B, and the activation of MAPK were studied. The production of iNOS, COX-2, and inflammatory cytokines (PGE2, IL-6, IL-1ß, and TNF-α) after NJ treatment was significantly inhibited. In addition, NJ showed anti-inflammatory effects through suppression of LPS-induced NF-κ B activation. D-Glucose is structurally similar to NJ. The effects of these substances on RAW264.7 macrophages were evaluated. NJ reduced nitric oxide (NO) levels, whereas D-glucose had no significant effect. Overall, the results suggested that NJ is a potential anti-inflammatory compound.
Subject(s)
1-Deoxynojirimycin/analogs & derivatives , Anti-Inflammatory Agents/pharmacology , Inflammation/drug therapy , NF-kappa B/metabolism , 1-Deoxynojirimycin/pharmacology , Animals , Cell Survival/drug effects , Cyclooxygenase 2/metabolism , Cytokines/drug effects , Cytokines/metabolism , Inflammation Mediators/metabolism , Lipopolysaccharides/pharmacology , MAP Kinase Signaling System/drug effects , Mice , Molecular Structure , Nitric Oxide/biosynthesis , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II , RAW 264.7 Cells , Signal Transduction/drug effectsABSTRACT
Melanin protects our skin from harmful ultraviolet (UV) radiation. However, when produced in excess, it can cause hyperpigmentation disorders, such as melanoma, freckles, lentigo, and blotches. In this study, we investigated the effects of pinostilbene hydrate (PH) on melanogenesis. We also examined the underlying mechanisms of PH on melanin production in B16F10 cells. Our findings indicated that PH significantly inhibits melanin content and cellular tyrosinase activity in cells without causing cytotoxicity. In addition, Western blot analysis showed that PH downregulated the protein levels of microphthalmia-associated transcription factor (MITF), tyrosinase, and other melanogenic enzymes, such as tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2). Although PH activated the phosphorylation of extracellular signal-regulated kinase (ERK), it inhibited p38 mitogen-activated protein kinases (p38). Furthermore, the inhibition of tyrosinase activity by PH was attenuated by treatment with PD98059 (a specific ERK inhibitor). Additionally, p-AKT was upregulated by PH treatment. Finally, the inhibitory effects of PH on melanin content and tyrosinase activity were confirmed in normal human melanocytes. These results suggest PH downregulates melanogenesis via the inhibition of MITF expression, followed by the MAPKase signaling pathways. Thus, PH may be used to treat or prevent hyperpigmentation disorders and in functional cosmetic agents for skin whitening.
Subject(s)
Melanoma/metabolism , Stilbenes/pharmacology , Animals , Cell Line, Tumor , Extracellular Signal-Regulated MAP Kinases/metabolism , Flavonoids/pharmacology , MAP Kinase Signaling System/drug effects , Melanins/metabolism , Melanocytes/drug effects , Melanocytes/metabolism , Melanoma/drug therapy , Mice , Microphthalmia-Associated Transcription Factor/metabolism , Monophenol Monooxygenase/metabolism , Phosphorylation/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/drug effects , Stilbenes/metabolismABSTRACT
Fosfomycin disodium salt (FDS), which is a water-soluble extract, is a bactericidal drug used to inhibit the synthesis of cells. Moreover, it has been found to be effective in the treatment of urinary tract infections. The present study was conducted to investigate the melanogenesis-stimulating effect of FDS in B16F10 cells. Several experiments were performed on B16F10 cells: the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, the melanin content assay, the cellular tyrosinase activity assay, and Western blotting. FDS upregulated the activity of tyrosinase in a dose-dependent manner at a wide concentration range of 0-1 mg/mL, which showed no cytotoxicity. It also increased the melanin content and the activity of the microphthalmia-associated transcription factor (MITF), tyrosinase related protein 1 (TRP-1), and tyrosinase related protein 2 (TRP-2) enzymes in a dose-dependent manner. Western blotting results showed that FDS clearly upregulated the phosphorylation of c-Jun N-terminal kinases (JNK) and p38 pathways. These data are clear evidence of the melanogenesis-inducing effect of FDS in B16F10 murine melanoma cells.
ABSTRACT
Background: Melanin in the skin is the defense against the harmful UV radiation, which is considered as one of the major risk factors for skin cancer. The compound 7,8-dimethoxycoumarin (DMC, C11H10O4), a natural coumarin molecule present in several medicinal plants, possesses antioxidant and anti-inflammatory activities. However, the mechanism underlying its effects on melanogenesis in melanocytes is unclear. Therefore, we investigated the effect of DMC on melanogenesis activation in B16F10 melanoma cells. Methods: We examined the cytotoxic range of DMC on B16F10 melanoma cells and increased effects of melanogenesis, and intracellular tyrosinase activity. In addition, regulation mechanisms were assessed by Western blot analysis. Results: The results showed that DMC significantly increased melanin content and tyrosinase activity in the cells without being cytotoxic. Furthermore, DMC stimulated the expression of tyrosinase, TRP-1, TRP-2, and MITF thereby activating melanin production and Akt phosphorylation was increased in the Akt signaling pathway. on the contrary, interfering with the phosphorylation of ERK in the MAPKs pathway. Conclusions: These results suggest that DMC may serve as a candidate for potential melanin-producing activator and anti-gray hair applications.
