황금이 백서의 간 조직 글루타치온 에스-전이 효소 Yc1/2의 발현 효과

김영숙(Young Sook Kim),김동현(Dong Hyun Kim),최미정(Mi Jung Choi),김성민(Sung Min Kim),박래길(Rae Kil Park),권강범(Kang Beom Kwon),류도곤(Do Gon Ryu),김복량(Bok Ryang Kim) 한의병리학회 2003 동의생리병리학회지 Vol.17 No.6

加味馬錢五子丸 추출물이 雄性 白鼠의 성기능에 미치는 효과

권강범(Kang Beom Kwon),김복량(Bok Ryang Kim),황인진(In Jin Hwang),김은경(Eun Kyung Kim),김구환(Goo Hwan Kim),고광학(Kwang Hak Ko),최연성(Yeon Seong Choi),장근국(Geun Gook Zhang),노성일(Seong Il Rho),한동원(Dong Won Han),차숙(Suk Cha 한의병리학회 2004 동의생리병리학회지 Vol.18 No.5

左歸飮 煎湯液이 雄性 白鼠의 性行動 性호르몬에 미치는影響

최기방(Choi, Ki?Bang),박준수(Park, Jun-Su),권강범(Kwon, Kang-Beom),이호섭(Lee, Ho-Sub),박관하(Park, Kwan-Ha),김복량(Kim, Bok-Ryang),류도곤(Ryu, Do-Gon) 대한한의학회 1999 대한한의학회지 Vol.20 No.1

가물치 면역단백질에 관한 硏究

金福亮,鄭憲鐸,白汶基 圓光大學校 醫科學硏究所 1985 圓光醫科學 Vol.1 No.1

In atempt to analyze the molecular weight, structure, and subunit components of serum immunoglobulin from snakehead, purification was done by serial procedures ; sodium sulfate fractionation, DEAE-cellulose chromatography, Sephadex G-200 gel filtration, and Sephadex G-200 gel refiltration. Immunoglobulin titer was detected by the hemagglutination method. The purified snakehead immnuglobulin was shown a symmetric peak when it was eluted through a Sephadex G-200 containing column and shown a single major band by SDS/3% polyacrylamide-0.5% agarose composite gel electrnphoresis. The following results were obtained ; 1. The molecular weight of the purified immunoglobulin is about 700,000. 2. The purified immunoglobulin is organized a tetrameric structure of 7 S subunits. 3. The molecular weight of the H-and L-chains are 70,000 and 22,000 resoectively.

拘杞雙和湯 煎湯液이 雄性 白鼠의 性行動 및 性호르몬에 미치는 影響

金容福,李昊燮,朴俊秀,朴寬夏,柳道坤,權康範,金福亮 대한동의병리학회 1999 동의생리병리학회지 Vol.13 No.1

本 實驗은 성기능 억제제에 의해서 低下되는 男性의 性機能(혹은 生殖機能)을 枸杞雙和湯을 利用하여 保護할 수 있는 實驗을 수행하였다. 男性의 性機能을 低下시키는 物質로는 環境汚染物인 2,3,7,8-Tetrachlorodibenzo-p-dioxin(TCDD)과 항생제의 일종인 Ketoconazole를 사용하였다. TCDD는 대부분의 식물체에 존재하는 flavonoid 물질과 마찬가지로 호르몬 교란물질의 일종으로써 estrogen receptor에 결합하며 cytochrome P450 side chain cleavage enzyme (P-450scc)에 의해서 prognenolone으로 轉換되는 過程을 막아서 testosterone의 合成, 혈액내 testosterone의 濃度, 睾丸 및 副睾丸의 發育 그리고 정자의 形成 등을 抑制한다. Ketoconazole은 항진균 물질로 사람과 동물에서 testosterone의 合成을 沮害한다. 枸杞雙和湯 煎湯液은 mount frequency, intromission frequency를 대조군에 비하여 유의하게 증가시켰으며, TCDD와 枸杞雙和湯 煎湯液을 함께 투여한 군에서 TCDD만을 투여한 군보다 mount frequency는 유의하게 증가시켰으며, ketoconazole과 TCDD에 의하여 저하된 혈청 testosterone 농도를 유의하게 증가시켰다. To evaluate effect of Gugissanghwatang water extract on the male sexual behavior and serum testosterone concentrations, experiments performed in male rats. The results were obtained as follows: 1. Mount frequency and intromission frequency were increased significantly after the administration of Gugissanghwatang water extract in normal rats. 2. Intromission frequency were increased significantly after the administration of Gugissanghwatang water extract in TCDD-treated rats. 3. Serum testosterone concentration were increased significantly after the administration of Gugissanghwatang water extract in ketoconazole-treated rats. 4. Serum testosterone concentration were increased significantly after the administration of Gugissanghwatang water extract in TCDD-treated rats.

