• Journal of Life Science
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• v.21 no.9
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• pp.1266-1273
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• 2011

Saponin is a main component of ginseng widely known as an oriental traditional medicinal ingredient. A variety of biological effects of saponin has been reported, but its action related to skin regeneration has remained unclear so far. In this study, the effect of saponin on matrix metalloproteinase as well as its antioxidant effect in cell free system was examined in human dermal fibroblasts. First of all, as a result of investigating the effect of saponin on cell viability using MTT assay, it was shown to increase cell viability below 10 ${\mu}g$/ml, but it also showed cytotoxicity above 25 ${\mu}g$/ml. The antioxidant effect of saponin was exerted by inhibition of $H_2O_2$ in addition to reducing power above 1 ${\mu}g$/ml. In particular, saponin showed a protective effect on DNA oxidation. Furthermore, it was observed that saponin activates MMP-2 and increases MMP-1 activity in gelatin and casein zymography analyses, respectively, indicating that saponin could have potential a therapeutic agent for anti-aging and skin regeneration.

• Journal of Physiology & Pathology in Korean Medicine
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• v.29 no.3
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• pp.273-280
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• 2015

In this study, we used the mixture made from the Rice bran 45 ㎏, Dendropanax 5 ㎏, the sugar of the 10% of the total weight, and the enzyme of the 0.1% of the total weight. After the mixture were fermented for 90 days under 20 $^{\circ}C$, we measured the cell viability and the inhibition rate of the melanin biosynthesis, the activity of tyrosinase and superoxide dismutase (SOD) in malignant melanoma, B16F10 cells, in order to survey the whitening effect and the mechanism of the effects on the sample. As a result, the samples significantly suppressed the cell viability of B16F10 in more than 500 ${{\mu}g}$/㎖ and significantly inhibited the generation of melanin induced by ${\alpha}$-MSH in more than 1,000 ${{\mu}g}$/㎖. Sample decreased the activity of tyrosinase while increased the activity of SOD in dose dependent manner. Therefore, we considered that the fermentation extract made from a Rice bran and Dendropanax will be able to produce high value-added products, if used as a commercial.

• The Korean journal of internal medicine
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• v.34 no.1
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• pp.146-155
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• 2019

Background/Aims: Indoxyl sulfate (IS) is a uremic toxin and an important causative factor in the progression of chronic kidney disease. Recently, paricalcitol (19-nor-1,25-dihydroxyvitamin D2) was shown to exhibit protective effects in kidney injury. Here, we investigated the effects of paricalcitol treatment on IS-induced renal tubular injury. Methods: The fluorescent dye 2',7'-dichlorofluorescein diacetate was used to measure intracellular reactive oxygen species (ROS) following IS administration in human renal proximal tubular epithelial (HK-2) cells. The effects of IS on cell viability were determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays and levels of apoptosis-related proteins (Bcl-2-associated protein X [Bax] and B-cell lymphoma 2 [Bcl-2]), nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) p65, and phosphorylation of mitogen-activated protein kinase (MAPK) and protein kinase B (Akt) were determined by semiquantitative immunoblotting. The promoter activity of $NF-{\kappa}B$ was measured by luciferase assays and apoptosis was determined by f low cytometry of cells stained with f luorescein isothiocyanate-conjugated Annexin V protein. Results: IS treatment increased ROS production, decreased cell viability and induced apoptosis in HK-2 cells. IS treatment increased the expression of apoptosis-related protein Bax, decreased Bcl-2 expression, and activated phosphorylation of MAPK, $NF-{\kappa}B$ p65, and Akt. In contrast, paricalcitol treatment decreased Bax expression, increased Bcl-2 expression, and inhibited phosphorylation of MAPK, $NF-{\kappa}B$ p65, and Akt in HK-2 cells. $NF-{\kappa}B$ promoter activity was increased following IS, administration and was counteracted by pretreatment with paricalcitol. Additionally, flow cytometry analysis revealed that IS-induced apoptosis was attenuated by paricalcitol treatment, which resulted in decreased numbers of fluorescein isothiocyanate-conjugated Annexin V positive cells. Conclusions: Treatment with paricalcitol inhibited IS-induced apoptosis by regulating MAPK, $NF-{\kappa}B$, and Akt signaling pathway in HK-2 cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.5
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• pp.267-274
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• 2019

