• Bulletin of the Korean Chemical Society
• /
• 제29권9호
• /
• pp.1717-1722
• /
• 2008

The 70 kDa heat-shock protein (Hsp70) involved in various cellular functions, such as protein folding, translocation and degradation, regulates apoptosis in cancer cells. Recently, it has been reported that the green tea flavonoid (−)-epigallocatechin 3-gallate (EGCG) induces apoptosis in numerous cancer cell lines and could inhibit the anti-apoptotic effect of human Hsp70 ATPase domain (hATPase). In the present study, docking model between EGCG and hATPase was determined using automated docking study. Epi-gallo moiety in EGCG participated in hydrogen bonds with side chain of K71 and T204, and has metal chelating interaction with hATPase. Hydroxyl group of catechin moiety also participated in metal chelating hydrogen bond. Gallate moiety had two hydrogen bondings with side chains of E268 and K271, and hydrophobic interaction with Y15. Based on this docking model, we determined two pharmacophore maps consisted of six or seven features, including three or four hydrogen bonding acceptors, two hydrogen bonding donors, and one lipophilic. We searched a flavonoid database including 23 naturally occurring flavonoids and 10 polyphenolic flavonoids with two maps, and myricetin and GC were hit by map I. Three hydroxyl groups of B-ring in myricetin and gallo moiety of GC formed important hydrogen bonds with hATPase. 7-OH of A-ring in myricetin and OH group of catechin moiety in GC are hydrogen bond donors similar to gallate moiety in EGCG. From these results, it can be proposed that myricetin and GC can be potent inhibitors of hATPase. This study will be helpful to understand the mechanism of inhibition of hATPase by EGCG and give insights to develop potent inhibitors of hATPase.

• BMB Reports
• /
• 제37권2호
• /
• pp.239-245
• /
• 2004

We have previously demonstrated that the redox reactant pyruvate prevents apoptosis in the oxidant model of bovine pulmonary artery endothelial cells (BPAEC), and that the anti-apoptotic mechanism of pyruvate is mediated in part via the mitochondrial matrix compartment. However, cytosolic mechanisms for the cytoprotective feature of pyruvate remain to be elucidated. This study investigated the pyruvate protection against endothelial cytotoxicity when the glycolysis inhibitor 2-deoxy-D-glucose (2DG) was applied to BPAEC. Millimolar 2DG blocked the cellular glucose uptake in a concentration- and time-dependent manner with >85% inhibition at $\geq$5 mM within 24 h. The addition of 2DG evoked BPAEC cytotoxicity with a substantial increase in lipid peroxidation and a marked decrease in intracellular total glutathione. Exogenous pyruvate partially prevented the 2DG-induced cell damage with increasing viability of BPAEC by 25-30%, and the total glutathione was also modestly increased. In contrast, 10 mM L-lactate, as a cytosolic reductant, had no effect on the cytotoxicity and lipid peroxidation that are evoked by 2DG. These results suggest that 2DG toxicity may be a consequence of the diminished potential of glutathione antioxidant, which was partially restored by exogenous pyruvate but not L-lactate. Therefore, pyruvate qualifies as a cytoprotective agent for strategies that attenuate the metabolic dysfunction of the endothelium, and cellular glucose oxidation is required for the functioning of the cytosolic glutathione/NADPH redox system.

