• Journal of Microbiology and Biotechnology
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• 제29권9호
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• pp.1349-1360
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• 2019

Chronic exposure to ultraviolet (UV) radiation, regarded as a major cause of extrinsic aging or photoaging characterized by wrinkle formation and skin dehydration, exerts adverse effects on skin by causing the overproduction of reactive oxygen species. Agastache rugosa Kuntze, known as Korean mint, possesses a wide spectrum of biological properties including anti-oxidation, anti-inflammation, and anti-atherosclerosis. Previous studies have reported that A. rugosa protected human keratinocytes against UVB irradiation by restoring the anti-oxidant defense system. However, the anti-photoaging effect of A. rugosa extract (ARE) in animal models has not yet been evaluated. ARE was orally administered to hairless mice at doses of 100 or 250 mg/kg/day along with UVB exposure for 12 weeks. ARE histologically improved UVB-induced wrinkle formation, epidermal thickening, erythema, and hyperpigmentation. In addition, ARE recovered skin moisture by improving skin hydration and transepidermal water loss (TEWL). Along with this, ARE increased hyaluronic acid levels by upregulating HA synthase genes. ARE markedly increased the density of collagen and the amounts of hydroxypoline via two pathways. First, ARE significantly downregulated the mRNA expression of matrix metalloproteinases responsible for collagen degradation by inactivating the mitogen-activated protein kinase/activator protein 1 pathway. Second, ARE stimulated the transforming growth factor beta/Smad signaling, consequently raising the mRNA levels of collagen-related genes. In addition, ARE not only increased the mRNA expression of anti-oxidant enzymes but also decreased inflammatory cytokines by blocking the protein expression of nuclear factor kappa B. Collectively, our findings suggest that A. rugosa may be a potential preventive and therapeutic agent for photoaging.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.1047-1055
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• 2015

For the search of a potent first-in-class compound to inactivate macrophages responsible for inflammatory responses, in the present study, we investigated the anti-nflammatory effects of YH-1118, a novel synthetic compound, in a lipopolysaccharide (LPS)-stimulated mouse macrophage cell line, Raw 264.7. YH-1118 inhibited LPS-induced nitric oxide (NO) production and inducible NO synthase (iNOS) expression at both the protein and mRNA levels. The suppression of LPS-induced iNOS expression by YH-1118 was mediated via nuclear factor kappa B (NF-κB), but not activator protein-1 (AP-1) transcription factor. This was supported by the finding that YH-1118 attenuated the phosphorylation of inhibitor of κBα (IκBα) and nuclear translocation and DNA binding activity of NF-κB. Through the mechanisms that YH-1118 inhibited the activation of IκB kinases (IKKs), upstream activators of NF-κB, or p38 MAPK, YH-1118 significantly suppressed LPS-induced production of pro-inflammatory cytokines, tumor necrosis factor-α, interleukin-1β (IL-1β), and IL-6 (p < 0.05). In conclusion, our results suggest that YH-1118 inhibits LPS-induced inflammatory responses by blocking IKK and NF-κB activation in macrophages, and may be a therapeutic candidate for the treatment of various inflammatory diseases.

• 대한한의학회지
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• 제37권2호
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• pp.62-75
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• 2016

Objectives: This study examined whether Lycii Radicis Cortex has an inhibitory effect on inflammatory response through an oxidative stress and advanced glycation endproducts (AGEs)-mediated pathway in streptozotocin (STZ)-induced type 1 diabetic rats. Methods: Lycii Radicis Cortex was orally administered to STZ-induced diabetic rats in doses of 80 or 160 mg/kg body weight/day for 2 weeks, and its effects were compared with those of diabetic control and normal rats. Results: The administration of Lycii Radicis Cortex decreased the elevated serum urea nitrogen and renal reactive oxygen species (ROS), and reduced the increased AGEs in the serum and kidney. The elevated protein expressions of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits in the kidney of diabetic control rats were significantly decreased after Lycii Radicis Cortex treatments. Moreover, the kidney of diabetic rats exhibited the up-regulation of receptor for AGEs (RAGE) and AGEs-related proteins; however, Lycii Radicis Cortex treatment also significantly reduced those expressions (excepted RAGE). In addition, the diabetic rats exhibited an up-regulation of the expression of proteins related to inflammation in the kidney, but Lycii Radicis Cortex administration reduced significantly the expression of the inflammatory proteins through the nuclear factor-kappa B (NF-${\kappa}B$) and activator protein-1 (AP-1) pathways. Conclusions: This study provides scientific evidence that Lycii Radicis Cortex exerts the antidiabetic effect by inhibiting the expressions of AGEs and NF-${\kappa}B$ in the STZ-induced diabetic rats.

