• Biomolecules & Therapeutics
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• 제32권2호
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• pp.240-248
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• 2024

We observed that treatment with dimethyl α-ketoglutarate (DMK) increased the amount of intracellular α-ketoglutarate significantly more than that of α-ketoglutarate in HaCaT cells. DMK also increased the level of intracellular 4-hydroxyproline and promoted the production of collagen in HaCaT cells. In addition, DMK decreased the production of collagenase and elastase and down-regulated the expression of selected matrix metalloproteinases (MMPs), such as MMP-1, MMP-9, MMP-10, and MMP-12, via transcriptional inhibition. The inhibition of MMPs by DMK was mediated by the suppression of the IL-1 signaling cascade, leading to the attenuation of ERK1/2 phosphorylation and AP-1 transactivation. Our study results illustrate that DMK, an alkylated derivative of α-ketoglutarate, increased the level of 4-hydroxyproline, promoted the production of collagen, and inhibited the expression of selected MMPs by affecting the IL-1 cascade and AP-1 transactivation in HaCaT cells. The results suggest that DMK might be useful as an anti-wrinkle ingredient.

• Journal of Microbiology and Biotechnology
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• 제34권7호
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• pp.1401-1409
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• 2024

Postbiotics have various functional effects, such as antioxidant, anti-inflammatory, and anti-obesity. Levilactobacillus brevis BK3, the subject of this study, was derived from lactic acid bacteria isolated from Kimchi, a traditional Korean fermented food. The antioxidant activity of BK3 was confirmed through the measurements of 2,2-diphenyl-1-picryl-hydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and total antioxidant capacity (TAC). The wrinkle improvement effect was validated by assessing elastase inhibitory activity and collagenase inhibitory activity. The intracellular activity was confirmed using human keratinocytes (HaCaT) and human fibroblasts (HFF-1). BK3 protects skin cells from oxidative stress induced by H₂O₂ and reduces intracellular reactive oxygen species (ROS) production. In addition, the expressions of the antioxidant genes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were upregulated. Meanwhile, matrix metalloproteinase-1 (MMP-1) and collagen type I alpha 1 (COL1A1), involved in collagen degradation and synthesis, were significantly regulated. These results suggest the possibility of utilizing BK3 as a functional ingredient with antioxidant and wrinkle-improving effects.

• Parasites, Hosts and Diseases
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• 제54권2호
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• pp.139-145
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• 2016

Toxoplasmosis is a serious disease caused by Toxoplasma gondii, one of the most widespread parasites in the world. Lipid metabolism is important in the intracellular stage of T. gondii. Stearoyl-CoA desaturase (SCD), a key enzyme for the synthesis of unsaturated fatty acid is predicted to exist in T. gondii. Sterculic acid has been shown to specifically inhibit SCD activity. Here, we examined whether sterculic acid and its methyl ester analogues exhibit anti-T. gondii effects in vitro. T. gondii-infected Vero cells were disintegrated at 36 hr because of the propagation and egress of intracellular tachyzoites. All test compounds inhibited tachyzoite propagation and egress, reducing the number of ruptured Vero cells by the parasites. Sterculic acid and the methyl esters also inhibited replication of intracellular tachyzoites in HFF cells. Among the test compounds, sterculic acid showed the most potent activity against T. gondii, with an $EC_{50}$ value of $36.2{\mu}M$, compared with $EC_{50}$ values of $248-428{\mu}M$ for the methyl esters. Our study demonstrated that sterculic acid and its analogues are effective in inhibition of T. gondii growth in vitro, suggesting that these compounds or analogues targeting SCD could be effective agents for the treatment of toxoplasmosis.

• Restorative Dentistry and Endodontics
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• 제24권3호
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• pp.437-446
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• 1999

Porphyromonas endodontalis(P. endodontalis) is one of the important causative bacteria of pulpal and periapical disease. P. endodontalis has lipopolysaccharide(LPS) and it plays a major role in stimulating the synthesis and release of cytokines from immune cells and prostaglandin $E_2$ from host cells. The purpose of this study is to prepare LPS from P. endodontalis and to evaluate the effect of LPS on membrane permeability of fibroblast. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted. LPS was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Human periodontal ligament cell, colon fibroblast(CCD-18Co, KCLB 21459) and skin fibroblast(Detroit 551, KCLB 10110) were perfused with 0.01% P. endodontalis LPS solution, high concentration of $K^+$ solution and $Ca^{2+}$-free solution, $Ca^{2+}$ concentration ratio was measured by microfluorometry. 1. Intracellular $Ca^{2+}$ concentration was not changed in human periodontal fibroblast and skin fibroblast(Detroit 551) stimulated by P. endodontalis LPS. 2. Intracellular $Ca^{2+}$ concentration was increased in colon fibroblast(CCD-18Co) stimulated by P. endodontalis LPS. 3. Colon fibroblast(CCD-18Co) has voltage dependent $Ca^{2+}$ channel activated by high concentration of $K^+$ solution. 4. P. endodontalis LPS has no effect on the increase of intracellular $Ca^{2+}$ concentration during perfusion of $Ca^{2+}$-free solution.