Subject(s)
Coumarins/pharmacology , Melanins/biosynthesis , Microphthalmia-Associated Transcription Factor/metabolism , Mitogen-Activated Protein Kinase Kinases/metabolism , Animals , Cell Line, Tumor , Cell Survival , Extracellular Signal-Regulated MAP Kinases/metabolism , Intramolecular Oxidoreductases/metabolism , Melanocytes/drug effects , Melanocytes/metabolism , Melanoma, Experimental , Membrane Glycoproteins/metabolism , Mice , Monophenol Monooxygenase/metabolism , Oxidoreductases/metabolismABSTRACT
Biorenovation is a microbial enzyme-catalyzed structural modification of organic compounds with the potential benefits of reduced toxicity and improved biological properties relative to their precursor compounds. In this study, we synthesized a novel compound verified as formononetin 7-O-phosphate (FMP) from formononetin (FM) using microbial biotransformation. We further compared the anti-inflammatory properties of FMP to FM in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells. We observed that cell viabilities and inhibitory effects on LPS-induced nitric oxide (NO) production were greater in FMP-treated RAW 264.7 cells than in their FM-treated counterparts. In addition, FMP treatment suppressed the production of proinflammatory cytokines such as prostaglandin-E2 (PGE2), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) in a dose-dependent manner and concomitantly decreased the mRNA expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). We also found that FMP exerted its anti-inflammatory effects through the downregulation of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and nuclear factor kappa B (NF-κB) signaling pathways. In conclusion, we generated a novel anti-inflammatory compound using biorenovation and demonstrated its efficacy in cell-based in vitro assays.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Isoflavones/pharmacology , Lipopolysaccharides/pharmacology , Macrophages/metabolism , Animals , Anti-Inflammatory Agents/chemistry , Cell Survival/drug effects , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolism , Cytokines/metabolism , Dinoprostone/metabolism , Inflammation Mediators/metabolism , Isoflavones/chemistry , MAP Kinase Signaling System , Macrophages/drug effects , Mice , NF-kappa B/metabolism , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , RAW 264.7 Cells , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
Tobramycin is an aminoglycoside-based natural antibiotic derived from Streptomyces tenebrarius, which is primarily used for Gram-negative bacterial infection treatment. Although tobramycin has been utilized in clinical practice for a long time, it has exhibited several side effects, leading to the introduction of more effective antibiotics. Therefore, we conducted our experiments focusing on new possibilities for the clinical use of tobramycin. How tobramycin affects skin melanin formation is unknown. This study used B16F10 melanoma cells to assess the effect of tobramycin on melanin production. After cytotoxicity was assessed by MTT assay, melanin content and tyrosinase activity analyses revealed that tobramycin induces melanin synthesis in B16F10 cells. Next, Western blot analyses were performed to elucidate the mechanism by which tobramycin increases melanin production; phosphorylated p38 protein expression was upregulated. Protein inhibitors have been used to elucidate the mechanism of tobramycin. Kanamycin A and B are structurally similar to tobramycin, and 2-DOS represents the central structure of these antibiotics. The effects of these substances on melanogenesis were evaluated. Kanamycin A reduced melanin production, whereas kanamycin B and 2-DOS had no effect. Overall, our data indicated that tobramycin increases melanin production by promoting p38 protein phosphorylation in B16F10 melanoma cells.