AFB₁ 대사에서 phloretion의 이중 활성 효과

임대원,이광,고상상,진효연,은상용,최병민,김복량 圓光大學校 醫科學硏究所 2008 圓光醫科學 Vol.23 No.1

Aflatoxm B₁(AFB₁) is a potent hepatocarcinogen in experimental animals and a hazard to human health in several parts of the world. AFB₁ is activated to its ultimate carcinogenic intermediate, AFB₁-8,9-epoxide, by cytochrome P450(CYP) 1A2 and CYP3A4 in human liver and the intermediate is decomposed by several glutathione S-transferase(GST) including GSTA2, GSTM1 and GSTP1. In this study, we investigated the effects of phloretin on the enzyme systems which are involved in the activation and detoxification of AFB₁. The metabolic intermediate of AFB₁ was measured with HPLC. We found that phloretin could strongly inhibit the activities of CYP 3A4 and CYP1A2 in a dose dependent manner. Phloretin induced the antioxidant-response element(ARE)-mediated gene expression, including GSTs. The expressions of GSTA2, T1, M1, and GSTP1 were induced by 10μM phloretin. The decomposition of AFB₁-8,9-epoxide was measured with GSH conjugating activity of the epoxide. The rate was increased to 1.5 fold when HepG2 cells were treated by 10μM phloretin for 12h. In the mean while, the total GST activitives toward CDNB in HepG2 cells were not changed by the treatment with phloretin. The results demonstrate that phloretin has strong chemopreventive effects against AFB₁ toxicity through the inhibition of AFB₁ activation and induction of GSTs.

The Corrlation between Protein Methylation Ⅰ and Ⅲ Reaction and Immune Responses from Mouse Splenic Lymphocytes stimulated by Lipopolysaccharide

Kim, Bok Ryang 圓光大學校 醫科學硏究所 1989 圓光醫科學 Vol.5 No.1-2

LPS로 감각한 비장 임파구의 protein methylase Ⅰ 및 Ⅲ의 비활성은 각각 대조군의 3 및 4배로 증가하였으며, protein methylase Ⅰ은 LPS 처리후 36 시간에서, protein methylase Ⅲ는 72 시간에서 각각 최대의 활성을 보였다. 48 시간 동안의 비장 임파구 배양에서 protein methylase Ⅰ에 의한 arginyl 잔기의 methyl화 반응, protein methylase Ⅲ에 의한 lysyl 잔기의 methyl화 반응, DNA합성, 그리고 polyclonal 항체 형성 반응등이 SF보다 SIBA에 의해 더 저해되었다. SIBA에 의한 억제는 배양기간 중 언제라도 투여한 즉시에 나타났다. 반면 SF의 억제 효과는 배양 초기 12 시간 이내에 첨가해야 만이, 또 투여 후 36 시간이 지나야 저해효과가 보이기 시작했는데 그 후 급격히 증가하여 배양 72 시간 이후부터는 SIBA의 저해 효과보다 더 커져 protein methylase Ⅰ 및 Ⅲ의 in vitor 억제 정도와 일치하였다. 이들의 결과들은 protein methylase Ⅰ 및 Ⅲ가 LPS에 의한 비장 임파구의 면역기능 발현에 밀접하게 관여하고 있음을 시사하고 있다. The induction of protein methylase Ⅰ preceded protein methylase Ⅲ induction in the lipopolysaccharide-induced splenic lymphocyte cultures. The methylation of arginyl and lysyl residues, and the proliferative and polyclonal antibody responses at 48 hr of culture were inhibited more sensitively by SIBA. Regardless of when added, SIBA showed its effects on the responeses immediately after addition. The effects of SF were no longer observed when added later than the first 12 hr in the 48 hr experiment. At 96 hr of exposure, the effects were higher than those of SIBA. The data suggest that the methylation by these enzymes is associated with the immune responses of lymphocytes cultured with lipopolysaccharide.

Unscheduled DNA Synthesis Induced by Carcinogens in Primary Culture of Rat Hepatocytes