• The Korean Journal of Physiology and Pharmacology
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• 제25권1호
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• pp.87-94
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• 2021

Skin photoaging occurs due to chronic exposure to solar ultraviolet radiation (UV), the main factor contributing to extrinsic skin aging. Clinical signs of photoaging include the formation of deep, coarse skin wrinkles and hyperpigmentation. Although melanogenesis and skin wrinkling occur in different skin cells and have different underlying mechanisms, their initiation involves intracellular calcium signaling via calcium ion channels. The ORAI1 channel initiates melanogenesis in melanocytes, and the TRPV1 channel initiates MMP-1 production in keratinocytes in response to UV stimulation. We aimed to develop a drug that may simultaneously inhibit ORAI1 and TRPV1 activity to help prevent photoaging. We synthesized nootkatol, a chemical derivative of valencene. TRPV1 and ORAI1 activities were measured using the whole-cell patch-clamp technique. Intracellular calcium concentration [Ca2+]i was measured using calcium-sensitive fluorescent dye (Fura-2 AM). UV-induced melanin formation and MMP-1 production were quantified in B16F10 melanoma cells and HaCaT cells, respectively. Our results indicate that nootkatol (90 μM) reduced TRPV1 current by 94% ± 2% at -60 mV and ORAI1 current by 97% ± 1% at -120 mV. Intracellular calcium signaling was significantly inhibited by nootkatol in response to ORAI1 activation in human primary melanocytes (51.6% ± 0.98% at 100 μM). Additionally, UV-induced melanin synthesis was reduced by 76.38% ± 5.90% in B16F10 melanoma cells, and UV-induced MMP-1 production was reduced by 59.33% ± 1.49% in HaCaT cells. In conclusion, nootkatol inhibits both TRPV1 and ORAI1 to prevent photoaging, and targeting ion channels may be a promising strategy for preventing photoaging.

• 한국식품영양과학회지
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• 제43권5호
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• pp.705-711
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• 2014

피부미백 소재를 개발하기 위해 멜라닌 함량, 세포내 tyrosinase 활성의 측정 및 Western blotting 실험이 수행되었다. 백합(Lilium Oriental Hybrid 'Siberia') 뿌리의 80% 에탄올 추출물로부터 얻은 에틸아세테이트 분획물(R-EA)은 ${\alpha}$-melanocyte stimulating hormone(${\alpha}$-MSH)에 의해 멜라닌 생성이 유도된 B16/F10 흑색종 세포에서 농도 의존적으로 멜라닌 생성을 저해하였다. 정확하게 세포내 tyrosinase 활성과 멜라닌 함량은 에틸아세데이트 분획물 $100{\mu}g/mL$ 처리 시 ${\alpha}$-MSH 단독 처리군에 비해 각각 45%와 74%의 저해율을 보였다. ${\alpha}$-MSH에 의해 멜라닌 생성이 유도된 B16/F10 흑색종 세포에서 단백질 발현양상을 살펴본 결과 TRP-1이 가장 많이 억제된 양상을 확인할 수 있었고 이 결과는 세포내 tyrosinase 활성저해보다 멜라닌 생성저해가 더 많이 일어난 것과 일맥상통하는 것이다. 이를 종합해 볼 때 p-coumaric acid와 resveratrol 함량이 백합뿌리의 에탄올 추출물에 비해 많이 함유된 에틸아세테이트 분획물은 멜라닌 생성 유도물질에 의해 촉진된 ERK의 활성화를 억제하는 피부미백 소재로서 그 가치가 입증된다고 사료된다.

• 한국미생물·생명공학회지
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• 제19권2호
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• pp.128-134
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• 1991

To study the reduced growth and synthesis, proeviously reported, of ${\beta}$-galactosidase of alkalophilic Bacillus sp. YS-309 at the higher lactose concentration of 0.5% (w/v) in the medium, lactose transport and utilization were examined. The results showed that lactose transport was influenced by the addition of four kinds of antibiotics, and tetracycline stimulated most but not valinomycin. PEP-potentials of the cells grown on lactose was estimated lower than the cells on glucose and on galactose. Thus, the transport of lactose was independent of intracellular PEP and phosphorylation reactions, and was thought to be uptaked directly or oxidized in part in the transport process. In the other hand, once lactose was uptaked into the cells, it was hydrolyzed by ${\beta}$-glactosidase to glucose and galactose. The former was metabolized fast but the latter was accumulated. Galactose and lactose were not utilized until glucose was mostly depleted in the medium. The ${\beta}$-galactosidase synthesis decreased in the presence of glucose over 0.2% and galactose over 0.05 to 0.1%, respectively. In conclusion, it was considered for glucose as a repressor and galactose as a inducer for ${\beta}$-galactosidase synthesis even though the mechanisms were not elucidated. Catabolite repression of glucose on the enzyme synthesis was not relieved by the addition of exogeneous cAMP.

