• Journal of Life Science
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• v.31 no.11
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• pp.1019-1027
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• 2021

There are a lot of efforts to develop new compounds having skin whitening effect from natural products. Sparassis crispa is a medicinal mushroom containing more than 40% β-glucan, which exhibits anticancer and immunostimulating effects. The aim of this study was to assess the availability of β-glucan extracted from cauliflower mushroom S. crispa as a skin whitener through the evaluation of inhibitory effects of melanin synthesis and tyrosinase activity and their mechanisms. B16F1 cells were treated with S. crispa β-glucan (10, 100, and 1,000 ㎍/ml, respectively) and α-melanocyte stimulating hormone (α-MSH), simultaneously. Content of melanin synthesis and tyrosinase activity were determined. The expressions levels of tyrosinase, tyrosinase related protein-1 (TRP-1), TRP-2 and microphthalmia-associated transcription factor (MITF) were also measured by western blotting. Treatment with 10, 100 and 1,000 ㎍/ml S. crispa β-glucan and 200 nM α-MSH significantly decreased melanin synthesis by 13.9%, 18.7% and 39.5%, respectively, and tyrosinase activity by 15.6%, 26.9% and 43.2%, respectively, compared to the α-MSH alone group. In addition, S. crispa β-glucan inhibited expressions of tyrosinase, TRP-1, TRP-2 and MITF induced by α-MSH. These results indicated that S. crispa β-glucan inhibited MITF expression, thereby reducing tyrosinase expression and inhibiting melanin production in B16F1 melanoma cells. Therefore, S. crispa β-glucan might be available as a skin whitener.

• The Journal of Internal Korean Medicine
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• v.43 no.4
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• pp.514-528
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• 2022

Objective: To examine the effects of Trichosanthes kirilowii Maximowicz extract (TE) and Trichosanthes kirilowii Maxi mowicz Cheonghyeol Plus Phellinus linteus Cheonghyeol plus (TCP) on anti-inflammatory factor expression in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were activated with TNF-α and then treated with TE and TCP. The expression levels were then measured for intracellular genes (KLF2, eNOS, MCP-1, ICAM-1, and VCAM-1), proteins (KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, ERK, and JNK, p38), and extracellular biomarkers (ICAM-1, VCAM-1, and MCP-1). Results: 1. TCP at concentrations of 100 ㎍/mL or greater significantly increased the expression of KLF2 and eNOS intracellular genes and significantly decreased the expression of ICAM-1, VCAM-1, and MCP-1 genes compared to the control group. 2. TCP at concentrations of 100 ㎍/mL or greater significantly increased the expression of KLF2, eNOS proteins compared to the control group, and significantly reduced the expression of VCAM-1, ICAM-1, MCP-1, ERK, and p38 proteins. However, JNK protein phosphorylation showed no significant change compared to the control group. 3. TCP at concentrations of 100 ㎍/mL or more significantly decreased the production of MCP-1, ICAM-1, and VCAM-1 extracellular biomarkers compared to the control group. 4. TE at a concentration of 100 ㎍/mL did not cause any significant change in the expression of intracellular genes or proteins, in the production of the extracellular biomarker MCP-1, or in the amount of JNK protein compared to the control group. Other intracellular genes, proteins, and extracellular biomarker expression showed the same trend as observed with TCP exposure. Conclusion: This study experimentally confirmed that TE and TCP could be effective in preventing or inhibiting various inflammatory vascular diseases due to their anti-inflammatory effects.

