정향 , 마황 , 계피의 간염 B 형 바이러스 증식 억제 효과

강석연(Seog Youn Kang),김태균(Tae Gyun Kim),박민수(Min Su Park),한형미(Hyung Mee Han),정기경(Ki Kyung Jung),강주혜(Ju Hye Kang),문애리(A Ree Moon),김승희(Seung Hee Kim) 한국응용약물학회 1999 Biomolecules & Therapeutics(구 응용약물학회지) Vol.7 No.2

This study was undertaken to test for anti-Hepatitis B virus (HBV) activity of the aqueous extracts prepared from Eugenic caryophyllate, Ephedra sinica, Cinnamomum cassia. Aqueous extracts were assayed for the inhibition of HBV replication by measurement of HBV DNA and surface antigen (HBsAg) levels in the extracellular medium of HepG2 2.2.15 cells. All extracts decreased the levels of extracellular HBV virion DNA at concentrations ranging from 128 to 256 ㎍/㎖ and inhibited the production of HBsAg dose-dependently. Our findings suggest that these three hebal medicinal plants may have potential to develop as specific anti-HBV drugs in the future.

Docosahexaenoic Acid가 Ibotenic Acid 혹은 Scopolamine으로 유발된 흰쥐의 기억력감소에 미치는 영향

강석연(Seog Youn Kang),김승희(Seung Hee Kim),정기경(Ki Kyung Jung),김태균(Tae Gyun Kim),한형미(Hyung-Mee Han),허인회(In Hoe Huh) 한국독성학회 1999 Toxicological Research Vol.15 No.2

The effect of docosahexaenoic acid (DHA) on the memory loss was investigated in two different animal models of dementia (active and passive avoidance models). In the active avoidance retention performance, DHA was administered to experimental animals for 24 days and ibotenic acid, a neurotoxin known to cause destruction of cholinergic nerve terminals, was injected into the nucleus basalis meynert on the 12th day. In the passive avoidance retention performance, DHA was administered to experimental animals for 24 days and either ibotenic acid or scopolamine was injected 24 hours (ibotenic acid) or 30 min (scopolamine) before the experiment. All animals were tested for the active and passive avoidance by the shuttle box and the reflex system, acetylcholinesterase (AChE) activity, AChE distribution by histochemical staining, amyloid protein and synaptophysin distribution by immunohistochemical staining and the degree of lipid peroxidation in the brain. DHA did not have any effect on the inhibition of the active and passive avoidance induced by ibotenic acid. On the other hand, scopolamine-induced inhibition of passive avoidance was partially reversed by DHA, indicating the implication of DHA in potentiation of cholinergic transmission. No differences were observed in AChE activity, AChE distribution, amyloid protein deposition synaptophysin distribution and the degree of lipid peroxidation between DHA-treated and vehicle (10% corn oil)-treated groups. These results demonstrated that administration of DHA for 24 days did not have any effect on the memory loss induced by ibotenic acid whereas scopolamine-induced memory loss was partially reversed by DHA. The precise role of cholinergic transmission potentiating effect of DHA in term of improving memory and/or learning ability remains to be elucidated.

작약감초탕이 미성숙 흰쥐에서 난포성숙 및 Estrogen생성에 미치는 효과

정기경,강석연,김태균,김창옥,문애리,유경자,이송득,류항묵,김승희,Jung, Ki-Kyung,Kang, Seog-Youn,Kim, Tae-Gyun,Kim, Chang-Ok,Moon, A-Ree,Ryu, Kyung-Za,Lee, Song-Deuk,Ryeu, Hang-Mook,Kim, Seung-Hee 한국생약학회 1997 생약학회지 Vol.28 No.3

The traditional herbal medicine, Jackyakgamcho-tang(JGT), was reported to decrease serum testosterone levels and make pregnancy possibel in anovulatory woman and rat. JGT contains Paeoniae Radix(PR) and Glycyrrhizae Radix(GR) in equal amount. This study was designed to investigate the effect of JGT and its components(PR, GR, paeoniflorin and glycyrrhizin) on uterine and ovarian responses, follicular development, and estrogen secretion in the immature rat. The samples(water extracts of JGT, PR, GR; pure compound of paeoniflorin and glycyrrhizin) were administered orally to rats from the 21th day of age to the 28th or 30th days of age for 7 or 9 days. JGT(400mg/kg) and PR(100mg/kg, 200mg/kg) treatments significantly increased serum estradiol above levels in control rats, but both GR and glycyrrhizin had no effect on this parameter. Gross observation and histological analysis revealed that an increased number of growing follicules was observed in the ovaries of JGT and PR treated rat. However the lutenized follicles and ova present in the oviducts were not observed in all rats except one treated with estrogen as a positive control. These results indicate that JGT stimulates the estrogen production and follicular maturation in the immature rat and PR is the main component to induce such reaction.

