• 대한한방부인과학회지
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• 제24권2호
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• pp.13-32
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• 2011

Objectives: This study was performed to elucidate the effects of Danchisoyo-san (DS) on cell damage and gene expression in UVB-exposed dermal fibroblast. Methods: To demonstrate the inhibitory effects of DS on aging of the skin, we used human dermal fibroblast(F6) and UVB light(30 mJ/$cm^2$) was used to damage to dermal fibroblast. We measured the nitrite production, LDH release, and gene expression in UVB-irradiated dermal fibroblast to elucidate the actionmechanism of DS. Also, we evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit, and gene expression (MMP-1, procollagen, c-fos, c-jun, NF-kB, Bcl-2, Bcl-xL, iNOS) were determined using real-time PCR. Results: 1. DS inhibited LDH-release, nitrite production in UVB-irradiated dermal fibroblast. 2. DS suppressed the gene expression of MMP-1 in UVB-irradiated dermal fibroblast. 3. DS increased the gene expression of procollagen in UVB-iradiated dermal fibroblast. 4. DS suppressed the gene expression of c-jun, c-fos, NF-kB, iNOS in UVBirradiated dermal fibroblast. 5. DS increased the gene expression of Bcl-2 in UVB-iradiated dermal fibroblast. 6. DS increased the cell proliferation of dermal fibroblast. Conclusions: From the results, we concluded DS increases the cell proliferation and collagen synthesis in dermal fibroblast. So we suggest that DS has the antiwrinkle effects.

• The Korean Journal of Physiology and Pharmacology
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• 제9권2호
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• pp.117-123
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• 2005

A cumulative evidence indicates that consumption of tea catechin, flavan-3-ol derived from green tea leaves, lowers the risk of cardiovascular diseases. However, a precise mechanism for this cardiovascular action has not yet been fully understood. In the present study, we investigated the effects of different green tea catechins, such as epigallocatechin-3 gallate (EGCG), epigallocatechin (EGC), epicatechin-3 gallate (ECG), and epicatechin (EC), on angiotensin II (Ang II)-induced hypertrophy in primary cultured rat aortic vascular smooth muscle cell (VSMC). [$^3H$]-leucine incorporation was used to assess VSMC hypertrophy, protein kinase assay, and western blot analysis were used to assess mitogen-activated protein kinase (MAPK) activity, and RT-PCR was used to assess c-jun or c-fos transcription. Ang II increased [$^3H$]-leucine incorporation into VSMC. However, EGCG and ECG, but not EGC or EC, inhibited [$^3H$]-leucine incorporation increased by Ang II. Ang II increased phosphorylation of c-Jun, extracellular-signal regulated kinase (ERK) 1/2 and p38 MAPK in VSMC, however, EGCG and ECG , but not EGC or EC, attenuated c-Jun phosphorylation increased by Ang II. ERK 1/2 and p38 MAPK phosphorylation induced by Ang II were not affected by any catechins. Ang II increased c-jun and c-fos mRNA expression in VSMC, however, EGCG inhibited c-jun but not c-fos mRNA expression induced by Ang II. ECG, EGC and EC did not affect c-jun or c-fos mRNA expression induced by Ang II. Our findings indicate that the galloyl group in the position 3 of the catechin structure of EGCG or ECG is essential for inhibiting VSMC hypertrophy induced by Ang II via the specific inhibition of JNK signaling pathway, which may explain the beneficial effects of green tea catechin on the pathogenesis of cardiovascular diseases observed in several epidemiological studies.

• 약학회지
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• 제41권5호
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• pp.629-637
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• 1997

Tumor necrosis factor-${alpha}$ (TNF) induced a cytotoxic response in ME-180 cervical carcinoma cells in vitro. This cytotoxic response was accompanied by a temporal series of mitogenic stimuli : increased c-fos, c-jun and jun-B expression. Depletion of protein kinase C (PKC) by exposure of ME-180 cells to 100ng/ml phorbol myristate acetate (PMA) for 24hours almost completely abolished TNF-mediated increase in these signals, indicating that a PKC-dependent pathway is involved in TNF-mediated increases in the expression of c-fos, c-jun and jun-B. Characteristics of TNF receptors after exposure to 100ng/ml PMA or 24hours were not altered, suggesting that diminished induction of these oncogenes by TNF after PMA treatment is not due to any changes at the receptor level. To examine whether a PKC-dependent pathway is involved in TNF-mediated cytotoxicity in ME-180 cells, cytotoxicity was measured after depletion of PKC. No apparent changes in cytototoxicity after PKC depletion suggest that a PKC-dependent pathway is not involved in TNF-mediated cytotoxicity. Furthermore, results from cytotoxicity tests after exposure to staurosporine (PKC inhibitor) did not show any changes in the TNF-mediated cytotoxicity, confirming that a PKC-dependent pathway is not involved in this process. These data indicate that 1) TNF induces expression of c-fos, c-jun and jun-B oncogenes via a PKC-dependent pathway and 2) PKC-dependent expression of these three oncogenes by TNF may not be involved in TNF-mediated cytotoxicity in ME-180 cells.