• The Korea Journal of Herbology
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• v.29 no.2
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• pp.39-45
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• 2014

Objectives : The purpose of this study was to investigate antiaging and antioxidant effects on cultured human skin fibroblast with 80% ethanol extracts of plants including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Methods : An ethanol extract of three medicinal plants including stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Extracts were assessed to determine the mechanism of antioxidant and antiaging activities. Antioxidant activity of extract was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. These extracts were tested for cell viability on HS68 skin fibroblast by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. We investigated the effects of Ultraviolet-B irradiation on cytotoxicity, type 1 collagen, elastin level and oxidative damage in cultured human skin fibroblast (HS68). Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Results : The extracts obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The extracts of complex herbal medicine showed low cytotoxicity as more than 100% cell viability in 100ppm/ml concentration. HS68 fibroblasts were survived 70% at 120 $mJ/cm^2$ UVB irradiation and treated tumor necrosis factor (TNF)-alpha. The levels of aging factors and cytotoxicity were decreased by ethanol extract of complex herbal medicine. Conclusions : These results suggest that ethanol extracts of complex medicinal plants of including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus may have value as the potential antioxidant and antiaging medicinal plant.

• The Korea Journal of Herbology
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• v.35 no.1
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• pp.57-68
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• 2020

Objective : The aim of present study was to clarify the effect of Areca Semen and Toosendan Fructus Mixture (AT-mix) on chronic reflux esophagitis (CRE) in rats. Methods : The antioxidant activity of AT-mix was measured through DPPH and ABTS radical scavenging activities in vitro. CRE was induced in SD rats (5 weeks, male) by ligating the border forestomach and granular portion with 2-0 silk and the duodenum near the pyloric portion was covered with 2-mm wide piece of 18-Fr Nélaton catheter. And then rats were treated AT-mix 200 mg/kg one daily for 14 days. The anti-oxidant and inflammatory protein levels were evaluated using western blotting. Results : Gross lesion of esophageal mucosa after AT-mix treatment showed a superior enhancement compared with that of CRE control rats. AT-mix treatment strongly reduced both DPPH and ABTS radical scavenging activities (DPPH, IC₅₀ 8.15±0.14 ㎍/mL; ABTS, IC₅₀ 24.69±0.03 ㎍/mL, repspectively). Levels of the NADPH oxidase subunit including NOX4 and p22^phox increased in CRE control rats. Otherwise, AT-mix treatment significantly reduced. The activation of Nuclear factor-erythroid 2-related factor 2 (Nrf2) led to significantly the up-regulation of HO-1. The inhibition of IκBα phosphorylation led to NF-κB inactivation. Subsequently, NF-κB inactivation significantly induced the decrease of COX-2, iNOS, TNF-α, and IL-6 protein expressions. Conclusion : Taken together, these results suggest that AT-mix treatment can attenuate the esophageal mucosal ulcer though inhibiting NF-κB pathway and enhancing Nrf2/HO-1 pathway. Thus, the additional mechanism study about AT-mix would need for the development as a safe herbal therapy for CRE.

• Journal of Veterinary Science
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• v.24 no.6
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• pp.83.1-83.12
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• 2023

Background: Ellipticine (Ellip.) was recently reported to have beneficial effects on the differentiation of adipose-derived stem cells into mature chondrocyte-like cells. On the other hand, no practical results have been derived from the transplantation of bone marrow stem cells (BMSCs) in a rabbit osteoarthritis (OA) model. Objectives: This study examined whether autologous BMSCs incubated with ellipticine (Ellip.+BMSCs) could regenerate articular cartilage in rabbit OA, a model similar to degenerative arthritis in human beings. Methods: A portion of rabbit articular cartilage was surgically removed, and Ellip.+BMSCs were transplanted into the lesion area. After two and four weeks of treatment, the serum levels of proinflammatory cytokines, i.e., tumor necrosis factor α (TNF-α) and prostaglandin E2 (PGE2), were analyzed, while macroscopic and micro-computed tomography (CT) evaluations were conducted to determine the intensity of cartilage degeneration. Furthermore, immuno-blotting was performed to evaluate the mitogen-activated protein kinases, PI3K/Akt, and nuclear factor-κB (NF-κB) signaling in rabbit OA models. Histological staining was used to confirm the change in the pattern of collagen and proteoglycan in the articular cartilage matrix. Results: The transplantation of Ellip.+BMSCs elicited a chondroprotective effect by reducing the inflammatory factors (TNF-α, PGE2) in a time-dependent manner. Macroscopic observations, micro-CT, and histological staining revealed articular cartilage regeneration with the downregulation of matrix-metallo proteinases (MMPs), preventing articular cartilage degradation. Furthermore, histological observations confirmed a significant boost in the production of chondrocytes, collagen, and proteoglycan compared to the control group. Western blotting data revealed the downregulation of the p38, PI3K-Akt, and NF-κB inflammatory pathways to attenuate inflammation. Conclusions: The transplantation of Ellip.+BMSCs normalized the OA condition by boosting the recovery of degenerated articular cartilage and inhibiting the catabolic signaling pathway.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1213-1227
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• 2023