수종 생약재의 간염 B형 바이러스 증식 억제 활성 검색

김태균,한형미,강석연,정기경,김승희,Kim, Tae-Gyun,Han, Hyung-Mee,Kang, Seog-Youn,Jung, Ki-Kyung,Kim, Seung-Hee 한국생약학회 1999 생약학회지 Vol.30 No.3

This study was undertaken to test for anti-hepatitis B virus (HBV) activity of the aqueous extracts prepared from 9 medicinal plants of Korea (Cornus officinalis, Caesalpinia sappan, Rubus coreanus, Lycium chinense, Artemisia capillaris, Isatis tinctoria, Phyllanthus urinaria, Lysimachia christinae, Lonicera japonica). Aqueous extracts were tested for cytotoxicity and assayed for inhibition of HBV replication by measurement of HBV DNA and surface antigen (HBsAg) levels in the extracellular medium f HepG2 2.2.15 cells. The extract from Rubus coreanus, Artemisia capillaris, Phyllanthus urinaria decreased the levels of extracellular HBV virion DNA at concentrations ranging from 128 to $256\;{\mu}g/ml$ and inhibited the production fo HBsAg dose-dependently without showing cytotoxicity. Our findings suggest that these three hebal medicinal plants may have potential to develop as specific anti-HBV drugs in the future.

새로운 프로톤 펌프 억제제 , IY-81233 의 항위염과 항궤양작용

김승희(Seung Hee Kim),김진(Jeen Kim),강석연(Seog Youn Kang),이송득(Song Deuk Lee),홍성걸(Sung Gul Hong),김동연(Dong Yeun Kim),문애리(A Ree Moon) 한국응용약물학회 1996 Biomolecules & Therapeutics(구 응용약물학회지) Vol.4 No.3

This study was designed to determine the effect of newly synthesized antiulcer agent, 5-pyrrolyl-6-halo-2-(pyridyl-2-methylthio)benzimidazole derivatives (IY-81233), on various experimental ulcers and on the secretion of prostaglandin E² (PGE²) into the gastric lumen of rat. IY-81233 was previously reported to have a strong inhibitory effect on H^+/K^+-ATPase and on gastric acid secretion in rats. Oral administration of IY-81233 at concentrations of 0.2, 2.0, and 20 mg/kg inhibited gastric lesions and duodenal ulcer induced by indomethacin, HCl-ethanol, water-immersion stress, cysteamine, and acetic acid in a dose dependent manner. Their IC_(50) values were 3.4, 1.4, 0.8, 1.3, and 1.2 mg/kg, respectively. These results indicate that IY-81233 is a potent antiulcer agent although it is slightly less potent than omeprazole in healing of gastritis and ulcers. The secretion of PGE² into gastric lumen was also investigated in relation to the cytoprotective effect by IY-81233 in rats. The PGE² level was not changed significantly by an oral administration of IY-81233, suggesting that IY-81233 has little effect on the gastric protection. Therefore, it can be concluded that IY-81233 exerts prominent antiulcer activity by suppressing gastric acid secretion via an inhibition of a proton pump and not by protecting the gastrointestinal mucosa against various ulcerative stimuli.

HepG2 2.2.15 세포주를 이용한 가자, 지유, 복분자, 대황의 B형 간염바이러스 증식 억제 효과

김태균(Tae Gyun Kim),박민수(Min Su Park),한형미(Hyung Mee Han),강석연(Seog Youn Kang),정기경(Ki Kyung Jung),류항묵(Hang Mook Rheu),김승희(Seung Hee Kim) 대한약학회 1999 약학회지 Vol.43 No.4

This study was undertaken to test for antiviral activity of the aqueous extracts prepared from 4 medicinal plants of Korea (Terminalia chebula, Sanguisorba officinalia, Rubus coreanus, Rheum palmatum). Aqueous extracts were assayed for the inhibition of hepatitis B virus (HBV) replication by measurement of HBV DAN and surface antigen (HBsAg) levels in the extracellular medium of HepG2 2.2.15 cells. All extracts decreased the levels of extracellular HBV virion DNA at concentrations ranging from 64 to 128mcg/ml and inhibited the production of HBsAg dose-dependently. Among the 4 tested plants, Terminalia chebula exhibits the most prominent anti-HBV activities. Our findings suggest that these 4 medicinal plants may have potential to develop as specific anti-HBV drugs in the future.