ABSTRACT
8-Methoxycoumarin (8-methoxy-chromen-2-one), isolated from R. graveolens L., is able to alleviate arthritis by inhibition of proinflammatory cytokines. However, its effects on melanogenesis have largely remained unreported. The present study examined the effects of 8-methoxycoumarin on melanogenesis in B16F10 murine cells, together with its effect on the mechanism of melanin synthesis. The cells were treated with different concentrations of 8-methoxycoumarin; α-MSH was used as the positive control. We found 8-methoxycoumarin to significantly increase the melanin content of the cells without exerting any cytotoxicity. In addition, it significantly upregulated the expression of tyrosinase and tyrosinase-related protein-1 and 2 via inducing the expression of microphthalmia-associated transcription factor. Furthermore, we demonstrated the involvement of mitogen-activated protein kinase (MAPK) pathway-mediated phosphorylation of p38 and c-Jun N-terminal kinase (JNK), and inhibition of phosphorylation of extracellular signal-regulated kinase (ERK) to be responsible for enhanced melanin production. Use of SB203580 (p38 inhibitor) and SP600125 (p-JNK inhibitor) corroborated these findings. Additionally, we investigated the effects of 8-methoxycoumarin on protein kinase B (AKT) phosphorylation and protein kinase A (PKA) signaling pathway (using H89, a PKA inhibitor). These results suggested that 8-methoxycoumarin increases melanogenesis via the MAPK signaling pathway. Based on these findings, we conclude that 8-methoxycoumarin could serve as a potential compound for treating hypopigmentation disorders. It could also serve as a promising chemical for hair depigmentation treatment in the cosmetic industry.
Subject(s)
Coumarins/pharmacology , MAP Kinase Signaling System/drug effects , Melanins/biosynthesis , Signal Transduction/drug effects , Animals , Anthracenes/pharmacology , Coumarins/administration & dosage , Coumarins/isolation & purification , Cyclic AMP-Dependent Protein Kinases/metabolism , Dose-Response Relationship, Drug , Imidazoles/pharmacology , Melanoma, Experimental/metabolism , Mice , Proto-Oncogene Proteins c-akt/metabolism , Pyridines/pharmacology , Ruta/chemistryABSTRACT
Decreased AMPK-eNOS bioavailability mediates the development of diabetic peripheral neuropathy (DPN) through increased apoptosis and decreased autophagy activity in relation to oxidative stress. Schwann cells are responsible for maintaining structural and functional integrity of neurons and for repairing damaged nerves. We evaluated the neuro-protective effect of cinacalcet on DPN by activating the AMPK-eNOS pathway using db/db mice and human Schwann cells (HSCs). Sciatic nerve of db/db mice was characterized by disorganized myelin, axonal shrinkage, and degeneration that were accompanied by marked fibrosis, inflammation, and apoptosis. These phenotypical alterations were significantly improved by cinacalcet treatment along with improvement in sensorimotor functional parameters. Cinacalcet demonstrated favorable effects through increased expression and activation of calcium-sensing receptor (CaSR)-CaMKKß and phosphorylation of AMPK-eNOS signaling in diabetic sciatic nerve. Cinacalcet decreased apoptosis and increased autophagy activity in relation to decreased oxidative stress in HSCs cultured in high-glucose medium as well. This was accompanied by increased expression of the CaSR, intracellular Ca++ ([Ca++]i) levels, and CaMKKß-LKB1-AMPK signaling pathway, resulting in the net effect of increased eNOS phosphorylation, NOx concentration, Bcl-2/Bax ratio, beclin 1, and LC3-II/LC3-I ratio. These results demonstrated that cinacalcet treatment ameliorates inflammation, apoptosis, and autophagy through increased expression of the CaSR, [Ca++]i levels and subsequent activation of CaMKKß-LKB-1-AMPK-eNOS pathway in the sciatic nerve and HSCs under diabetic condition. Therefore, cinacalcet may play an important role in the restoration and amelioration of DPN by ameliorating apoptosis and improving autophagy.
Subject(s)
Cinacalcet/pharmacology , Diabetic Neuropathies/drug therapy , Nerve Degeneration/drug therapy , Peripheral Nervous System Diseases/drug therapy , Sciatic Nerve/drug effects , AMP-Activated Protein Kinase Kinases , Animals , Apoptosis/drug effects , Autophagy/drug effects , Calcium-Calmodulin-Dependent Protein Kinase Kinase/genetics , Diabetic Neuropathies/genetics , Diabetic Neuropathies/pathology , Gene Expression Regulation/drug effects , Humans , Mice , Mice, Inbred NOD , Nerve Degeneration/genetics , Nerve Degeneration/pathology , Nitric Oxide Synthase Type III/genetics , Oxidative Stress/drug effects , Peripheral Nervous System Diseases/genetics , Peripheral Nervous System Diseases/pathology , Protein Kinases/genetics , Protein Serine-Threonine Kinases/genetics , Schwann Cells/drug effects , Schwann Cells/pathology , Sciatic Nerve/pathology , Signal Transduction/drug effectsABSTRACT
This study was carried out to investigate the antimelanogenic effects of a Polygonum tinctorium flower extract obtained using red nuruk, a traditional Jeju barley-based fermentation starter. We also studied the mechanism of action of the P. tinctorium fermented flower extract (PTFFE) in mouse melanoma cells (B16F10). Cells were treated with various concentrations (62.5, 125 and 250 µg/mL) of PTFFE and the results showed that PTFFE significantly decreased the melanin content and tyrosinase activity without being cytotoxic. In addition, PTFFE strongly inhibited the expression of tyrosinase and tyrosinase-related protein 2 by decreasing the expression of the microphthalmia-associated transcription factor, as shown by a western blot assay. Furthermore, PTFFE inhibited melanogenesis via upregulation of the phosphorylation of extracellular signal-regulated kinase (ERK) and protein kinase B, also known as AKT. We also used inhibitors such as PD98059 (a specific ERK inhibitor) or LY294002 (an AKT inhibitor) to determine whether the signaling pathways are involved. High-performance liquid chromatography fingerprinting showed the presence of a quercetin glucoside (isoquercitrin) and quercetin in PTFFE. To test the potential for PTFFE application as a cosmetic material, we also performed a primary skin irritation test on human skin. In this assay, PTFFE did not induce any adverse reactions at the treatment dose. Based on these results, we suggest that PTFFE may be considered a potential antimelanogenesis candidate for topical applications.