Fetal growth restriction (FGR) is a prevalent obstetric condition. This study aimed to investigate the role of Toll-like receptor 9 (TLR9) in regulating the inflammatory response and gut microbiota structure in FGR. An FGR animal model was established in rats, and ODN1668 and hydroxychloroquine (HCQ) were administered. Changes in gut microbiota structure were assessed using 16S rRNA sequencing, and fecal microbiota transplantation (FMT) was conducted. HTR-8/Svneo cells were treated with ODN1668 and HCQ to evaluate cell growth. Histopathological analysis was performed, and relative factor levels were measured. The results showed that FGR rats exhibited elevated levels of TLR9 and myeloid differentiating primary response gene 88 (MyD88). In vitro experiments demonstrated that TLR9 inhibited trophoblast cell proliferation and invasion. TLR9 upregulated lipopolysaccharide (LPS), LPS-binding protein (LBP), interleukin (IL)-1β and tumor necrosis factor (TNF)-α while downregulating IL-10. TLR9 activated the TARF3-TBK1-IRF3 signaling pathway. In vivo experiments showed HCQ reduced inflammation in FGR rats, and the relative cytokine expression followed a similar trend to that observed in vitro. TLR9 stimulated neutrophil activation. HCQ in FGR rats resulted in changes in the abundance of Eubacterium_coprostanoligenes_group at the family level and the abundance of Eubacterium_coprostanoligenes_group and Bacteroides at the genus level. TLR9 and associated inflammatory factors were correlated with Bacteroides, Prevotella, Streptococcus, and Prevotellaceae_Ga6A1_group. FMT from FGR rats interfered with the therapeutic effects of HCQ. In conclusion, our findings suggest that TLR9 regulates the inflammatory response and gut microbiota structure in FGR, providing new insights into the pathogenesis of FGR and suggesting potential therapeutic interventions.

• Journal of Life Science
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• v.33 no.11
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• pp.859-867
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• 2023

Lupeol is a type of pentacyclic triterpene that has been reported to have therapeutic effects for treating many diseases; however, its effect on insulin resistance is unclear clear. This study examined the inhibitory effect of lupeol on the serine phosphorylation of insulin receptor substrate-1 in insulin resistance-induced 3T3-L1 adipocytes. 3T3-L1 cells were cultured and treated with tumor necrosis factor-α (TNF-α) for 24 hours to induce insulin resistance. Cells treated with different concentrations of lupeol (15 μM or 30 μM) or 100 nM of rosiglitazone were incubated. Then, lysed cells underwent western blotting. Lupeol exhibited a positive effect on the negative regulator of insulin signaling and inflammation-activated protein kinase caused by TNF-α in adipocytes. Lupeol inhibited the activation of protein tyrosine phosphatase-1B (PTP-1B)-a negative regulator of insulin signaling-and c-Jun N-terminal kinase (JNK); it was also an inhibitor of nuclear factor kappa-B kinase (IKK) and inflammation-activated protein kinases. In addition, Lupeol downregulated serine phosphorylation and upregulated tyrosine phosphorylation in insulin receptor substrate-1. Then, the downregulated phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway was activated, the translocation of glucose transporter type 4 was stimulated to the cell membrane, and intracellular glucose uptake increased in the insulin resistance-induced 3T3-L1 adipocytes. Lupeol may improve TNF-α-induced insulin resistance by downregulating the serine phosphorylation of insulin receptor substrate 1 by inhibiting negative regulators of insulin signaling and inflammation-activated protein kinases in 3T3-L1 adipocytes.