• 동의신경정신과학회지
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• 제14권1호
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• pp.1-16
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• 2003

Objective : This study was designed to assess the protective effects of Hwangryeonhaedoktang on the animal model of depression, induced by chronic mild stress(CMS). Method : Male Sprague-Dawley rats were used for this experiment. The subjects were divided into 3 groups ( 1. CMS-drug: Hwangryeonhaedoktang administered during CMS treatment, 2. CMS-vehicle: water administered, 3. normal ). After 4 weeks of CMS treatment, they were executed forced swimming test(FST), open field test and c-Fos in paraventricular nucleus(PVN) were measured. Result : 1. In FST, immobility behavior decreased significantly in CMS-drug group. 2. There was no difference in the open field test between 3 groups 3. c-Fos expressed cell bodies in PVN were significantly less in CMS-drug than in CMS-vehicle group. Conclusion : These results suggest that Hwangryeonhaedoktang may have protective antidepressant effects in CMS model rats. And these effects could be explained by the elevated stress-copying behaviors which are related with PVN of hypothalamus.

• Journal of Microbiology and Biotechnology
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• 제18권10호
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• pp.1641-1647
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• 2008

Amygdalin is a cyanogenic glycoside plant compound found in the seeds of rosaceous stone fruits. We evaluated the anti-inflammatory and analgesic activities of amygdalin, using an in vitro lipopolysaccharide (LPS)-induced cell line and a rat model with carrageenan-induced ankle arthritis. One mM amygdalin significantly inhibited the expression of TNF-$\alpha$ and IL-l$\beta$ mRNAs in LPS-treated RAW 264.7 cells. Amygdalin (0.005, 0.05, and 0.1 mg/kg) was intramuscularly injected immediately after the induction of carrageenan-induced arthritic pain in rats, and the anti-arthritic effect of amygdalin was assessed by measuring the weight distribution ratio of the bearing forces of both feet and the ankle circumference, and by analyzing the expression levels of three molecular markers of pain and inflammation (c-Fos, TNF-$\alpha$, and IL-l$\beta$) in the spinal cord. The hyperalgesia of the arthritic ankle was alleviated most significantly by the injection of 0.005 mg/kg amygdalin. At this dosage, the expressions of c-Fos, TNF-$\alpha$, and IL-l$\beta$ in the spinal cord were significantly inhibited. However, at dosage greater than 0.005 mg/kg, the pain-relieving effect of amygdalin was not observed. Thus, amygdalin treatment effectively alleviated responses to LPS-treatment in RAW 264.7 cells and carrageenan-induced arthritis in rats, and may serve as an analgesic for relieving inflammatory pain.

• The Korean Journal of Pain
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• 제26권3호
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• pp.255-264
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• 2013

Background: We investigated the effects of pre-emptive administration of ketamine and norBNI on pain behavior and the expression of DREAM, c-Fos, and prodynorphin proteins on the ipsilateral side of the rat spinal cord at 2 and 4 hours after formalin injection. Methods: Eighty-four male Sprague Dawley rats were divided into 4 major groups consisting of control rats (C) (n = 12), rats given only formalin injections (F) (n = 24), and rats treated with pre-emptive administration of either ketamine (K+F) (n = 24) or norBNI (N+F) (n = 24). The non-control groups were further divided into subgroups consisting of rats that were sacrificed at 2 and 4 hours (n = 12 for each group) after formalin injection. Pain behavior was recorded for 1 hour. After 2 and 4 hours, the rats were sacrificed and the spinal cords (L4-L5 sections) were removed for immunohistochemistry and Western blot analysis. Results: The pain behavior response was reduced in the K+F group compared to the other groups during the second phase of the formalin pain response. We detected an increase in the nuclear DREAM protein level in the K+F group at 2 and 4 hours and a transient decrease in the N+F group at 2 hours; however, it increased at 4 hours after injection. Fos-like immunoreactivity (FLI) and Prodynorphin-like immunoreactivity (PLI) neurons decreased in the K+F group but increased in the N+F group at 2 hours after injection. While FLI decreased, PLI increased in all groups at 4 hours after injection. Conclusions: We suggest that NMDA and kappa opioid receptors can modulate DREAM protein expression, which can affect pain behavior and protein transcriptional processes at 2 hours and bring about either harmful or protective effects at 4 hours after formalin injection.

• 대한의생명과학회지
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• 제11권3호
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• pp.319-326
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• 2005

It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.