• Journal of Ginseng Research
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• 제32권1호
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• pp.39-47
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• 2008

본 실험은 홍삼 혼합물 (KTNG0345)을 이용한 주름 예방 및 개선효과가 있는 건강기능 식품을 개발하기 위한 기초자료로 활용하기 위하여 시료를 경구투여한 무모생쥐의 피부조직 으로부터 MMP-3의 발현양상과 작용 메커니즘을 연구하였다. MMP-3의 발현정도는 농도 의존적으로 현저한 감소를 나타내었으며, 유전자와 단백질 모두에서 동일한 양상을 보였다. PAK는 변화가 없었지만, p38, p-p38 그리고 c-Jun, p-c-Jun 을 통계적으로 유의하게 감소시킴으로써 MMPs의 발현 감소를 가져온 것으로 보인다. 뿐만 아니라 자외선에 의한 $TNF-{\alpha}$의 생성 또는 유입을 억제함으로써 $TNF-{\alpha}$ receptor에 의해 매개되는 신호전달 경로를 둔화시켜 MMPs의 발현을 감소시킨 것으로 보인다. 이렇게 KTNG0345는 복합적인 활성으로 작용하여 주름생성 억제 활성을 보이는 것으로 판단된다.

• 생명과학회지
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• 제26권1호
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• pp.34-41
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• 2016

학명이 Aronia melanocarpa인 블랙초크베리는 항산화, 항염증, 항암 효능이 뛰어난 것으로 보고되고 있다. 본 연구에서는 블랙초크베리에 대한 collagenase inhibition effects와 산화적 스트레스에 유도된 matrix metalloproteinase(MMP), MAPkinase 그리고 AP-1의 발현 그리고/또는 인산화와 같은 분자생물학적 메카니즘을 조사하였다. Collagenase inhibition 효과는 블랙초크베리 에틸아세테이트 분획물(AE)이 500 μg/ml의 농도에서 77.2% 이상의 저해효능을 나타내었고 이는 대조군인 Epigallocatechin gallate의 결과(500 μg/ml에서 83.9%)와 비교해서 유의할 만한 결과였다. Reactive oxygen species (ROS) assay는 AE에서 가장 농도의존적으로 ROS 생성이 감소되었고, 75 μg/ml의 농도에서 약 70%로 가장 낮은 활성산소가 생성되었다. MTT assay 결과, H₂O₂에 유도된 CCRF 세포에 AE를 처치하였을 때 농도의존적으로 세포 생존율이 증가하였다. 그리고 특히, AE는 H₂O₂에 유도된 CCRF 세포에서의 MMPs (MMP-1, -3 그리고 -9), MAPK (ERK, JNK 그리고 p38) 그리고 AP-1 (c-Fos와 c-Jun)의 발현과 인산화를 억제하였고, pro-collagen type I의 발현은 증가시켰다. 따라서 블랙초크베리 에틸아세테이트 분획물은 주름억제 및 콜라겐 생성의 효능이 있으며 기능성 식품 및 화장품 소재 개발 산업에서의 응용이 가능할 것으로 기대된다.

• BMB Reports
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• 제53권6호
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• pp.323-328
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• 2020