동맥경화질환 동물모델을 이용한 약효검색 연구

김태균,한형미,박민수,강석연,정기경,강주혜,최돈하,김승희,류항묵 식품의약품안전청 1998 식품의약품안전청 연보 Vol.2 No.-

동맥경화증은 미국, 유럽뿐만 아니라 우리나라에서도 죽요 사망원인으로 대두되고 있으며 심혈관계 질환의 대툰분 즉 뇌혈관 질환, 판상동맥 질환,고혈압 및 말초혈관의 홉착 둥이 동맥경화증의 진챙에 의해 발병한다. 본 연구에서는 토끼와 CETP 형질전환 동물모질을 이용하여 느릅나무 수괴의 에탄을 추출물(0.5%), 산마늘의 탄을 추출물(0.5%』과 curcumin(0.1%, 0.5%)의 콜레스테를 저하 및 항동맥경화 효과를 관찰하였다. 토끼 실험결과,느릅나무 수펴 투여군은 혈중 촌 콜레스테롤과 LDL-콜궤스테롤의 농도가 감소 되었으며 간과 심장 조직에서 콜궤스데롤과 TG지질의 함량이 감소히는 것으로 나타났다. 산마늘 투여군에서 혈중 총 콜레스데를 농도는 20.8%(p(0.05) 감소하였교,LDL-콜레스데를 농도의 21-8% (p(0.01) 감소하였다. 간과 심장 조직의 충 걸궤스데를 함량은 각각 45.1% (p(0.05),66.4% (p(0.01) 감소하였다. 0.5% curcumin 투여군은 혈중 총 콜궤스데를 농도, LBL-콜레스데를 농도, 과살화지 질 농도가 유의하게 감소되었고 간파 심장 조직의 TG 함량이 감소되었다. 식험동물의 대동맥을 oil red 0로 염색한 결과, 산마늘과 curcumin 투여군의 대동맥에서 지방반과 아테로마의 형성이 감소되었음이 관찰되었다. CETP 형질 전환 마우스 실험결과,산마늘은 혈중 총 콜레스테를 농도를 9.4% 낮추며 심장의 TG 함량을 낮추는 것으로 나타났다. 본 연구결과에서 느릅나무 수피, 산마늘과 curcumin은 혈액과 조직에서 동맥경화의 병인인 지질과 과산화지질의 농도를 낮추는 갓으로 나타났으며, 산마늘과 curcumin은 동맥의 지질침착을 감소시켰다. 이러한 작용을 나타칙는 약리활성 물질의 꽉인과 기전을 밝히기 위한 좀더 싱도있는 연구가 수쳉되어야 할 것이다. Atheroscler(osis is emerging as one of the majrlr causes of death in Korea as well as Western societies. Most of cardiouascular diseases such as cerebi·ovascular diseases, coronary artery dieeases, hypertension and gangrene result from progress of atherosclerosis. In the present studf,hypocholesteremic and ant:j-atherogenic effect of curcumin(0.1%, 0.5% ) ffmus cravitridna (ethanot eE-tract, 0.5% ) and fffiurt vicforiaff (ethanol extract, 0.5% ) was investigated using conventional rabbitmodel snf cholesteryl ester transfer protein(CETP ) transgenic mouse model. In the experimeot usingthe rabbit model, treatment, with tl daoidiana extract decreased total serum cholesterol and LDL-choles-terol levels as well as tissue chotesterol and triglyceride levels in the heart and the liver. In the rt.oicforiafig-treated group, serum total cholesterol was decreased 13y 20.8% (P <0.05) and LDL-chotesterol by 21.8% (P <0.01). Totat cholesterol contents in the liver and the heart were also decreased by 45.19(P<0.05)』 66.4% (P<0.01), respectively. Treatment with 0.5:4 curcumin significantly decreased the level of total cholesterol,1,DL-cholesterol and malondialdehrdes in the serum and the level of trigtyc☞rides in the liver and the heart. Oil red 0 staining of the aorta isolated from the experimental animals showed that treatment wit'h rt. oiooridfia extract or curcumin decreased fatty streaks aBd atheroma for-mation -ff the aorta. In thr experiment employing CETP transgenic mouse model, f. ufcforiafis eBtractdecreased the level of total eholesterol by 9.4% and the tissue triglyceride levels in the heart. Theseresults demonstrated that V daoidiana and 4. oicforiafis extracts and curcumin decreased the level ofblood serum cholesterol, tissue cholesterol and triglyceride levels, and f. vidoriafis extracts apdcurcumin decreased the cholesteiol accumulation on th,e aorta. The precise mechanism ofhypocholesteremic effect of these plant extracts and curcumin remains to be elucidated.