• 한국안광학회지
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• 제8권1호
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• pp.65-71
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• 2003

각막 상피세포는 정상적인 apoptosis과정을 거쳐 세포가 탈락하고 재생한다. 이러한 apoptosis에는 많은 요소들이 관여하여 세포가 사멸하는데 여러 가지 메커니즘이 관여한다. 본 연구에서는 세포고사 인자로 알려진 물질들을 각막 상피세포에서 apoptosis의 유도 여부를 다른 세포와 비교하여 각막 상피세포의 특성을 알아보고자 시행하였다. 본 연구에 이용된 세포고사 유도물질은 recombinant human cytokiness ($INF{\gamma}$, $TNF{\alpha}$, FASAb), actinomycin D. camptothecin, cycloheximide, dexamethasone와 etoposide이다. 이들을 세포에 48시간 처리한 후 세포독성을 MTT assay로 측정하였으며 세포고사는 Hoechst 33342 staining. Annexin V-FITC/PI staining 그리고 DePsipher assay를 이용하였다. 세포고사의 한 경로인 FAS-FAS ligand system에 대한 연구는 immunocytochemistry로 Fas protein 발현 여부를 조사하였다. 모든 유도인자는 농도의존적으로 세포고사를 유도하였는데 Actinomycin D. camptothecin와 etoposide는 제조사의 추천 농도보다 낮은 농도에서 세포고사가 유도되었고 반면에 cytokines, cycloheximide, dexamethasone은 더 높은 농도에서 세포고사를 유도하였다. FAS antigen은 대조군과 처리군 모두에서 발현되었으나 세포고사율에 비례하여 높게 발현되었다. 본 연구 결과 각막 상피세포는 RNA synthesis inhibitor와 topoisomerase inhibitors가 intracellular receptor-activators 보다 세포고사에 민감하게 나타나는 세포의 특성을 보였다.

• Korean Journal of Acupuncture
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• 제37권2호
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• pp.88-96
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• 2020

Objectives : The purpose of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Euphorbia lathyris L. extracted by supercritical CO₂. Methods : Human dermal fibroblast cells dosed with the extract from Euphorbia lathyris L. were harvested and the intracellular proteome was analyzed to examine the expression of proteins related collagen synthesis pathway, metalloproteinases (MMPs), extracellular matrix (ECM)-cell interaction, cytokines, and antioxidant enzymes by 2-dimensional gel electrophoresis. Results : Fatty acid analysis of the extract from Euphorbia lathyris L. showed oleic acid was 84% and linoleic acid was 4.1%. Antioxidative effect was about 53% by beta carotene bleaching assay. In 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis, fifteen protein changes in five mechanisms which were collagen synthesis pathway, MMPs, ECM-cell interaction, cytokines, and antioxidant enzymes were analyzed. Conclusions : This study suggests the supercritical extraction from the seed of Euphorbia lathyris L. could be used as anti-oxidant substances for pharmacopuncture.

• Journal of Microbiology
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• 제34권3호
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• pp.253-269
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• 1996

Infection of fish cells with IHNV resulted in gradual increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/)] in CHSE, gradual decrease in [Ca$\^$2+/] in FHM, and no significant change in RTG cells. The degree of [Ca$\^$2+/] increase or decrease was dependent on the amount of infectious virus, and these [Ca$\^$2+/] variations were maximal at 16 hours after virus infection (p. i.) in both cell lines. When the fish cells were infected with inactivated IHNV, evident variation in [Ca$\^$2+/] was not observed. Thus, infectivity of IHNV appears to correlate with changes in [Ca$\^$2+/] in virus-infected cells. These IHNV-induced [Ca$\^$2+/] changes were partially blocked by cycloheximide, but not affected by cordycepin. It seems to be that virus-induced Ca$\^$2+/ variations were more related with protein synthesis than RNA synthesis. Various Ca$\^$2+/ related drugs were used in search for the mechanisms of the [Ca$\^$2+/], changes following IHNV infection of CHSE cells. Decreasing extracellular Ca$\^$2+/ concentration or blocking Ca$\^$2+/ influx from extracellular media inhibited the IHNV-induced increase in [Ca$\^$2+/], in CHSE cells. Similar results were obtained with intracellular Ca$\^$2+/ sources are important in IHNV-induced [Ca$\^$2+/] increase in CHSE cells.