This study aims to evaluate the effects of Shengmaisan (SMS) and three types of ShengmaisanJiaweifang on the inhibitory effect of melanin synthesis in B16F10 cells, the mechanism of action through tyrosinase, and the antioxidant effect through superoxide dismutase (SOD) activity. In this study, we used ShengmaisanJiaweifangs (SMS, SMSRR, SMSAD, SMSAR) to research the whitening effects in B16F10 cell lines. Shengmaisan (SMS) was a herbal medicine composed of Ginseng Radix, Liriopis Tuber, and Schisandrae Fructus. ShengmaisanJiaweifangs included SMSRR (SMS added with Rehmanniae Radix), SMSAD (SMS added with Asparagi Radix) and SMSAR (SMS added with Astragali Radix). We measured the cell viability, the inhibition rate of the melanin biosynthesis, and the activity of tyrosinase and SOD in malignant melanoma, B16F10 cells, to survey the whitening effect and the mechanism of the impact on the sample. As a result, SMSRR significantly suppressed the cell viability of B16F10 at more than $500{\mu}g/m{\ell}$ and significantly inhibited the generation of melanin induced by ${\alpha}$-MSH at more than $250{\mu}g/m{\ell}$. SMSRR ($500{\mu}g/m{\ell}$) decreased the activity of tyrosinase while increased the activity of SOD. Therefore, we considered that the SMSRR would be able to produce high value-added products more SMS if used as a commercial.

• Physical Therapy Rehabilitation Science
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• v.3 no.1
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• pp.33-37
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• 2014

Objective: Low intensity pulsed ultrasound (LIPUS) has been shown to accelerate cell proliferation and tissue healing in both animal models and clinical trials. However, details of the clinical effects of LIPUS have not been well characterized. The aim of this study was to investigate the effect of LIPUS on mitogen-activated protein kinase (MAPK) activation in rat articular chondrocytes. Design: Cross-sectional study. Methods: Chondrocyte were cultured in six well cell culture plates for 72 hours at $37^{\circ}C$ with 5% $CO_2$, and then exposed to LIPUS at 1.5 MHz frequency and $30-mW/cm^2$ power. Changes in chondrocyte activities were evaluated in response to oxydative stress in dose-dependent (0 and 300 uM) and time-dependent (0-24 hr) manner. The cell viability were analyzed using MTT [3-(4.5-dimethylthiazol-2-yl)-2.5 diphenyltetrazolium bromide]. The expression of p38 MAPK was measured using western blotting. Results: Oxidative stress was induced in rat chondrocytes using hydrogen peroxide ($H_2O_2$). The cell viability was decreased in chondrocytes after the $H_2O_2$ dose and time-dependent treatment. The p38 MAPK phosphorylation occurred at a significantly increased rate after $H_2O_2$ treated (p<0.05). Expression of p38 MAPK was decreased in the p38 inhibitor groups compared with the oxidative stress-induced chondrocyte damage via the p38 MAPK signaling pathways (p<0.05). Conclusions: It could be concluded that LIPUS can inhibit oxidative stress-induced chondrocyte damage via the p38 MAPK signaling pathways.