Matrix metalloproteinase 1 (MMP-1), a calcium-dependent zinccontaining collagenase, is involved in the initial degradation of native fibrillar collagen. Tissue necrosis factor-alpha (TNFα) is a pro-inflammatory cytokine that is rapidly produced by dermal fibroblasts, monocytes/macrophages, and keratinocytes and regulates inflammation and damaged-tissue remodeling. MMP-1 is induced by TNFα and plays a critical role in tissue remodeling and skin aging processes. However, the regulation of the MMP1 gene by TNFα is not fully understood. We aimed to find additional cis-acting elements involved in the regulation of TNFα-induced MMP1 gene transcription in addition to the nuclear factor-kappa B (NF-κB) and activator protein 1 (AP1) sites. Assessments of the 5'-regulatory region of the MMP1 gene, using a series of deletion constructs, revealed the requirement of the early growth response protein 1 (EGR-1)-binding sequence (EBS) in the proximal region for proper transcription by TNFα. Ectopic expression of EGR-1, a zinc-finger transcription factor that binds to G-C rich sequences, stimulated MMP1 promoter activity. The silencing of EGR-1 by RNA interference reduced TNFα-induced MMP-1 expression. EGR-1 directly binds to the proximal region and transactivates the MMP1 gene promoter. Mutation of the EBS within the MMP1 promoter abolished EGR-1-mediated MMP-1 promoter activation. These data suggest that EGR-1 is required for TNFα-induced MMP1 transcriptional activation. In addition, we found that all three MAPKs, ERK1/2, JNK, and p38 kinase, mediate TNFα-induced MMP-1 expression via EGR-1 upregulation. These results suggest that EGR-1 may represent a good target for the development of pharmaceutical agents to reduce inflammation-induced MMP-1 expression.

• Journal of Acupuncture Research
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• 제33권3호
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• pp.29-43
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• 2016

Objectives : The purpose of this study was to investigate the effect of LR3 and SP6 acupuncture on liver damage of streptozotocin-induced diabetic mice. Methods : Male ICR mice were divided into four groups, consisting of the normal mice group(N), acupuncture-free diabetic mice group(Con), LR3-acupuncture diabetic mice group(LR3) and SP6-acupuncture diabetic mice group(SP6). The following measurements were taken: Body weight, food intake and water intake for 2 weeks; liver weight, and glucose levels in the serum and liver; ALT and AST in the serum; reactive oxygen species(ROS), reduced glutathione(GSH) and oxidized glutathione(GSSG) in the liver; and lastly, receptor for advanced glycation endproducts( RAGE), $N{\varepsilon}-carboxymethyl$ lysine(CML), $N{\varepsilon}-carboxyethyl$ lysine(CEL), phosphorylation of inhibitory kappa B alpha($p-I{\kappa}B{\alpha}$), nuclear factor-kappa B($NF-{\kappa}B$), activator protein-1(AP-1), cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), tumor necrosis factor-alpha($TNF-{\alpha}$), ${\beta}-actin$, cytochrome c and caspase in the liver. Results : The liver weight and GSH/GSSG ratio were significantly increased in SP6 compared to Con. The glucose levels in the liver were significantly decreased in LR3 compared to Con. The generation of ROS and GSSG were significantly decreased in SP6 compared to Con. The expressions of RAGE, CML, AP-1, $TNF-{\alpha}$, cytochrome c and caspase 3 were significantly decreased in LR3 compared to Con. The expressions of $p-I{\kappa}B{\alpha}$, $NF-{\kappa}B$, AP-1, COX-2, iNOS and caspase 3 were significantly decreased in SP6 compared to Con. Conclusion : It is predicted that LR3 acupuncture is related to reduced glucose levels in the liver and expressions of AGE, and that, SP6 acupuncture is related to reduced oxidative stress-related transcription factors and inflammation-related proteins. Therefore, we suggest that LR3 and SP6 acupuncture have protective effects on the liver of streptozotocin-induced diabetic mice by preventing apoptosis.

• 대한의생명과학회지
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• 제14권3호
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• pp.147-155
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• 2008

In previous works, we found that solvent extract of Opuntia humifusa Raf., a member of the lactaceae family, displayed potent anti-oxidative and anti-inflammatory activities. Thus, all solvent fractions, except for the water layer, showed potent scavenging effects. According to activity-guided fractionation, one of active radical scavenging principles in the ethyl acetate fraction was found to be taxifolin. In this study, we investigated whether taxifolin showed anti-oxidative activity. In addition, taxifolin modulated nitric oxide (NO) release and the expression of pro-inflammatory cytokine mRNA such as interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-${\alpha}$. Taxifolin showed potent anti-oxidant activity with the $IC_{50}\;of\;8.5{\pm}1.4\;and\;9.3{\pm}1.0{\mu}M$ using xanthine/xanthine oxidase (XO) assay and 2,2-Diphenyl-lpicrylhydrazyl radical (DPPH) assay, respectively. We next determined the role of taxifolin on the immunomodulating activity using murine macrophage cell line RAW264.7 cells. Taxifolin dose-dependently inhibited NO production in lipopolysaccharide (LPS)-activated RAW264.7. It also significantly blocked the expression of inducible NO synthase (iNOS) mRNA in the LPS-stimulated RAW264.7 cells. In addition, taxifolin potently suppressed the expression of IL-$1{\beta}$, IL-6 and GM-CSF mRNA in LPS-activated RAW264.7 cells, but not that of TNF-${\alpha}$ Moreover, taxifolin significantly inhibited the transcriptional activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and activator protein -1 (AP-1). These results suggest that taxifolin may downregulate inflammatory iNOS, IL-$1{\beta}$, IL-6 and GM-CSF gene expressions through inhibition of NF-K and AP-1 activation in LPS-stimulated RAW264.7 cells.

