• 제목/요약/키워드: Ca-protein binding

검색결과 245건 처리시간 0.02초

가미청폐사간탕(加味淸肺瀉肝湯)이 고지방식이 비만생쥐의 체지방에 미치는 영향 (Effects of Gami-Cheongpyesagan-Tang on Body Fat in High Fat Diet-Fed Obese Mice)

  • 금선오;이하일;이종하;윤용일;권영미;송용선
    • 한방비만학회지
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    • 제15권2호
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    • pp.75-92
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    • 2015
  • Objectives: This study was designed to investigate the effect of Gami-cheongpyesagan-tang extract (GCST) on high fat diet-induced obesity in rats. Methods: The mice were divided into six groups; normal diet control, high fat diet control (HFD), HFD+GCST administrated group (100, 200, and 400 mg/kg) and olistat-admistrated group. Obesity was induced by high fat diet (45%) for 7 weeks in mice, and GCST was administrated orally every day for 7 weeks. The body weight, food intake, and serological markers such as total cholesterol, triglyceride, lipid contents, leptin, adiponectin and glutamic oxaloacetic transaminase/glutamic pyruvic transaminase were measured in mice. The mRNA expression of obese-associating genes such as sterol regulatory element-binding protein (SREBP)-1c, fatty acid synthase (FAS), stearoyl-CaP desaturase (SCD-1), peroxisome proliferator-activated receptor $(PPAR)-{\alpha}$, COA oxidase (ACO), and carnitine palmitoyltransferase ($CPT-1{\alpha}$) was analyzed by reverse transcription polymerase chain reaction. Results: The administration of GCST at 400 mg/kg, significantly reduced the increase of body weight and food intake as well as food efficiency compared to HFD group. GCST decreased the serum levels of triglyceride, total cholesterol, low-density lipoprotein-cholesterol, leptin in HFD control group and inhibited lipid accumulation in liver and adipose tissues, but did not increase high-density lipoprotein-cholesterol. In the liver tissues of GCST administrated HFD group, the mRNA levels of SREBP-1c, FAS and SCD-1 were decreased and the mRNA levels of $PPAR-{\alpha}$, ACO, and $CPT-1{\alpha}$ were increased. Conclusions: These results indicate that GCST could improve high fat diet induced obesity through inhibiting the hyperlipidemia in fatty Liver. It suggest that GCST may be used clinically for declining the accumultion of body fat with hyperlipidemia.

남극삿갓조개 (Nacella concinna) 장의 중금속 축적에 관한 연구 (Bioaccumulation of Heavy Metals in Intestine of Nacella concinna)

  • 이용석;조용훈;한연수;고원규;안인영;정계헌
    • 한국패류학회지
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    • 제22권1호
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    • pp.87-95
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    • 2006
  • 본 연구에서는 남극삿갓조개 (Nacella concinna) 의 장 (intestine)의 중금속 농축에 관하여 알아보고자 카드뮴 노출실험을 실시한 후 중금속에 노출 시 동물체내에서 유도되는 단백질 중의 하나인 metallothionein을 면역조직화학적 방법으로 추적하고 투과전자현미경 (transmission electron microscope) 을 이용하여 세포의 미세구조의 변화를 알아보았다. 더불어 SEM-EDS 장비를 이용한 원소분석을 통해 중금속의 분포를 알아보아 중금속의 축적 및 해독기작에 대한 기초자료를 얻고자 수행되었다. Metallothionein의 분포를 살펴보기 위하여 면역조직화학적 실험이 수행된 바, 중금속이 농축되고있는 삿갓조개의 장상피세포의 첨단부에 metallothionein이 많이 존재하고 있음을 알 수 있었으며, 미세구조 관찰결과 노출시간이 경과함에 따라 핵막의 팽창, whorl 구조의 출현, 핵 내 봉입체를 관찰할 수 있었다. SEM-EDS 관찰결과 카드뮴 노출결과, 황이 급격하게 줄어들었고, 칼슘과 아연이 상대적으로 증가하는 양상을 관찰 할 수 있었다. 이상에서와 같이 Cd 노출의 정도에 따라 비교적 빠른 세포반응을 보이는 남극삿갓조개의 특징은 자연 상태에서 Cd의 노출에 따른 효과적인 생물 지표종으로서 가치가 있는 것으로 사료된다.

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수도묘(水稻苗)의 Butachlor 흡수(吸收) 및 약해발생(藥害發生) 특성(特性)에 관한 생리적(生理的) 연구(硏究) (Uptake of Butachlor by Rice Seedlings and Its Phytotoxic Action to the Physiological Activities)