• Biomolecules & Therapeutics
• /
• 제30권4호
• /
• pp.380-388
• /
• 2022

Snail is implicated in tumour growth and metastasis and is up-regulated in various human tumours. Although the role of Snails in epithelial-mesenchymal transition, which is particularly important in cancer metastasis, is well known, how they regulate tumour growth is poorly described. In this study, the possible molecular mechanisms of Snail in tumour growth were explored. Baculoviral inhibitor of apoptosis protein (IAP) repeat-containing protein 3 (BIRC3), a co-activator of cell proliferation during tumourigenesis, was identified as a Snail-binding protein via a yeast two-hybrid system. Since BIRC3 is important for cell survival, the effect of BIRC3 binding partner Snail on cell survival was investigated in ovarian cancer cell lines. Results revealed that Bax expression was activated, while the expression levels of anti-apoptotic proteins were markedly decreased by small interfering RNA (siRNA) specific for Snail (siSnail). siSnail, the binding partner of siBIRC3, activated the tumour suppressor function of p53 by promoting p53 protein stability. Conversely, BIRC3 could interact with Snail, for this reason, the possibility of BIRC3 involvement in EMT was investigated. BIRC3 overexpression resulted in a decreased expression of the epithelial marker and an increased expression of the mesenchymal markers. siSnail or siBIRC3 reduced the mRNA levels of matrix metalloproteinase (MMP)-2 and MMP-9. These results provide evidence that Snail promotes cell proliferation by interacting with BIRC3 and that BIRC3 might be involved in EMT via binding to Snail in ovarian cancer cells. Therefore, our results suggested the novel relevance of BIRC3, the binding partner of Snail, in ovarian cancer development.

• 동의생리병리학회지
• /
• 제36권6호
• /
• pp.229-234
• /
• 2022

Lung cancer is the leading cause of cancer-related deaths worldwide. Indigo Naturalis (IN) is a dark blue powder obtained by processing leaves or stems of indigo plants, its anticancer effects have been reported in several studies. However, the pharmacological mechanism of IN in small cell lung cancer (SCLC) is not elucidated. In this study, to investigate the anticancer efficacy of IN for SCLC, we presented potential active ingredients, SCLC-related targets, and pharmacological mechanisms of IN that are expected to have anticancer activity for SCLC using a network pharmacological analysis. The phytochemical compounds of IN have been collected through TCMSP, SymMap, or HPLC documents. The active ingredients of IN such as indirubin, indican, isatin, and tryptanthrin were selected through ADME parameters or literature investigations for each compound. Using the Compounds, Disease-Target associations Databases, 124 common targets of IN and SCLC were obtained. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway enrichment analysis was carried out. GO biological processes are associated with response to xenobiotic stimulus, positive regulation of protein phosphorylation, regulation of mitotic cell cycle, and regulation of apoptotic signaling pathway. KEGG disease pathways included Gastric cancer, Bladder cancer, SCLC, and Melanoma. The main anticancer targets of the IN for SCLC were analyzed in 14 targets, including BCL2, MYC, and TP53. In conclusion, the results of this study based on the network pharmacology of IN can provide important data for the effective prevention and treatment of SCLC.

• 한국임상수의학회지
• /
• 제41권1호
• /
• pp.49-53
• /
• 2024

A two-year-old, spayed female, 22.5 kg Pungsan was referred with chronic crusts and erosion on the nose. A referring veterinarian prescribed an anti-inflammatory dose (0.5-1 mg/kg/day) of oral glucocorticoids for 5 months, but skin lesions showed no meaningful improvement. A dermatological evaluation revealed a crust, depigmentation, erosion, and erythematous lesion over the nasal planum with a loss of the normal cobblestone texture. Also, firm and multifocal plaques over the thigh, groin, axilla, and dorsum were detected. A cytology examination on the nose, thigh, groin, axilla, and dorsum revealed moderate neutrophilic inflammation and bacterial infection. Abdominal radiography and ultrasonography revealed subcutaneous calcified materials along the thigh, groin, axilla, and dorsum. Calcinosis cutis was suspected because of the adverse effect of previous prolonged corticosteroid therapy. A histopathology examination of the nose lesion revealed moderate to severe degenerative or apoptotic changes of the basal layer and lymphoplasmacytic interface dermatitis. Facial discoid lupus erythematosus (FDLE) was diagnosed based on the history and the clinical, cytological, and histopathological results. Minocycline (7 mg/kg PO q 12 h) and niacinamide (500 mg/dog PO q 12 h) were prescribed as initial treatment. Glucocorticoids were not administered due to the presence of calcinosis cutis induced by previous corticosteroid treatment. After 6 weeks of treatment, the clinical signs on the nose were mildly improved. At this time, topical 1% pimecrolimus cream (twice daily) was initiated, while minocycline and niacinamide were continued at the same dose. The nasal planum markedly improved after 6 weeks of additional treatment, hence minocycline and niacinamide were prescribed for an additional 2 weeks and stopped, and the patient was continued solely on topical pimecrolimus. The dog's skin lesion has been maintained in clinical remission with topical 1% pimecrolimus twice daily for more than 5 months.

