• Biomolecules & Therapeutics
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• 제9권3호
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• pp.201-208
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• 2001

The present study was conducted to examine the effects of cholinergic stimulation and membrane depolarization on secretion of epinephrine (EP) and norepinephrine (NE) in the perfused model of the rat adrenal gland and to investigate the effect of bromocriptine on secretion of EP and NE evoked by these secreta-gogues. Acetylcholine (ACh, 5.32 mM), high $K^{+}$(56mM), 1.1-dimethyl-4-phenyl piperazinium iodide (DMPP, 100 $\mu$M for 2 min), (3-(m-cholro-phenyl-carbamoyl-oxy)-2butynyl trimethyl ammonium chloride (McN-A-343, 100 $\mu$M for 2 min), cyclopiazonic acid (10 $\mu$M for 4 min) and methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2-trifluoromethylphenyl) -pyridine-5-carboxylate (Bay-K-8644, 10 $\mu$M for 4 min) evoked a 1.3~5.3-fold greater secretion of EP than NE in the perfused rat adrenal gland. The perfusion of bromocriptine (1-10 $\mu$M) into an adrenal vein for 20 min produced relatively dose-dependent inhibition in secretion of EP and NE evoked by ACh, high $K^{+}$, DMPP, and McN-A-343. Moreover, under the presence of bromocriptine (1~10 $\mu$M), releasing responses of EP and NE evoked by cyclopiazonic acid and Bay-K-8644 were also greatly reduced. Taken together, these results suggest that cholinergic stimulation and membrane depolarization enhance more release of EP than NE in the perfumed rat adrenal medulla, and that bromocriptine inhibits the release of EP and NE evoked by stimulation of cholinergic receptors as well as by membrane depolarization. It seems that this inhibitory effect of bromocriptine is associated with inhibition of calcium channels through activation of dopaminergic D2-receptors located in the rat adrenomedullary chromaffin cells.lls.

• Biomolecules & Therapeutics
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• 제8권1호
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• pp.13-21
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• 2000

The present study was designed to investigate the effect f quinidine on catecholamine (CA) secretion evoked by ACh, high $K^{+}$, DMPP, McN-A343, cyclopiazonic acid and Bay-K-8644 from the isolated perfused rat adrenal gland and to establish the mechanism of its action. The perfusion of quinidine (15-150 $\mu$M) into an adrenal vein for 60 min produced relatively dose- and time-dependent inhibition in CA secretion evoked by ACh (5.32$\times$10$^{-3}$ M), high $K^{+}$ (5.6$\times$10$^{-2}$ M), DMPP (10$^{-4}$ M for 2 min), McN-A-343 (10$^{-4}$ M for 2 min), cyclopiazonic acid (10$^{-5}$ M for 4 min) and Bay-K-8644 (10$^{-5}$ M for 4 min). Furthermore, in adrenal glands pre-loaded with quinine (5$\times$10$^{-5}$ M), CA secretory responses evoked by veratridine (10$^{-4}$ M) was time-dependently inhibited. Also, in the presence of lidocaine (10$^{-4}$ M), which is also known to be a sodium channel blocker, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclo-piazonic acid were also greatly reduced in similar fashion to that of quinidine-treatment. Taken together, these results suggest that quinidine causes greatly the inhibition of CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization, indicating strongly that this effect may be mediated by inhibiting influx of extracellular calcium and release in intracellular calcium in the rat adrenomedullary chromaffin cells. Furthermore, these findings indicate strongly that this inhibitory action of quinidine appears to be associated to the blocking action of sodium channels at least in CA secretion from the rat adrenal gland.and.

• 대한한의학방제학회지
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• 제28권1호
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• pp.41-51
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• 2020

Objectives : US extract is a mixture of each extract of Ulmi cortex and Smilacis rhizoma. In this study, we investigated the antioxidant, anti-inflammatory, antibacterial, and ovoprotective effects of US extract in in vitro model to identify potential candidates for improving female reproductive function. Methods : The antioxidant activity of US extract was measured using 1,1-diphenyl- 2-picrylhydrazyl free radical and superoxide anion radical scavenging assays. The anti-inflammatory effect of US extract on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were determined with a nitric oxide (NO) assay, enzyme linked immunosorbent assays, and western blots analysis. The antibacterial activity of US extract against vaginitis infection microorganisms were determined with disc diffusion and minimum inhibitory concentration assays. The ovoprotective effect of US extract on 4-vinylcyclohexene diepoxide (VCD)-induced ovotoxicity in CHO-K1 cells were evaluated with a cell viability assay. Result : US extract showed good antioxidant capacity and inhibited LPS-induced NO production as well as iNOS and COX-2 expression and secretion of pro-inflammatory cytokine IL-6 without affecting the cell viability. It showed significant clear zones for Staphylococcus aureus and Candida albicans but did not indicate the clear zones for Escherichia coli and Enterococcus faecium. VCD-induced ovotoxicity in CHO-K1 cells was significantly reduced by US extract pre-treatment. Conclusions : These results demonstrate that US extract has antioxidant activity, anti-inflammatory effects on the LPS-stimulated macrophages, antibacterial activity against vaginitis infection microorganisms, and protective effects on the ovarian cells against VCD-induced ovotoxicity. These findings suggest that the US extract can be used as new prescriptions, supplements, functional foods, and cosmetics for improving female reproductive function.