Subject(s)
Extracellular Signal-Regulated MAP Kinases/metabolism , Flowers/chemistry , Gene Expression Regulation/drug effects , Melanoma, Experimental/drug therapy , Plant Extracts/pharmacology , Polygonum/chemistry , Proto-Oncogene Proteins c-akt/metabolism , Animals , Fermentation , Humans , Medicine, Traditional , Melanins/metabolism , Melanoma, Experimental/metabolism , Melanoma, Experimental/pathology , Mice , Monophenol Monooxygenase/metabolism , Phosphorylation , Signal Transduction , Skin/drug effects , Skin Irritancy Tests , Tumor Cells, CulturedABSTRACT
Tyrosinase is the rate-limiting enzyme critical for melanin synthesis. It controls pigmentation in the skin. Activation of tyrosinase is currently the most common approach in the development of tanning and haircare products. Pratol is a 7-hydroxy-4-methoxyflavone found in Trifoliumpratense. In this study, we investigated the effects of pratol on melanogenesis. We also studied the mechanism of action of pratol in B16F10 mouse melanoma cells. The cells were treated with various concentrations (6.25, 12.5, 25, and 50 µM) of pratol to observe its effects. The results showed that pratol significantly increased melanin content and tyrosinase activity in the cells without being cytotoxic. In addition, pratol strongly increased the expression of tyrosinase and tyrosinase-related protein-1 and 2 by enhancing the expression of microphthalmia-associated transcription factor. Furthermore, pratol stimulated melanogenesis via the phosphorylation of p38, c-Jun N-terminal kinases (JNK), and extracellular signal-regulated kinase (ERK). The findings from an assay searching for the inhibitor revealed that SB203580 (a specific p38 inhibitor) or SP600125 (a p-JNK inhibitor) attenuated pratol-induced cellular tyrosinase activity whereas PD98059 (an ERK inhibitor) did not. Additionally, pratol interfered with the phosphorylation of p-AKT. We also found that pratol-induced melanogenesis was reversed by H89, which is a specific protein kinase A inhibitor. The results suggest that, owing to its multi-functional properties, pratol may be a potential tanning agent or a therapeutic agent for hair depigmentation in the cosmetic industry.
Subject(s)
Flavones/pharmacology , Animals , Anthracenes/chemistry , Anthracenes/pharmacology , Cell Line, Tumor , Flavones/chemistry , Imidazoles/chemistry , Imidazoles/pharmacology , Intramolecular Oxidoreductases/metabolism , Mice , Molecular Structure , Oxidoreductases/metabolism , Phosphorylation/drug effects , Proto-Oncogene Proteins c-jun/metabolism , Pyridines/chemistry , Pyridines/pharmacology , Trifolium/chemistryABSTRACT
OBJECTIVE: Electroacupuncture (EA) is a traditional medicine in patients with post-stroke rehabilitation. Brain-derived neurotrophic factor (BDNF) is a potent growth factor involved in recovery following cerebral injury. The aim of the present study was to investigate whether EA increases BDNF levels and facilitates functional recovery. METHODS: Occlusion of the middle cerebral artery was performed in rats (N=12) followed by reperfusion. EA was applied at the GV20 (Baihui) acupoint. Motor and sensory functions were monitored on the Garcia scale for 2 weeks. Expressions of BDNF and receptor tyrosine kinase B (trkB) were determined by immunoblotting and immunohistochemistry. RESULTS: Improvement of Garcia scores, particularly in motor performance, were noted in the group with EA stimulation (p<0.05). With EA application, BDNF was elevated in the ischaemic hemisphere with increased numbers of BDNF(+) cells. Increased expression of trkB was also detected. CONCLUSION: These results indicate that EA at GV20 improves motor recovery and stimulates BDNF/trkB expression in rats with cerebral ischaemia.