정소에서 단백질 카르복실메칠화 반응의 기능에 대한 연구 : 돼지 정소로 부터 정제된 protein L-isoaspartyl methyltransferase의 성질에 관한 연구 Characterization of the purified protein L-isoaspartyl methyltransferase from porcine testis

정기영,신미희,한형미,강석연,김태균,홍성렬,김승희,이영근 식품의약품안전청 1997 식품의약품안전청 연보 Vol.1 No.-

Protein carboxylmethyltransferase(PCMT)의 일종인 Protein L-isoaspartyl methyltransferase(PIMt)는노화 혹은 손상된 단백질의 수복에 관여하는 것으로 보고되어 있으며 Rat의 장기별로 활성도를 비교랬을 때 특히 뇌와 정소에서 가장 높은 팔성을 보였다. 그러나 현재까지 정소에서 rIMT의 기능은 명화하게 규명죄어 있지 않으며 다만 포유동물의 정자가 수정전 몇주동안 부고환에 불활섞 상태로 저잖되는례, 이때 PIMT가 sperm protein의 구조 변형을 막음으로써 sperm의 기능과 임신능력에 영향을 미칠 것으로 생각되고 있다- 따라서 향후 PIMT의 기능 손상 과 감성 불임과꼭 상관관계를 밝히기 위해서 일차적으로 래지 정소에서 PIMT를 정제하고 그 특성을 알아보고자 하였다. 퇘지 정소를 도축장에서 구입하여 blender로 균질화한 다음 고속 원심분리와 초고속 원심분리를 이용해서 crtosol fraciion을 얻었다. of c?tosol fraction을 ammonium sulfate로 70% 포화시켜 침전물을 얻은 다음 salt를 제거하기 위해 투석하였디7. 이를 DBAE-serhacel 크로마토그래피, 읖AH-liganded 교finity 크로마토그래피,gel filtration 크로마토그래피를 하섞 SDS-PAG딘로 확인한 결과 순수정제된 PIMT를 얻있으며 분자량은 27, 0009a 정도이고 yield는 1.6%, purification fold는 1550배였다. 정제된 PIMT의 optimum pH는 6,0이었으며 isoelectric pjint(Pl)는 6.2였다. 이효소의 SAM에 대한 K- value는 1.ODf이었고 V-..는 1166.lunits였으며 WP-(L-isoAsp)-HA에 대한 K☞ value는 33.2Dtt이었고 V-..는 741.Ounits 이었으며histone type H-As에 패한 Kf value는 276Df 이었으며 Vma.는 502.Sun죠5 이었다. Protein L-isoaspartyl methyltransferase(PfMT) is Eelieved to 13e play an important role in the disposition of age-rlamaged proteins by catatyzing the repair of abnormal isoaspartyl linkagesresulting flora the spontaneous deamidatioB of asparaginyt resiaues or isomeriBation of aspartylresidues. Although PIMT is widely distributed in maramalian tissues, its activity is especially high intestis. So far, Testicular PIWT could be invoved in either sperm motility or sperm maturation. Thereforeas a initiat step toward caarifying the function of protein carboxfrlmethylation in testis we purifiedPIMT from porcine testis as a monomeric 27,000 Da species by ammoBium sulfate precipftation, DEAE-sephacel chromatography, SAH-liganded affinity chromatography, and gel filtration chromatography.Isoelectric focusing of puril'ied PIMT showed a single protein species with an isoelectric point of 6.2. Theoptimum Bf for the reacti㉠n was 6.0. Km values of the enByme for the S-adenosyl-L- methionine(SAM) ,synthetif'"'ftigopeptide(VYP-L- isoD-HA ) and histone type ll -As were 1.ODf, 33.2ff and 276Dfrespectively.