• Journal of the Korean Applied Science and Technology
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• v.36 no.1
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• pp.258-268
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• 2019

The extracts of AS contain in alloxanthin, halocynthiaxanthin, astaxanthin and 13 kinds of carotenoids. The aim of the study was to assess the anti-oxidant activity and cell viability of AS. The anti-oxidant activity was determined by using DPPH radical inhibition activity and superoxide dismutase (SOD)-like activity. The results of cell viability assay showed that the extracts from AS were cytotoxic at concentrations above $5.0mg/m{\ell}$. This study was designed to examine inflammation induced by LPS, protection effect by UVB and the toxicity of Ascidian shell extract(ASE) as a functional cosmetic ingredient. Evaluation of embryo toxicity resulted in embryo coagulation and mortality when treated at 5.0, 10.0, $20.0mg/m{\ell}$. At the lowest concentration of $1.0mg/m{\ell}$, hatchability resulted in 100.0 % rate. The results of arrhythmia measurement in larvae showed similarity to the evaluation of embryo toxicity. This result demonstrated that toxicity is present at concentrations greater than $5.0mg/m{\ell}$. The protective effect of ASE on LPS and UVB-induced in the zebrafish was investigated. Intracellular reactive oxygen species(ROS) generated by the exposure of zebrafish to LPS, UVB-radiation were significantly decreased after treatment with ASE at $0.1mg/m{\ell}$. As a result, ASE similarly reduced UVB-induced ROS generation and cell death in live zebrafsih. Therefore, it is suggested that ASE has anti-Inflammatory effects and can possibly be used as a functional substance for skin protection in the future.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.32 no.2
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• pp.11-23
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• 2019

Objectives : The purpose of this study is to investigate the anti-inflammatory effect of Cheongsimyanggyeoksan(CYS) water extract in vitro and in vivo. Methods : To evaluate the anti-inflammatory effect of CYS, Raw 264.7 cells were pretreated with $3-300{\mu}g/m{\ell}$ of CYS for 1h, and then exposed to $1{\mu}g/m{\ell}$ of LPS. The cell viability was detected by MTT assay. Productions of nitric oxide(NO) and pro-inflammatory cytokines were measured in culture media. Protein levels of inducible nitric oxide synthase(iNOS) and Nuclear factor-${\kappa}$B($NF-{\kappa}B$) were determined by immunoblot analysis. The effect of CYS on acute inflammation in vivo was evaluated thorugh measurment of carrageenan-induced paw edema. Results : In vitro study, cell viability assay CYS treatment of $3-300{\mu}g/m{\ell}$ has no cytotoxicity in Raw 264.7 cells. LPS-induced NO production was significantly inhibited by pretreatment with $30-300{\mu}g/m{\ell}$ of CYS. Production of interleukin-6, -$1{\beta}$ and tumor necrosis factor-${\alpha}$ by LPS were significantly decreased by CYS pretreatment. CYS reduced LPS-mediated iNOS expression. Moreover, CYS significantly induced $I-{\kappa}B{\alpha}$ expression and reduced $NF-{\kappa}B$ expression. In vivo study, CYS significantly reduced the increases of paw swelling. Conclusions : These results suggest the clinical basis of CYS for the treatment of inflammatory diseases.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.8-8
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• 2019

The stringent response (SR) is characterized as a bacterial defense mechanism in response to various growth-inhibiting stresses. It is activated by accumulation of a small nucleotide regulator, (p)ppGpp, and induces global changes in bacterial transcription and translation. Recent work from our group has shown that (p)ppGpp plays a critical role in virulence and survival in Vibrio cholerae. The genes, relA and relV, are involved in the production of (p)ppGpp, while the spoT gene encodes an enzyme that hydrolyzes it in V. cholerae. A mutant strain defective in (p)ppGpp production (i.e. ${\Delta}relA{\Delta}relV{\Delta}spoT$ mutant) lost the ability to produce cholera toxin (CT) and lost their viability due to uncontrolled production of organic acids, when grown with extra glucose. In contrast, the ${\Delta}relA{\Delta}spoT$ mutant, a (p)ppGpp overproducer strain, produced enhanced level of CT and exhibited better growth in glucose supplemented media via glucose metabolic switch from organic fermentation to acetoin, a neutral fermentation end product, fermentation. These findings indicates that (p)ppGpp, in addition to its well-known role as a SR mediator, positively regulates CT production and maintenance of growth fitness in V. cholerae. This implicates SR as a promising drug target, inhibition of which may possibly downregulate V. cholerae virulence and survival fitness. Therefore, we screened a chemical library and identified a compound that induces medium acidification (termed iMAC) and thereby loss of wild type V. cholerae viability under glucose-rich conditions. Further, we present a potential mechanism by which the compound inhibits (p)ppGpp accumulation. Together, these results indicate that iMAC treatment causes V. cholerae cells to produce significantly less (p)ppGpp, an important regulator of the bacterial virulence and survival response, and further suggesting that it has a therapeutic potential to be developed as a novel antibacterial agent against cholera.