Kim, Bok-Ryang,Paik, Moon-Kee 圓光大學校 醫科學硏究所 1985 圓光醫科學 Vol.1 No.1

Rat의 1 차간세포 배양에 environmental 발암물질을 첨가하여 간의 DNA에 손상을 준뒤 복구되는 정도를 정량적으로 측정하였다. 즉, CaCl density gradient 을 초원심분리(ultra-centnfuge)로 만들어 DNA를 추출해서 그것에 결합된 [^3H]-thymidine양을 liquid scintillating 측정기로 측정하였다. Direct 발암물질인 MNNG에 의한 손상의 복구는 short patch 형태로 복구가 빨리 일어나고, indirect 발암물질인 Benzo(a) Pyrene에 손상의 복구는 large patch 형태로 복구가 천천히 일어난다. [^3H]-thymidine과 발암물질을 세포에 같이 넣고 UDS을 유도시키는 실험에서 B(a)P에 대한 손상의 복구는 12시간까지 계속해서 증가되었으나, MNNG에 대한 손상의 복구는 3시간안에 급속히 증가된 후 그 수준을 12시간 까지 유지시켰다. 발암물질을 세포에 넣어 2시간 동안 손상을 준후 [^3H]-thymidine을 세포에 첨가하여 UDS을 유도시키는 실험에서 B(a)P에 의한 복구는 실험 시작후 3-6 hr에서 최대를 보였다. 또 MNNG에 대한 실험에서는 0-3 hr에서 최대를 보였다. Indirect 발암물질은 세포내에서 대사가 되어야 DNA에 손상을 입힐수 있어 간세포를 얻은 후 18시간 이내에 실험을 시작해야 그 물질의 DNA와 결합하는 성질을 잘 나타낼수 있었다. Dose-response 실험에서는 MNNG는 10^-4M에서 B(a)p는 10^-5M에서 최대값을 얻었다. The ability of direct and pro-carcinogens to induce unscheduled DNA synthesis(UDS) in primary culture of hepatocytes was examined. Hepatocytes were isolated from adult male rats with collagenase perfusion technique and maintained in short-term monolayer culture on collagen-coated plates in serum free modified Waymouth's medium. Incorporation of [^3H]-thymidine into DNA in the presence of hydroxyurea was used to measure UDS. N-Methyl-N'-nitro-N-nitrosoguanidine(MNNG) which elicit a "short-patch" type of DNA repair and benzo(a) pyrene (B(a)P) which result in a "long-patch" type of DNA repair were used as standard carcinogens to show repair patterns. In continuous labelling experiments, UDS induced by B(a)P was increased linearly for 12hr in cultures, but that of

가물치(Channa asiatica)肝組織의 S-Adenosyl-L-Methionine : Protein Arginine N-Methyltransferase에 關한 硏究 Protein Arginine N - Methyltransferase of Snake Head(Channa asistica) Liver

崔宜奎,金福亮,白汶基 圓光大學校 醫科學硏究所 1986 圓光醫科學 Vol.2 No.2

Protein methylase Ⅰ(S-adenosyl-L-methionine : protein-arginine methyltransferase, EC 2.1.1.23) having an optimal pH at around 7.3 was Purified 117 fold with a 31% recovery from snake head liver. The enzyme showed a optimum temperature at around 3l℃ and was completely inactivated at 43℃. Both histone type Ⅱ-A and histone type Ⅷ-S were verified as good substrates in accepting methyl group from S-adenosyl-L-methionine , whereas histone type Ⅲ-s, egg albumin, r-globulin, bovine serum albumin, and lysozyme were poor. The Km value for S-adenosyl-L-methionine was 6.75×10 exp (-6) M at 37℃ and the maximum velocities were 47.6 and 84.8 pmole/min/㎎ protein at 21℃ and 31℃ respectively so that the activation energy of the enzyme was calculated as 21 .76 kJ/mole. Copper and zinc ions were the potent inhibitor, and the inhibitory effect were 43.5% and 33.8% of total activity at 200uM respectively. Sulfate ion also acted as an inhibitor, showing 50% of this enzyme activity at 160mM. Protein methylase I was not only activated but also stabilized the presence of ammonium ion.

白鼠 赤血球 膜 蛋白質의 카복실 메칠化 反應

嚴翼都,金福亮,白汶基 圓光大學校 醫科學硏究所 1986 圓光醫科學 Vol.2 No.2

The enzymatic carboxyl methyl esterification of red blood cell (RBC) membrane protein has been investigated in the isolated membrane and intact rat RBC population of different ages separated by density gradient centrifugation. The activity of cytosolic protein methylase Ⅱ was decreased according to the cell ageing, but isolated membrane protein from RBC of different age showed an age-related increased ability to act as methyl accepting substrates. Membrane proteins were shown to be the natural and excellent substrates for the cytosolic protein methylase Ⅱ compared with protein methylase Ⅰ and Ⅲ in the RBC cell so that the carboxyl methylation of membrane protein was the major methylating reactions in vitro and vivo. When RBC were incubated with media containing L-[methyl-^3H] methionine and 16.7 mM glucose, the [^3H]-methyl incorporation into the carboxyl group of membrane protein was 3.12 pmole/㎎ protein but incubated with media containing L-[methyl -^3H] methionine and 5.6 mM glucose, the amount of carboxyl methylation was 2.36 prnole/㎎ protein. About 1.4 fold increase in membrane protein carboxyl methylation was observed in intact younger RBC in comparison with intact older RBC in above media.