• 대한한방내과학회지
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• 제21권2호
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• pp.259-266
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• 2000

Objectives : The effects of aqueous extracts of Cortex ulmi pumilae (a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origm hepatoma cell lines, HepG2. Methods : The death of HepG2 cells was markedly induced by the addition of extracts of Cortex ulmi pumilae in a dose-dependent manner. The apoptotic characteristic ladder pattern of DNA strand break was not observed in cell death of HepG2. In addition, it was not shown nucleus chromatin condensation and fragmentation under hoechst staining. However, by the using annexin V staining assay, externalizations of phosphatidylserine in HepG2 cell which were treated with Cortex ulmi pumilae extracts were detected in the early time (at 9 hr after extract treatment). Furthermore, LDH release was not detected in this early stage. Therefore, Cortex ulmi pumilae extracts-induced cell death of HepG2 cells is mediated by apoptotic death signal processes. Result : The activity of caspase 3-like proteases remained in a basal level in HepG2 cells which treated with the extract of Cordyceps sinensis. However, it was markedly increased in HepG2 cells which treated with two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) which were differently extracted (respectively, 2.3 and 3.3 fold). On a while, the phosphotransferase activities of JNK1 was markedly induced in HepG2 cells which were treated with two extracts of Cortex ulmi pumilae. On the contrary, the activation of transcriptional activator, activating protein1(AP-1) and NF-kB were severely decreased by these two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K). In addition, antioxidants (GSH and NAC) and intracellular $Ca2^+$ level regulator (Bapta/AM and Thapsigargin) did not affect Cortex ulmi pumilae extracts-induced apoptotic death of HepG2 cells. Conclusions : In conclusion, our results suggest that two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) induces the apoptotic death of human liver origin hepatoma HepG2 cells via activation of caspase 3-like proteases as well as JNK1, and inhibition of transcriptional activators, AP-1 and $NK-{\kappa}B$.

• The Korean Journal of Physiology and Pharmacology
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• 제14권1호
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• pp.29-35
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• 2010

We have shown that myosin light chain kinase (MLCK) was required for the off-contraction in response to the electrical field stimulation (EFS) of feline esophageal smooth muscle. In this study, we investigated whether protein kinase C (PKC) may require the on-contraction in response to EFS using feline esophageal smooth muscle. The contractions were recorded using an isometric force transducer. On-contraction occurred in the presence of $N^G$-nitro-L-arginine methyl ester (L-NAME), suggesting that nitric oxide acts as an inhibitory mediator in smooth muscle. The excitatory composition of both contractions was cholinergic dependent which was blocked by tetrodotoxin or atropine. The on-contraction was abolished in $Ca^{2+}$-free buffer but reappeared in normal $Ca^{2+}$-containing buffer indicating that the contraction was $Ca^{2+}$ dependent. 4-aminopyridine (4-AP), voltage-dependent $K^+$ channel blocker, significantly enhanced on-contraction. Aluminum fluoride (a G-protein activator) increased on-contraction. Pertussis toxin (a $G_i$ inactivator) and C3 exoenzyme (a rhoA inactivator) significantly decreased on-contraction suggesting that Gi or rhoA protein may be related with $Ca^{2+}$ and $K^+$ channel. ML-9, a MLCK inhibitor, significantly inhibited on-contraction, and chelerythrine (PKC inhibitor) affected on the contraction. These results suggest that endogenous cholinergic contractions activated directly by low-frequency EFS may be mediated by $Ca^{2+}$, and G proteins, such as Gi and rhoA, which resulted in the activation of MLCK, and PKC to produce the contraction in feline distal esophageal smooth muscle.