• The Korean Journal of Pain
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• v.34 no.1
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• pp.19-26
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• 2021

Background: Prolotherapy is a proliferation therapy as an alternative medicine. A combination of dextrose solution and lidocaine is usually used in prolotherapy. The concentrations of dextrose and lidocaine used in the clinical field are very high (dextrose 10%-25%, lidocaine 0.075%-1%). Several studies show about 1% dextrose and more than 0.2% lidocaine induced cell death in various cell types. We investigated the effects of low concentrations of dextrose and lidocaine in fibroblasts and suggest the optimal range of concentrations of dextrose and lidocaine in prolotherapy. Methods: Various concentrations of dextrose and lidocaine were treated in NIH-3T3. Viability was examined with trypan blue exclusion assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Migration assay was performed for measuring the motile activity. Extracellular signal-regulated kinase (Erk) activation and protein expression of collagen I and α-smooth muscle actin (α-SMA) were determined with western blot analysis. Results: The cell viability was decreased in concentrations of more than 5% dextrose and 0.1% lidocaine. However, in the concentrations 1% dextrose (D1) and 0.01% lidocaine (L0.01), fibroblasts proliferated mildly. The ability of migration in fibroblast was increased in the D1, L0.01, and D1 + L0.01 groups sequentially. D1 and L0.01 increased Erk activation and the expression of collagen I and α-SMA and D1 + L0.01 further increased. The inhibition of Erk activation suppressed fibroblast proliferation and the synthesis of collagen I. Conclusions: D1, L0.01, and the combination of D1 and L0.01 induced fibroblast proliferation and increased collagen I synthesis via Erk activation.

• CELLMED
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• v.11 no.3
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• pp.16.1-16.7
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• 2021

Cancer is an abnormal growth of cells in body which leads to death. These cells are born due to imbalance in cell proliferation mechanism. In 2018, WHO released new statistics on cancer incidence, mortality, and prevalence worldwide i.e., GLOBOCAN 2018 estimates for 28 types of cancer in which more prevalence of cervix and breast cancer. According to survey, in India about 7.8 million cancer deaths and 11.5 million new cases arise in 2018, which will increase to 19.3 million new cases per year by 2025. Though breast cancer as such is not explained anywhere in Ayurvedic compendia, correlations can be done with the Stana Arbuda. Ayurveda, the ancient system of medicine came into existence 1000's of years ago with an objective of maintaining the health of people and treating diseases. Many herbs used in Ayurveda have been screened for activity against cancer and in-vitro and in-vivo studies have given promising leads. The plant, called as "Mother of Medicine", Haritaki has been extensively studied for its various ailments because of its extraordinary healing potency. Haritaki (Terminalia chebula Retz.), Family: Combretaceae have a great therapeutic value and is widely distributed in India. Dried fruit of Terminalia chebula contains high quantities phenolic compounds consist of ellagic acid, gallic acid and chebulic acid. The fruit extract of T. chebula is having different biological properties like anticancer, antioxidant, hepatic and renal protective activities etc. In this study, we focus on the use of CO₂ extract of Terminalia chebula, on the breast cancer cell line MDA-MB-231. All tests proved that CO₂ extract of Terminalia chebula containing active chemical component, therefore our experiment showed the positive results for CO₂ extract of Terminalia chebula against breast cancer cell line cancer MDA-MB-231. The MTT assay results were used to evaluate the anti-cancer activity of the extract. The percentage of cell growth and cell viability were calculated from tabulated result values of MTT assay. Cell viability MTT assay also showed significant growth inhibition, at the same time statistical analysis of MTT assay also proved significant results.