• Journal of Ginseng Research
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• v.44 no.2
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• pp.341-349
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• 2020

Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)⁺/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD⁺/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD⁺/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.337-346
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• 2023

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2005.11a
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• pp.38-38
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• 2005

• Korean Journal of Pharmacognosy
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• v.46 no.2
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• pp.154-159
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• 2015

This study was to evaluate the effect of isopropyl alcohol fraction of ultrasonication processed ginseng flower buds(GFB-IF) on the collagen synthesis activity and inhibition of MMP-1 suppression in UV-irradiated human dermal fibroblasts. The higher contents of ginsenoside Rg2(8.234%), Rh1(5.749%), F4(3.881%) in isopropyl alcohol fraction of ginseng flower buds obtained by ultrasonication process at 600W(100℃) for 16 hours. GFB-IF had collagen synthesis effect. GFB-IF induced a significant dose-dependent decrease in the expression for MMP-1 protein. These results suggest that GFB-IF is a potential candidate for the prevention and treatment of wrinkle improving.

• Macromolecular Research
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• v.11 no.5
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• pp.368-374
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• 2003

Porous scaffolds composed of gelatin and polysaccharides such as hyaluronic acid and $\beta$-glucan were prepared by using the freeze-drying method after cross-linking with l-ethyl-(3-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The scaffold had an inter-connected pore structure with the sufficient pore size for use as a support for the growth of fibroblasts. Results for the contact angle and cell attachment confirmed that high gelatin content in a mixture was suitable for cellular attachment and distribution in two- or three-dimensional fibroblast cultures. However, the addition of polysaccharides aroused the synergistic effects of morphologic and mechanical property of gelatin-based scaffolds. To prepare the artificial dermis for the wound dressing to mimic the normal human dermal skin, fibroblasts were isolated from a child's foreskin, and cultured in gelatin-based scaffolds. An in vivo study showed that the artificial dermis containing the fibroblasts enhanced the wound healing rate and re-epithelialization of a full-thickness skin defect rather than the acellular scaffold after one week.

• Korean Journal of Medicinal Crop Science
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• v.22 no.5
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• pp.331-338
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• 2014

Dendrobium moniliforme (DM) is a valuable and versatile herbal medicine with the anecdotal claims of antioxidant and anti-inflammation. In the present study, we investigated the whitening and anti-wrinkling effects of DM under various conditions with B16F10 melanoma cells and human dermal fibroblasts. The DM extract inhibited melanin contents and tyrosinase activity in a dose-dependent manner, compared with untreated group. Treatment of the DM extract effectively suppressed the ${\alpha}$-MSH-stimulated melanin formation, tyrosinase activity and dendrite outgrowth. Moreover, the ${\alpha}$-MSH-induced mRNA expressions of tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2) and protein expression of tyrosinase were significantly attenuated by DM treatment. We also investigated the DM increased the production of type I procollagen and inhibited TNF-${\alpha}$-induced mRNA expressions of MMP-1, -3 in the human dermal fibroblast. These results indicate that DM may be a great cosmeceutical ingredient for its whitening and anti-wrinkle effects.

• Journal of Biomedical Engineering Research
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• v.27 no.5
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• pp.218-223
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• 2006

We examined the effects of ${\beta}$-glucan-reinforced PLGA film and scaffold on HDFs (human dermal fibroblast) attachment and proliferation. The PLGA films were prepared by simple solvent-casting method. The prepared films were grafted with $(1{\to}3)(1{\to}6)-{\beta}-glucan$ in various ratios after plasma treatment on surface. The surface of the film was characterized by contact angle measurement, scanning electron microscope (SEM), and Fourier-transform infrared spectrophotometer (FT-IR). The amount of $(1{\to}3)(1{\to}6)-{\beta}-glucan$ in the prepared film was indirectly determined by phenol-sulfuric acid method. The HDFs (Human dermal fibroblasts) were used to evaluate the cell attachment and proliferation on PLGA specimens before and after plasma/${\beta}-glucan$ treatment. The result showed that the plasma treated groups exhibited more mont of ${\beta}-glucan$ might be grafted than the non plasma treated groups. Cell attachment was significantly enhanced in the plasma/${\beta}-glucan$ grafted group after 4 hours incubation (p<0.05) due to the improved hydrophilicity and cytoactivity effect of the ${\beta}-glucan$. The cell proliferation of plasma/${\beta}-glucan$ (2mg/ml) grafted group was the highest rate among the groups (p<0.05).