• Korean Journal of Acupuncture
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• 제36권3호
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• pp.162-170
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• 2019

Objectives : Increasing evidence suggests that parasympathetic vagus nerve activity plays a role in modulating acupuncture-induced anti-inflammatory reaction, but the function of sympathetic nerve is not known. Here, we investigated whether splanchnic sympathetic nerve activity was involved in the regulation of splenic expression of $TNF-{\alpha}$ mRNA by electroacupuncture (EA) in LPS-injected animals. Methods : DiI was injected into the stomach or celiac ganglion (CG) for retrograde labeling of the target tissues. EA was given at ST36 and the electrical stimulation on the sciatic nerve in LPS-injected mice. c-Fos signals in the tissues were analyzed by immunofluorescence staining, and $TNF-{\alpha}$ mRNA was analyzed by real-time PCR. Results : Application of EA at ST36 or electrical stimulation on the sciatic nerve induced c-Fos expression in neurons of the spinal cord and celiac ganglion (CG). Then, the vagotomy reduced c-Fos levels in CG neurons but not in the spinal cord in animals given EA. Expression of $TNF-{\alpha}$ mRNA which was induced in the spleen after LPS was significantly inhibited by EA, then the vagotomy elevated $TNF-{\alpha}$ mRNA level similar to that in LPS-injected animals. Splanchnectomy in animals given LPS and EA also increased $TNF-{\alpha}$ mRNA though it was less effective than vagotomy. Conclusions : Our data suggest that EA delivered to the spleen via the splanchnic sympathetic nerve may be involved in attenuating splenic inflammatory responses in LPS-injected animals.

• Korean Journal of Acupuncture
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• 제36권2호
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• pp.115-126
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• 2019

Objectives : Osteoporosis is a condition characterized by low bone mass and increased bone fragility. It has become a major problem of senior citizens. The purpose of this study is to experiment the effect of water extract of Forsythiae Fructus (wFF) on osteoclast differentiation; and the other purpose is to examine the effect of wFF on osteoporosis in ovariectomized rat. Methods : To investigate the effect of wFF on osteoclast differentiation and activity, RAW 264.7 cells were used. The number of TRAP positive cell, TRAP activity, pit area, mRNA expression of makers (RANK, TRAP, CA II, CTK, MMP-9, NFATc1, c-Fos), protein expression of makers (NFATc1, c-Fos) were investigated. For in vivo study, 40 female Sprague-Dawley (SD) rats were induced osteoporosis by ovariectomy (OVX) and then tested for anti-osteoporosis effect by administration of wFF. Results : wFF suppressed osteoclatogenesis, TRAP activity and pit area formation. Moreover, wFF decreased the expression of master differentiation factors (NFATc1, c-Fos) and also reduced the osteoclastogenesis-related markers (TRAP, CA II, CTK, MMP-9). These suggest that wFF inhibit osteoclasts differentiation and bone resorption. In the OVX rat model, wFF inhibited decreasing of BMD and trabecular area. Conclusions : Forsythiae Fructus should be effective for osteoporosis prevention and treatment.

• 생명과학회지
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• 제24권5호
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• pp.567-572
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• 2014

초롱꽃과에 속하는 도라지는 항염증, 항산화, 항암작용뿐 아니라 간기능 강화에도 작용하여 한방에서 만성염증성 질환의 치료제로 널리 사용되고 있다. 이번 연구에서 마우스 대식세포주인 RAW 246.7 세포와 비장으로부터 분리한 T세포에서 도라지 추출액(CC), 도라지 추출물이 함유된 활맥단 추출액(HWAL, 도라지 82% 함유) 및 맥환추출액(MAEK, 도라지 7% 함유)을 처리하여 대식세포와 T세포의 활성과 관련 있는 유전자의 발현을 통해 면역증진 및 항염증 효과를 real-time RT-PCR 기법을 통해 확인해 보았다. 그 결과, 활성화된 T세포에서 많이 발현되는 c-fos (10배 이상), CD40L 유전자(6배 이상)의 경우, 맥환 추출액 처리군에서 가장 높은 발현이 유도되어 면역활성 효과가 높을 것으로 사료되었다. 도라지 추출액과 활맥단 추출액의 경우도 c-fos 유전자의 발현을 유의적으로 증가시켰지만 미약하였고, CD40L의 경우는 영향을 주지 않았다. 도라지 추출액을 처리한 군에서 면역활성을 유도하는 M1 대식세포에서 많이 발현되는 iNOS 유전자의 발현이 유의하게 증가되었다. 반면에 활맥단 추출액을 처리한 대식세포는 M2 표식인자인 Ym1, arginase1 (ARG1) 유전자의 발현이 유의적으로 상승하였다. 결론적으로 도라지가 다량 포함된 맥환 추출액은 면역활성을 유도하는데 탁월한 효과를 나타내며, 도라지 함유가 적게 포함된 활맥단 추출액은 항염증 작용에 탁월한 효과가 있을 것으로 사료된다.