• Food Science and Preservation
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• v.30 no.3
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• pp.502-513
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• 2023

This study aimed to evaluate the anti-adipogenic and anti-inflammation effects of extract from Maclura tricuspidata twig fermented with Ganoderma lucidum mycelium (EMFG) in 3T3-L1 preadipocytes. 3T3-L1 adipocytes were treated with 100, 200, 300 ㎍/mL of EMFG. The result showed that EMFG dose-dependently inhibited the accumulation of intracellular lipid content in differentiated 3T3-L1 adipocytes and enhanced increase of adiponectin release and inhibition of leptin release. EMFG treatment reduced expression of adipogenic transcriptional factor such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT-enhancer-binding protein α (C/EBPα). EMFG also decreased production of lipopolysaccharide (LPS)-induced inflammatory cytokine [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1)] and the protein expression of cyclooxygenase-2 (COX-2) and inducible NOS (iNOS) in differentiated 3T3-L1 adipocytes. The study demonstrated that EMFG inhibited adipogenesis and inflammation in a dose-dependent manner. These findings suggest that EMFG may have potential as an anti-obesity and anti-metabolic disease agent that works by inhibiting adipogenesis and inflammation.

• Korean Journal of Clinical Laboratory Science
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• v.55 no.4
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• pp.284-290
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• 2023

Adefovir dipivoxil (ADV) is used for the treatment of hepatitis and acquired immunodeficiency syndrome, but long-term use can cause osteoporosis. In this study, the effect of ADV on the osteocyte maturation process was evaluated at the level of undifferentiated cells using mesenchymal stem cells (MSCs) and osteoblasts (MG63). First, MSCs and MG63 cells were treated with ADV at different concentrations, and then a Cell Counting Kit-8 analysis was performed to determine the effect on the proliferation of each cell. Additionally, crystal violet and Hoechst staining were performed for the morphological analysis of each cell and nucleus. To determine the cause of cell hypertrophy, the transforming growth factor-beta (TGF-β) expression was investigated, and alkaline phosphatase (ALP) staining and activity were measured to determine the degree of differentiation of the MSCs and MG63 cells into mature osteocytes. The results confirmed that the ADV increases the expression of TGF-β in MSCs and MG63 cells, causing cellular and nuclear hypertrophy, and can cause osteoporosis by inhibiting cell proliferation and affecting the differentiation of mature osteocytes. Therefore, it is believed that these results can be used as a basis for understanding the adverse effects of ADV at a cytological level in basic medicine and clinical research.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.46-52
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• 2023

The most abundant flavanone of grapefruits, naringenin (NN), is well known for its hepatoprotective, anti-lipid peroxidation and anti-carcinogenic effects. We generated three derivatives from NN using this technique in previous studies. Among them, it was confirmed that naringenin-7-O-phosphate (N7P), whose biological and physicochemical properties were not reported, showed a water solubility 45 times higher than that of NN. Therefore, in this study, the anti-inflammatory activity was evaluated in RAW 264.7 cells to investigate the potential physiological activity of N7P. As a result, N7P showed nitric oxide (NO) inhibitory activity at concentrations that did not show toxicity. In addition, prostaglandin E₂ (PGE₂) showed significant inhibitory activity from the lowest concentration of 12.5 μM and showed increased inhibitory activity compared to NN. In addition, as a result of western blot, N7P showed increased cyclooxygenase-2 (COX-2) inhibitory activity than NN, and effectively inhibited NO and PGE₂ by significantly inhibiting their expression pathways. N7P also inhibited inflammatory cytokines, including tumor necrosis factor-α, interleukin-6. Based on these results, we propose that N7P can be used as a potent antiinflammatory agent.