• Journal of Ginseng Research
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• v.46 no.5
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• pp.646-656
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• 2022

Background: In addition to its use as a health food, ginseng is used in cosmetics and shampoo because of its extensive health benefits. The ginsenoside, Rh2, is a component of ginseng that inhibits tumor cell proliferation and differentiation, promotes insulin secretion, improves insulin sensitivity, and shows antioxidant effects. Methods: The effects of Rh2 on cell survival, extracellular matrix (ECM) protein expression, and cell differentiation were examined. The antioxidant effects of Rh2 in UV-irradiated normal human dermal fibroblast (NHDF) cells were also examined. The effects of Rh2 on mitochondrial function, morphology, and mitophagy were investigated in UV-irradiated NHDF cells. Results: Rh2 treatment promoted the proliferation of NHDF cells. Additionally, Rh2 increased the expression levels of ECM proteins and growth-associated immediate-early genes in ultraviolet (UV)-irradiated NHDF cells. Rh2 also affected antioxidant protein expression and increased total antioxidant capacity. Furthermore, treatment with Rh2 ameliorated the changes in mitochondrial morphology, induced the recovery of mitochondrial function, and inhibited the initiation of mitophagy in UV-irradiated NHDF cells. Conclusion: Rh2 inhibits mitophagy and reinstates mitochondrial ATP production and membrane potential in NHDF cells damaged by UV exposure, leading to the recovery of ECM, cell proliferation, and antioxidant capacity.

• YAKHAK HOEJI
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• v.48 no.6
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• pp.358-363
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• 2004

Matrix metalloproteinases (MMPs) are known to play an important role in photoaging by mediating the degradation of extracellular matrix proteins. In this study, to develop a n ew anti-aging agent, we investigated the antioxidant activity and the inhibitory effect of Melothria heterophylla extract on expression of MMP-1 in UVA-irradiated human dermal fibroblasts and MMP-1 activity. The M.heterophylla extract was found to scavenge radicals and reactive oxygen species (ROS) with the $SC_{50}$ values of $13{\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and $20{\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. UVA-induced MMP-1 expression was reduced about 80% by $100{\mu}g/ml$ of the M.heterophylla extract but MMP-1 Mrna expression was not inhibited. Therefore, we conclude that the M.heterophylla extract significantly inhibits MMP-1 expression at the protein level. Also, the M.heterophylla extract inhibited MMP-1 activity in a dose dependent manner. From these results, we suggest that the M.heterophylla extract can be used as a new anti-aging agent by antioxidant activity, regulation of UVA-induced MMP-1 production, and inhibition of MMP-1 activity.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2179-2183
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• 2014

Background: In vitro, in vivo and clinical studies have demonstrated anti-cancer effects of deuterium depleted water (DDW). The nature of this agents action, cytotoxic or cytostatic, remains to be elucidated. We here aimed to address the point by examining effects on different cell lines. Materials and Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) -based cytotoxicity analysis was conducted for human breast, stomach, colon, prostate cancer and glioblastoma multiforme cell lines as well as human dermal fibroblasts. The cell lines were treated with decreasing deuterium concentrations of DDW alone, paclitaxel alone and both. One way analysis of variance (ANOVA) was used for statistical analysis. Results: Treatment with different deuterium concentrations of DDW alone did not impose any significant inhibitory effects on growth of cell lines. Paclitaxel significantly decreased the survival fractions of all cell lines. DDW augmented paclitaxel inhibitory effects on breast, prostate, stomach cancer and glioblastoma cell lines, with influence being more pronounced in breast and prostate cases. Conclusions: DDW per se does not appear to have inhibitory effects on the assessed tumor cell lines as well as normal fibroblasts. As an adjuvant, however, DDW augmented inhibitory effects of paclitaxel and thus it could be considered as an adjuvant to conventional anticancer agents in future trials.