• 대한본초학회지
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• 제29권1호
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• pp.53-60
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• 2014

Objectives : Bojungikgitang (BJT), the Oriental medical prescription has been traditionally used about improvement of immune response and infective disease at Asian nation. In this study, we has compared about the anti-inflammatory and antioxidative effects on BJT of three countries including Korea (Korean Traditional Medicine, KTM), China (Traditional Chinese Medicine, CTM) and Japan (Japanese Traditional Medicine, JTM). Methods : We has basically using LPS-stimulated RAW 264.7 cells. The expression of these inflammatory mediators has measured using enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR). Also, free radical scavenging assay has tested for anti-oxidative activity as well as the contents of total flavonoid and polyphenol. Results : As a result, we were founded the inhibitory effects of BJT (KTM, CTM, JTM) on LPS-induced production of NO, TNF-${\alpha}$ and IL-6 as well as the anti-oxidative activities. Especially the KTM was most effective in anti-inflammatory and anti-oxidative activities. Conclusions : These results indicate that BJT (KTM, CTM, JTM) has a good anti- inflammatory and anti-oxidative effects. But, there were degree of effects on between pharmacopoeia of the countries. Thus, further study is required that find appropriate methods for extracting as well as establish of standardized processes in order to improve the quality of BJT (KTM, CTM, JTM) as an anti-inflammatory and anti-oxidative agent for treatment of inflammatory diseases.

• 대한화장품학회지
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• 제47권1호
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• pp.65-73
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• 2021

피부 염증은 흉터, 노화 뿐만 아니라 아토피와 같은 질환으로 발전할 수 있어 이를 조절하는 것이 매우 중요하다. 본 연구에서는 최근 인체에서 염증을 조절하는 것으로 알려진 specialized pro-resolving mediators (SPMs)의 in vitro 합성과 화장품 적용 가능성을 확인하였다. 대두의 lipoxygenase를 이용하여 mono 또는 di-hydroxy docosahexaenoic acid가 혼합된 시료 S-SPMs를 제작하였고 효능 평가에 이용하였다. 먼저, UVB로 염증을 유도한 세포에서 TNF-α와 IL-6의 발현이 S-SPMs에 의해 감소하고, 미세먼지에 의해 유도된 nitric oxide (NO)의 생성 역시 감소하는 것을 확인하여 S-SPMs의 항염 효능을 확인하였다. 또한, S-SPMs을 처리한 조건에서 malondialdehyde (MDA) 생성이 감소하여 지질 과산화 억제능이 있음을 확인하였고 S-SPMs에 의한 matrix metalloproteinase-1 (MMP-1)의 발현 감소, procollagen type I의 함량 증가를 통해 collagen 분해를 억제하고 반대로 합성은 촉진함을 확인하였다. 끝으로 filaggrin과 loricrin의 발현이 S-SPMs에 의해 증가한 것을 확인하여 피부 장벽 강화 효능을 확인하였다. 위 결과를 토대로 S-SPMs은 피부의 염증 억제와 함께 손상회복, 주름개선 및 장벽 강화를 위한 소재로 활용 가능함을 확인하였다.

• 대한본초학회지
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• 제32권2호
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• pp.77-85
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• 2017

Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

• BMB Reports
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• 제55권6호
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• pp.275-280
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• 2022

The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

• 한국균학회지
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• 제39권1호
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• pp.57-67
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• 2011