• IMMUNE NETWORK
• /
• 제23권2호
• /
• pp.14.1-14.14
• /
• 2023

Immune status including the immune cells and cytokine profiles has been implicated in the development of endometriosis. In this study, we analyzed Th17 cells and IL-17A in peritoneal fluid (PF) and endometrial tissues of patients with (n=10) and without (n=26) endometriosis. Our study has shown increased Th17 cell population and IL-17A level in PF with endometriosis patients. To determine the roles of IL-17A and Th17 cells in the development of endometriosis, the effect of IL-17A, major cytokine of Th17, on endometrial cells isolated from endometriotic tissues was examined. Recombinant IL-17A promoted survival of endometrial cells accompanied by increased expression of anti-apoptotic genes, including Bcl-2 and MCL1, and the activation of ERK1/2 signaling. In addition, treatment of IL-17A to endometrial cells inhibited NK cell mediated cytotoxicity and induced HLA-G expression on endometrial cells. IL-17A also promoted migration of endometrial cells. Our data suggest that Th17 cells and IL-17A play critical roles in the development of endometriosis by promoting endometrial cell survival and conferring a resistance to NK cell cytotoxicity through the activation of ERK1/2 signaling. Targeting IL-17A has potential as a new strategy for the treatment of endometriosis.

• 한국축산식품학회지
• /
• 제44권3호
• /
• pp.699-709
• /
• 2024

Oxya chinensis sinuosa (OC) is a well-known edible insect. Several researches on the health benefits of OC consumption have been performed to date; however, their effect on eye health remains largely unknown. This study aimed to assess the protective effects of OC extracts on the oxidative stress on the retinal pigment epithelium (RPE) cells. Oxidative damage has been identified as one of the key regulatory factors in agerelated macular degeneration. H₂O₂-induced reactive oxygen species (ROS) production, a well-known oxidative stress factor, can cause cell death in retinal pigment epithelia cells. In this study, we found that three OC extracts effectively prevented H₂O₂-induced ROS production and subsequent death of ARPE-19 cells in a dose-dependent manner. In addition, the OC extracts inhibited the phosphorylation of mitogen-activated protein kinases including p38, JNK, and ERK. The OC extracts restored IκBα degradation induced by H₂O₂, indicating that OC extracts suppressed the activation of nuclear factorκB. Furthermore, the three OC extracts were shown to have antioxidant effects by upregulating the intracellular expression of key antioxidant proteins such as SOD, NQO, and HO-1. Here we demonstrated the antioxidant and anti-apoptotic effects of the OC extracts on ARPE-19, indicating their potential role in improving eye health. These results suggest that three OC extracts plays a critical role in oxidative stress-induced cell death protects in ARPE-19 cells.

• Journal of Nutrition and Health
• /
• 제38권10호
• /
• pp.807-816
• /
• 2005

본 연구에서는 Sprague Dawley종 수컷 쥐에서 DHM로 대장암을 유발시킨 후 식이에 첨가한 n-3 RUFA 함량에 따라 대장의 암화과정에 어떤영향을 미치는지 생화학적인 기전을 검토하여 다음과 같은 결과를 얻었다. 1) 식이에 n-3 RUFA를 6.2 mmole 첨가하여 먹인 경우에는 COX-2의 mRNA와 단백질의 발현 및 TBX2와 PGE2 합성이 유의하게 감소 되었으며, 이에 따라 세포증식도 억제되었다. 그러나 n-3 RUFA를 12.4 mmole 첨가한 식이로 사육한 경우에는 오히려 COX-2의 mRNA와 단백질의 발현 및 $TXB_2$ 와 $PGE_2$ 합성이 대조군 보다도 더 많이 증가되었고 세포증식도 대조군과 같은 수준으로 증가하여 암화과정을 촉진하였다. 2) 식이에 n-3 PUFA를 6.2 mmole 첨가한 경우 대장 점막의 인지질의 지방산조성중 arachidonic 함량은 유의하게 낮았고, EPA와 DHA 함량은 유의하게 높았으며 n-3 PUFA를 12.4 mmole 첨가한 경우에는 EPA와 DHA 함량은 2배 이상 유의하게 더 높았다. 3) 대장 상피세포의 apoptosis는 식이에 n-3 PUFA를 첨가한 함량과는 관계없이 대조군에 비해 유의하게 약 34$\sim$$42\%$ 증가하였다. 4) 식이에 n-3 PUFA를 6.2 mmole 첨가하였을 때 Bax의 mRNA와 단백질 발현은 각각 유의하게 증가되었으나 n-3 PUFA를 12.4mmole 첨가한 경우에는 유의하게 감소되었다. 반면에Bcl-2의 mRNA와 단백질 발현은 n-3 PUFA 6.2 mmole 첨가한 경우에는 오히려 증가하였다. 총괄해서 본 연구결과에 의하면 항암효과가 있가도 알려진 n-3 PUFA는 식이에 첨가한 함량에 따라 암화과정에 미치는 효과가 다르게 나타났다, 그러므로 앞으로 대장의 암화과정을 억제시키는 n-3 PUFA의 적절량 또는 섭취의 한계치를 정하는 연구가 필요하다고 본다.