DHEA의 항비만 효능 및 ob 유전자 (leptin)의 발현에 미치는 영향

정기경,신미희,한형미,강석연,김태균,강주혜,문애리,김승희 德成女子大學校 藥學硏究所 2000 藥學論文誌 Vol.11 No.1

Leptin, the product of the ob gene, is a small peptide molecule synthesized by white adipocytes with an important role in the regulation of body fat and food intake. Baesd on the evidence that synthesis of leptin is regulated by female sex hormone, estrogen, this present study was investigated whether sex hormone precursor, DHEA, can regulate obese gene expression in lean and genetically obese (ob/ob) mice. Antiobesity activity of DHEA was evaluated by determining body weight, food consumption, epididymal fat weight and serum levels of cholesterol and triglyceride in ICR, C57BL/6J, and ob/ob mice. The treatment of C57BL/6J lean and obese mice with a diet containing 0.3% and 0.6% DHEA resulted in lowered rates of weight gain in comparison to non-treated mice, although much greater response was found in the obese mice. All other concentrations of DHEA (0.015%, 0.06%, 0.15%, 0.3%) except the highest one(0.6%) showed no significant effects on weight gain in ICR mice. Food consumption was significantly decreased in all mice treated with 0.6% DHEA, whereas it was not decreased in ICR mice at lower concentrations than 0.6% DHEA. DHEA decreased significantly epididymal adipose tissue weight and serum triglyceride levels dose dependently in lean and obese mice. However, serum cholesterol levels were decreased at lower concentrations than 0.15% DHEA and increased at concentrations of 0.3% and 0.6% DHEA in lean and obese mice. These increases in serum cholestrol levels at high concentrations of DHEA might result from the fact that DHEA has a cholesterol moiety, thereby interfered the assay system. As an approach to elucidate the mechanism for antiobesity activity of DHEA, we examined mRNA levels of obese gene in the adipocyte and obese gene product (leptin) in the serum. The results showed that DHEA did not affect obese gene expression in ICR and C57BL/6J mice. Therefore, we concluded that antiobesity activity of DHEA was not modulated by obese gene expression.

Effects of Curcumin on the Modulation of Microglial Activation(Ⅲ)

Jung, ki Kyung,Lee, Hae Sung,Kim, Tae Gyun,Kang, Ju Hye,Cho, Mi Young,Kim, Seung Hee,Hong, Sungyoul,Kang, Seog Youn 식품의약품안전청 2001 식품의약품안전청 연보 Vol.5 No.-

노인성 치때의 원인물질로 추정되는 Af뀁타이드와 IFN- r로 운도된 microglia의 활성화를 카레의 주성분인 curcumin이 이를 억제하는지 알아보았으며, 또한 도파민에 의해 부분적으로 손상된 신경세포가 활성화된 micro잃.ia회부터 생성된 NO에 의해 손상정도가 증가하는지를 밝히고, curcumin이 NO 캥성을 막제하여 손긍된 신경세포를 보호할 수 있는지를 연구하였다. 생후 0일에서 』일된 rat의 데뇌피질로부터 microgtial cell을 2~3주간 culture하여 얻은 다음 Aβ(25-35)와 IFN-γ로 활성죡를 유도하고 curcLlmin을 2.j~20rH1 범위로 처리했을 패, NO, TNf-o, IL-lr, IL-6의 생성이 용량의존걱으로 억제되었다. 특히 ,4.Ji'(2i-35)와 IFN-r에 의해 활성화된 micro앙la 세포와 도파민에 의해 부분짙으조 손상된 신경세포(CATH.a cell)와의 coculture를 통하여 활성화된 rrllcroglia로부터 생성된 NO가 도과민에 의한 신경세포의 손상을 더욱 더 증가시켰다. 또한 이러한 신경세포의 손상이 J,fl포사멸 (apoptosis)의 형태로 나타남을 밝혔다. 이러한 모델에 curcumin을 적응한 결과 신경세포의 손상이 감소됨을 알 수 있었다. 따라서, 이러한 결과는 curcumin이 Aβ(23-3i)와 IFh- r로 microglia의 확성화를 유도했을 때 생성되는 NO를 억제함으로서 부분적으로 신경J,11포를 보호할 수 있음을 시사한다. Microglia, brain resident macrophages, play a central role in the inflammatory responses of the brain and are activated in brain injuries and several neurodegenerative diseases such as Alzheimer's and Huntigton's disease, thereby aggravating the course of these diseases. In this study we investigated whether curcumin, which is a major component of turmeric (Curcuma longa) and is known as antiinflammatory and antioxidant material, can inhibit production of inflammatory factors in amyloid β(Aβ(25-35)) and interferon-γ-stimulated microglia. Curcumin inhibited production of these factors in a dose dependent manner and also dexamethasone (DEX) inhibited production of them. Continuously, we examined whether curcumin can protect the partially dopamine-damaged neuron through suppression of the production of NO in Aβ(25-35) and interferon-γ-stimulated microglia using potentiation of CATH.a cell death induced by dopamine. Curcumin reduced CATH.a cell death by dopamine level in co-culture with immunostimulated microglia. iNOS inhibitor, NMMA and DEX also reduced CATH.a cell death. Furthermore, both curcumin and NMMA reduced apoptotic death of CATH.a cells in DAPI staining and DNA fragmentation experiments. This result suggest that curcumin can protect neuron in part through suppression of NO produced from Aβ(25-35) plus IFN-γ-stimulated microglia.