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용담사간탕(龍膽瀉肝湯)에 의해 유도된 MAP kinases 활성화를 통한 간암 세포주 HepG2의 세포사멸
윤현정,김한성,허숙경,황성구,박원환,박선동,Yun, Hyun-Jeong,Kim, Han-Seong,Heo, Sook-Kyoung,Hwang, Seong-Goo,Park, Won-Hwan,Park, Sun-Dong 대한한의학방제학회 2007 大韓韓醫學方劑學會誌 Vol.15 No.2
The purpose of this study was to investigate the effect of Yong-dam-sa-gan-tang (YST) on apoptosis in HepG2 cells, First of all. to study the cytotoxic effect of methanol extract of YST on HepG2 cells, the cells were treated with various concentrations of YST and then cell viability was determined by XTT reduction method and trypan blue exclusion assay. YST reduced proliferation of HepG2 cells in a dose-dependent manner. To confirm the induction of apoptosis, HepG2 cells were treated with various concentrations of YST. The cleavage of poly AD P-ribose polymerase (P ARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of caspase-3, procaspase-8 and procaspase-8 were examined by western blot analysis. YST decreased procaspase-3, procaspase-8 and procaspase-9 levels in a dose-dependent manner and induced the clevage of PARP. YST triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome c from mitochondria to cytosol. Furthermore, YST also downregulated the anti-apoptotic Bcl-2 and upregulated the pro-apoptotic-Bax. Therefore, this result suggest that YST induced HepG2 cell death through the mitochondrial pathway. Sustained activation of the Ras/Raf/MEK/ERK cascade in cells results in a cell cycle arrest and has been implicated in the differentiation of certain cell types, in many cases acting to promote differentiation. YST decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. These results suggest that YST is potentially useful as a chemo-therapeutic agent in HepG2.
마우스 대식세포에서 천련자(川련子)의 항산화 및 항염증 효과
이효승 ( Hyo Seung Yi ),허숙경 ( Sook Kyoung Heo ),윤현정 ( Hyun Jeong Yun ),김병완 ( Byung Wan Kim ),박선동 ( Sun Dong Park ) 대한본초학회 2008 大韓本草學會誌 Vol.23 No.4
Objectives: Melia toosendan (MT) has been used as a traditional Korean herbal medicine, and today it is used as a medication for colic, side aches, heartache and other disorders of liver. The aim of this study was to determine whether fractionated extracts of MT inhibit free radical generation such as DPPH radical, superoxide radical and nitric oxide, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods: MT extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of MT onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results: Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H2O2, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-α, IL-1β and IL-6 formation in macrophages. Conclusions: These results indicate that dichioromethane and ethyl acetate extracts of MT have potential as an agent of chronic inflammatory diseases.
사람 대동맥 평활근 세포에서 이묘산(二妙散)의 항동맥경화 활성
윤현정 ( Hyun Jeong Yun ),허숙경 ( Sook Kyoung Heo ),이효승 ( Hyo Seung Yi ),김태훈 ( Tae Hoon Kim ),김동완 ( Dong Wan Kim ),김선모 ( Sun Mo Kim ),박선동 ( Sun Dong Park ) 대한본초학회 2008 大韓本草學會誌 Vol.23 No.4
Objectives: This study was evaluated to elucidate the inhibitory potential of Imyosan (IMS) and its components, Phellodendri Cortex (PC; Phellodendron amurense Rupr., Hwangbaek in Korean) and Atractylodis Rhizoma (AR; Atratylodes lancea D.C., Changchool in Korean), on human aortic smooth muscle cells (HASMC) migration and production of matrix metalloproteinase (MMP)-2 and MMP-9 by TNF-α treatment. Methods: Cytotoxic activity of IMS and its components on HASMC was using 5-(3-caroboxy meth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Effect of IMS, PC and AR on TNF-α-induced HASMC migration underside of matrigel filter was stained with hematoxylin-eosin. And total number of cells that migrated to the underside of the filter was counted. MMP-2 and MMP-9 activity was evaluated by gelatin zymography assay. Results: The matrigel migration assay showed that IMS effectively inhibited the TNF-α-induced migration of HASMC. Moreover, IMS significantly inhibited MMP-9 activity. Our present study demonstrates that IMS and its components inhibit TNF-α-induced HASMC migration and MMP-9 activity. The inhibitory effect of IMS extract is more potent than that of its component herb extracts. Conclusions: These results provide evidence that IMS has multiple effects in the inhibition of HASMC migration and may offer a therapeutic approach to block HASMC migration.
김판준 ( Pan Joon Kim ),윤현정 ( Hyun Jeong Yun ),허숙경 ( Sook Kyoung Heo ),김경애 ( Kyoung Ae Kim ),김동완 ( Dong Wan Kim ),김재은 ( Jae Eun Kim ),박선동 ( Sun Dong Park ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.2
Objectives: Inflammation is important event in the development of vascular diseases including hypertension, atherosclerosis, and restenosis. Bodusan (BDS) was a traditional Korean herbal medicine and widely used in treatment of gastrointestinal complaint and stomach ulcer. The aim of this study was to determine whether BDS and its components inhibit production of nitrite, PGE2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods: Cytotoxic activity of BDS and its components on RAW 264.7 cells was using 5-(3-caroboxymcrophages.eth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results: Our results indicated that BDS and its components significantly inhibited the LPS-induced NO and PGE2 production. Moreover, BDS and its components inhibited iNOS and COX-2 expression accompanied by an attenuation of TNF-α, IL-1β, IL-6 and MCP-1 formation in macrophages. Conclusions: These results indicate that BDS and its components have potential as an anti-inflammatory agent.
혈관내피세포에서 토복령(土茯령)의 항산화 및 항염증 효과
이창현 ( Chang Hyun Lee ),이효승 ( Hyo Seung Yi ),김재은 ( Jae Eun Kim ),허숙경 ( Sook Kyoung Heo ),차창민 ( Chang Min Cha ),원찬욱 ( Chan Wook Won ),박선동 ( Sun Dong Park ) 대한본초학회 2009 大韓本草學會誌 Vol.24 No.3
Objectives: Smilacis glabrae rhizoma (SG) has been traditionally used as a herbal medication of musculoskeletal disorders like arthritis, pain, convulsions, and syphilis in traditional Korean medicine. This study was investigated anti-oxidative and anti-inflammatory effect of fractionated extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (HUVEC). Methods: SG extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of SG onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of ICAM-1 and VCAM-1 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results: Our results indicated that fractionated extracts, especially ethyl acetate (EA) extract, significantly inhibited free radical generation, the TNF-α-induced intracellular oxidation. Furthermore, the EA extract protected TNF- α-induced adhesion to THP-1, expression of adhesion molecules accompanied by an attenuation of IL-6 and IL-8 formation in HUVEC. Conclusions: These results indicate that EA extract of SG have potential as an agent of atherosclerosis and other chronic inflammatory diseases including diabetes, hypertension, and arthritis.