• 동의생리병리학회지
• /
• 제23권4호
• /
• pp.818-824
• /
• 2009

This study aimed to evaluate the effect of purgation therapy with Natrii sulfas, a therapy for stroke patients with constipation in the oriental medicine, on the ischemic brain damage of the rats. The ischemic brain damage was induced by the middle cerebral artery occlusion (MCAO), Natrii sulafas was administered once after the MCAO. After 48 hours, expressions of Bax, Bcl-2, c-Fos, and HSP72 on the brain tissues were observed by immunohistochemistrical methods or technique. Purgation therapy with Natrii sulfas attenuated the excess of Bax expression caused by the ischemic brain damage. It was significant statistically in the penumbra of cerebral cortex, but not in the caudate putamen, of the MCAO rats. Purgation therapy with Natrii sulfas did not attenuate the excess of Bcl-2 expression caused by the ischemic brain damage. Purgation therapy with Natrii sulfas did not attenuate the excess of c-Fos expression caused by the ischemic brain damage. Purgation therapy with Natrii sulfas attenuated the excess of HSP72 expression caused by the ischemic brain damage. It was significant statistically in the penumbra of cerebral cortex, but not in the caudate putamen, of the MCAO rats. These results suggest that purgation therapy with Natrii sulfas has a neuroprotective effect on the ischemic brain damage and an anti-apoptotic effect.

• 대한치과교정학회지
• /
• 제33권6호
• /
• pp.453-463
• /
• 2003

본 연구는 MC3T3El 조골세포가 저산소증에 반응하여 유발될 수 있는 세포 고사조절 기전을 구명하고자 함에 목적이 있다. $2\%$ 저산소증의 조건하에서 MC3T3El 조골세포는 DNA 사다리 분절 헝성을 보였으며 형광성 염료인 Hoechst 33258로 염색된 핵 구조 형태 관찰시 시간이 지남에 따라 세포고사 현상을 관찰할 수 있었다 Pancaspase 억제제인 Z-VAD-FMK나 특정한 caspase-3 억제제인 Z-DEVD-CHO로 사전 처치하였을 경우에는 저산소증에 의한 DNA 사다리 분절형성이 농축에 비례하여 억제되었다. caspase-3류의 프로테아제(DEVDase) 활성 증가가 세포고사 중에 관찰되었으나 caspase-1 (YVADase)의 활성은 없었다. 어떤 caspase가 세포고사에 관여하는지를 확인하기 위하여 anti-caspase-3 또는 anti-caspase-6의 항체를 이용한 western blotting이 시행되었다. caspase-3의 활성산물에 해당하는 17-KDa단백질과 caspase-6의 활성산물인 20-KDa 단백질이 세포용해물에서 발생되었다. 또한 시간 경과와 더불어 caspase-6의 활동의 상징인 Lamin A의 분열을 일으켰으며, 사이토크롬 C를 cytosol로 방출하였다. 이로써 저산소증에 의한 조골세포의 고사 과정에 사이토크롬 C의 방출이 포함된 caspase의 활성이 관여한다는 것을 확인할 수 있었다.