  • 정봉진;권용웅
    • 한국잡초학회지
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    • 제1권1호
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    • pp.57-68
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    • 1981
  • 수도묘(水稻苗)의 Butachlor(2-chloro-2',6'-diethyl-N-(carboxymethyl) acetanilide) 흡수특성(吸收特性)과 약해발생가구(藥害發生機構)를 밝히어 Butachlor의 안전사용(安全使用)에 도움을 주고자 시도(試圖)되었다. 수도품종(水稻品種) '만석(萬石)'(수원(水原)264호(號))을 사용(使用)하여 제(第)6, 7여기(葉期)까지 수경재배(水耕栽培)한 후 Butachlor를 0, 1.8, 3.6, 7.2, 10.8 또는 14.4ppm 되도록 처리(處理한) 수경액(水耕液)으로 1, 2, 4일간(日間) 생육(生育)시킨 경우와 Butachlor 처리(處理) 후(後) 정상수경액(正常水耕液)으로 교체(交替)하고 6일간(間生) 생장(生長)시킬 경우 약해(藥害)의 진전(進展) 또는 회복과정중(恢復過程中)에 일어나는 수도(水稻)의 Butachlor 흡수(吸收)에 따른 생장반응(生長反應), 수분흡수(水分吸收), 양분흡수(養分吸收), 기공(氣孔) 개폐(開閉), 질산환원효소(窒酸還元酵素)(Nitrate reductase)의 생합성(生合成) 및 분해작용(分解作用)의 변화(變化)를 검정(檢定)했고 Butachlor의 세포구성물질(細胞構成物質)들에 대한 흡착특성(吸着特性)을 비교(比較)하였으며 그 결과(結結)는 다음과 같이 요약(要約)된다. 1. 수도묘(水稻苗)의 Butachlor의 흡수(吸收)는 처리농도(處理濃度) 및 기간(期間)에 비례(比例)하여 거의 직선적(直線的)으로 증가(增加)하였다. 2. Butachlor는 지상부(地上部) 생육(生育)보다는 뿌리생육(生育)을, 지상부(地上部) 생체중(生體重) 및 출엽(出葉)보다는 초장(草長)의 생육(生育)을 더욱 저해(沮害)하였으며, 처리종료(處理終了) 후(後) 지상부(地上部) 생체중(生體重)과 출엽(出葉)은 조속(早速)히 회복(恢復)되었으나 초장(草長) 및 뿌리생체중(生體重) 생장(生長)은 4일(日) 이후(以後) 회복세(恢復勢)를 보였다. 3. Butachlor는 뿌리의 수분흡수(水分吸收)를 처리농도(處理濃度) 비례(比例)하여 곧 저해(沮害)하였고, 그 결과(結果) 엽면(葉面) 기공(氣孔)의 저항(低抗)을 증가(增加)시켰으며, 처리종료(處理終了) 후(後)에는 수분흡수력(水分吸收力)은 곧 회복(恢復)되었으나 기공(氣孔)의 개도(開度)는 서서히 회복(恢復)되었다. 4. Butachlor는 처리(處理) 전(前) 후(後) 수도묘(水稻苗)의 $NH_4^+$, $K^+$$Ca^{++}$ 같은 양(陽)이온의 흡수(吸收)에는 영향(影響)하지 않았으나 $NO_3^-$의 흡수(吸收)를 뚜렷이 저해(沮害)했고, 7.2 ppm의 고종도(高濃度)에서는 인산(燐酸)의 흡수(吸收)도 저해(沮害)했다.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2016년도 춘계학술대회 및 임시총회
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Kainic Acid로 처리한 해마박편배양 마우스 간질모델에서 치아이랑 Parvalbumin 면역 반응성 사이신경세포의 형태학적 변화 (The Morphologic Changes of Parvalbumin- Immunoreactive Interneurons of the Dentate Gyrus in Kainate-Treated Mouse Hippocampal Slice Culture Epilepsy Model)

  • 정희선;신미영;김영훈;이인구;황경태;김명석
    • Clinical and Experimental Pediatrics
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    • 제45권12호
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    • pp.1551-1558
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    • 2002
  • 목 적 : 중증 측두엽간질환자와 kainic acid(KA)로 처리한 측두엽간질 동물모델에서 해마 치아이랑의 사이신경세포가 소실된다. 측두엽간질 마우스모델인 마우스해마의 기관형 배양에 의한 KA 간질모델에서 parvalbumin(PV)항체를 이용한 면역조직화학법으로 치아이랑에 분포하는 PV 면역반응성 사이신경세포를 감별염색하여 세포체 및 그 가지돌기의 형태학적 변화를 관찰함으로써 측두엽간질의 병태생리의 일단을 규명하고자 한다. 방 법 : 실험적 간질 모델은 C57/BL6 마우스의 해마박편을 이용한 기관형 배양에서 $10{\mu}M$ KA 투여로 유발시켰으며, PV 항체를 이용한 광학현미경적 면역조직화학법으로 치아이랑에 분포하는 PV 면역반응성 사이신경세포를 감별염색하여 형태학적 차이를 관찰하였고, 또한 이들 세포의 세포수를 계측하고 KA처리 후 배양하면서 시간경과(8, 24, 48, 72시간)에 따라 PV 면역반응성 사이신경세포의 형태학적 변화를 관찰하였다. 결 과: KA 처리를 하지 않고 배양된 해마조직박편(대조군)의 치아이랑에서 PV 면역반응성 세포는 가지돌기 나무가 잘 발달되어 있고 사이신경세포로서 이들은 주로 과립층과 이층 밑의 다형층에 산재하였다. $10{\mu}M$ KA에 1시간 정도 노출되었을 때 PV 면역반응성 사이신경세포의 돌기에는 염주상이 형성되었고 가지돌기는 가늘어져 있었다. PV 면역반응성 사이신경세포는 KA 처리 후 배양액에서 KA를 제거한 뒤 시간이 경과함에 따라 형태학적 회복을 보여주었다. KA 제거 후 8시간 회복군에서 세포돌기의 염주상은 볼 수 없었으며 가지돌기는 가늘어져 있었다. KA 제거 후 24, 48, 72시간 회복군에서도 염주상은 거의 볼 수가 없었으며 가지돌기의 두께도 회복되었다. PV 면역반응성 사이신경세포의 수는 대조군에 비하여 KA 처리군과 KA 제거 후 8시간 회복군에서는 통계학적으로 유의한 세포수의 감소가 있었으나 KA 제거 후 24시간 회복군, KA 제거 후 48시간 회복군 및 KA 제거 후 72시간 회복군에서는 대조군과 차이가 없 었다. 결 론: 이러한 결과는 KA에 의하여 유발된 치아이랑 사이신경세포의 세포소실이 일시적이며 가역적인 현상임을 말해주는 것이다.