신령버섯은 담자균문, 주름버섯목, 주름버섯과에 속하는 식용버섯으로 혈당, 혈압강하효과와 콜레스테롤 저하, 항종양, 암예방 효과가 있는 것으로 알려져 있다. 신령버섯의 자실체로부터 메탄올, 중성염용액 및 열수 등을 이용하여 조다당류를 추출하고 성분을 분석한 결과 ${\beta}$-glucan은 21.54~32.31%, 단백질은 0.16~9.34%로 구성되어 있는 것이 밝혀졌고 신령버섯 추출 조다당류를 Sarcoma 180, HT-29, NIH3T3 및 RAW 264.7 등의 암세포 및 정상세포에 대한 독성을 조사한 결과 10~2000 ${\mu}g/ml$의 조다당류 농도에서 각각의 세포는 세포독성을 나타내지 않았다. Sarcoma 180으로 접종된 ICR 생쥐에 신령버섯의 자실체에서 추출한 각각의 조다당류를 투여한 실험군은 대조군에 비해 수명이 각각 18.8~50.6% 연장되었다. 비장세포의 증식능과 B 임파구의 활성화에 미치는 alkaline phosphatase의 활성을 조사한 결과 비장 세포 증식능은 대조군에 대해 1.1~1.2배의 증식능을 보였고 B 임파구의 활성은 대조군에 대해 1.2~1.6배 증가하였다. 또한 버섯추출 조다당류를 투여한 실험군이 대조군에 비하여 대식세포에서 1.3~4.3배의 많은 nitric oxide를 발생시켰다. 비장세포에 여러 농도의 조다당류를 처리한 후 TNF-${\alpha}$, IL-$1{\beta}$, IL-2 및 IL-6 등의 사이토카인 분비량을 측정한 결과 생성된 사이토카인의 양은 대조군에 비해 2.2배 높게 나타났다. 중성염용액추출 조다당류를 50 mg/kg body weight의 농도로 투여한 실험군 생쥐의 총 복강 세포 수는 대조군에 비하여 4배 증가하였고, 열수추출 조다당류를 50 mg/kg body weight의 농도로 투여한 실험군 생쥐의 백혈구 수는 대조군에 비하여 각각 2.7배 증가하였다. 혈액생화학적 검사를 시행한 결과, 대조군과 유사한 경향을 나타냈다. 따라서 신령버섯의 자실체에서 추출한 조다당류는 생쥐의 면역을 증강시키는 것은 물론 Sarcoma 180에 대한 항암효과를 나타내는 것으로 밝혀졌다.

• 대한한방부인과학회지
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• 제21권3호
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• pp.60-74
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• 2008

Purpose: This study was designed to investigate the effects of Gwibi-Tang(GBT) on the polycystic ovary(PCO) induced by estradiol valerate(EV) in rats. Methods: PCO was induced by single intramuscular injection with EV(4mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for eight weeks. PCO control group(n=8) were injected with EV and orally administrated distilled water for eight weeks. GBT treated group(n=8) were injected with EV and orally administrated GBT for eight weeks. Then we measured weight of body, ovaries, adrenal glands, and uterus of rats. The histopathology changes of ovaries were also evaluated. The expression of nerve growth factor(NGF) was analyzed in the central nervous system, adrenal glands and ovaries by immunohistochemistry. And also CRF expression in median eminance of Rats were analyzed. Results: 1. The weight(g) of rats in GBT treated group($275{\pm}14$) was significantly increased(p<0.01) compared with control group($253{\pm}8$), 2. The weight(mg) of ovaries in GBT treated group($75.8{\pm}16.7$) was significantly increased(p<0.001) compared with control group($37.4{\pm}6.7$). 3. The number of mature follicles in GBT treated group($3.6{\pm}1.2$) was significantly increased(p<0.01) compared with control group($1.5{\pm}1.5$. 4. The number of atretic follicles in GBT treated group($8.0{\pm}3.1$) was significantly decreased(p<0.01) compared with control group($18.6{\pm}6.0$). 5. The number of cystic follicles in GBT treated group($0.5{\pm}0.5$) was significantly increased(p<0.01) compared with control group($2.3{\pm}1.3$). 6. The number of corpora lutea in GBT treated group($6.1{\pm}3.9$) was significantly increased(p<0.01) compared with control group($1.6{\pm}2.3$). 7. The expression of NGF-immunoreactive cells in the ovarian granulosa cells in GBT treated group was lesser observed than control group. Conclusion: From the above results, we concluded that Gwibi-Tang has inhibitory effect on the development of EV-induced polycystic ovary. And it's effect may be related with decreased NGF activities in the ovary.

• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.315-320
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• 2013

Here, we show that radicicol, a fungal antibiotic, resulted in marked inhibition of inducible nitric oxide synthase (iNOS) transcription by the pancreatic beta cell line MIN6N8a in response to cytokine mixture (CM: TNF-${\alpha}$, IFN-${\gamma}$, and IL-$1{\beta}$). Treatment of MIN6N8a cells with radicicol inhibited CM-stimulated activation of NF-${\kappa}B$/Rel, which plays a critical role in iNOS transcription, in a dose-related manner. Nitrite production in the presence of PD98059, a specific inhibitor of the extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) pathway, was dramatically diminished, suggesting that the ERK1/2 pathway is involved in CM-induced iNOS expression. In contrast, SB203580, a specific inhibitor of p38, had no effect on nitrite generation. Collectively, this series of experiments indicates that radicicol inhibits iNOS gene expression by blocking ERK1/2 signaling. Due to the critical role that NO release plays in mediating destruction of pancreatic beta cells, the inhibitory effects of radicicol on iNOS expression suggest that radicicol may represent a useful anti-diabetic activity.