황기도홍탕의 抗腫瘍 效果 및 免疫調節反應에 관한 硏究

鄭遇悅,李鎭弘,田炳熏 대한동의병리학회 1996 동의생리병리학회지 Vol.10 No.1

황기도홍탕의 抗腫瘍 효과 및 면역조절작용을 입증하고자 MCA로 腫瘍을 유발하고 leukemia cell line인 3LL 세포와 sarcoma cell line인 S-180 세포 및 FsaⅡ 세포를 마우스의 背部 皮下에 이식함으로써 발생되는 腫瘍의 發生率 및 크기등을 평가하고, 동시에 생체내 免疫過敏性 반응인 Arthus 반응 및 DTH반응, 그리고 抗體 生産能, 시험관내 NK세포의 활성도, 정상림프구와 mitogen으로 자극한 癌腫細胞의 增殖 반응, 림프구의 lymophokine 生産能등을 관찰하여 다음과 같은 결과를 얻었다. 황기도홍탕 추출액은 MCA로 유도한 腫瘍의 발생률과 크기를 감소시켰고, 3LL 세포 및 s-180세포 이식에 의한 腫瘍 발생을 감소시켰으나, Fsall 세포 이식에 의한 腫瘍발생에서는 영향을 미치지 못하였다. 또한 황기도홍탕 추출액은 시험관내 A431 세포의 증식에는 영향을 미치지 못하였으나, Fsall 세포의 增殖든 도리어 항진시켰고, 3LL 세포 및 S180 세포의 增殖은 저하시켰다. 황기도홍탕 추출액은 NK세포의 활성도를 증가시켰으며, 면역적혈구에 대한 마우스의 Arthus 반응 및 DTH 반응, 그리고 赤血球 凝集素의 免疫 반응들을 모두 항진시켰다. 황기도홍탕 추출액은 mitogen자극 램프구의 增殖能을 항진시켰으며 그 항진은 mitogen으로 세포를 자극함과 동시에 황기도홍탕 추출액에 노출시켰을 때 더욱 항진되었다. 황기도홍탕 추출액은 림프구의 IL-2 및 MIF 生産能에서는 영향을 미치지 못하였다. 이상의 결과로 보면 황기도홍탕의 종양효과와 면역조절반응은 MCA와 3LL 세포 및 S-180 세포로 유도된 皮下癌腫 세포에 대하여 특이적 細胞 毒性과 免疫調節作用에 의한 것으로 볼 수 있다. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Hoangkidohongtang extract against tumor. and to carry out immunomodulatory effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene(MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S-180 and Fasll cells). Treatment of the Hoangkidohongtang extract (daily 1 mg/mouse, i. p.) was continued for 7 days prior tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Hoangkidohongtang decreased not only the frequency of tumor production but also the numbers and the weight of tumors per tumor bearing mice (TBM). Hoangkidohongtang also significantly suppressed the development of 3LL cell and S-180 cell by frequency and their size, and some developed tumors were regressed by the continuous treatment of Hoangkidohongtang extract into TBM. However, when tumor was induced by Fsall cell implantation, the growth of implanted cells in mice was delayed by the water extract of Hoangkidohongtang until day 7 and then rapid growth ensued. in vitro, treatment of Hoangkidohongtang extract had no effect on the growth of some kind of cell lines such as Fsall, A431 strain but significantly inhibited the proliferation of 3LL, S-180 cells. Hoangkidohongtang also stimulated the migrative ability of leucocyte, the MIF and IL2-production of T lymphocytes, but not IL6 production of B cells. Hoangkidohongtang enhanced Arthus reaction and DTH to sheep erythrocytes, and NK cells activities. These results demonstrated that Hoangkidohongtang extract exhibited a significant prophylatic benefits against tumors and showed different results according to the type of tumor cells. And these results also suggested that antitumor effect of Hoangkidohongtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

궁귀탁리산(芎歸托裏散)의 L1210과 S-180이 이식된 마우스에 대한 항암(抗癌) 작용(作用) 연구(硏究)

박수연,김종한,최정화,박용호,Park, Su-Yeon,Kim, Jong-Han,Choi, Jung-Hwa,Park, Yong-Ho 대한한방안이비인후피부과학회 2006 한방안이비인후피부과학회지 Vol.19 No.1

Objective : The purpose of this study was to investigate effect of Gungguitakli-San(GTS) on the anti-tumor, immunocytes. Methods : This study estimated the proliferation of L1210 and S-180 cell lines, mouse splenocytes and thymocytes in vitro, and estimated the proliferation of L1210 cell, S-180 cell, thymocytes and splenocytes and body weight in S-180 cells-transplanted mice. The cytotoxicity and proliferation of cells were tested using a colorimetric tetrazoliun assay(M1T assay). Results : The results of this study were obtained as follow ; 1. GTS was significantly increased in the proliferation of thymocytes and splenocytes In vitro. 2. GTS was significantly showed cytotoxicity on the L1210 cell lines and 8-180 cell lines in vitro. 3. GTS was significantly showed cytotoxicity on the L1210 cell lines in vivo. 4. GTS was significantly increased in the weight of mice and decreased weight of sarcoma, in S-180 cells transplanted mice. 5. GTS was significantly increased in the period of survive, in S-180 cells transplanted mice. Conclusions : The author thought that GTS had action of anti-cancer by becoming immunocytes activity and by cytotoxicity of cancer cells.

膈下逐瘀湯이 癌細胞의 增殖에 미치는 實驗的 效果

權晟五,鄭鉉雨 대한동의병리학회 2000 동의생리병리학회지 Vol.14 No.1

膈下逐瘀湯은 『醫林改錯』에 "治瘀在膈下, 形成積塊..."라고 최초로 수록된 방제로 活血化瘀의 代表方劑로 인식되면서 下焦氣瘀停患과 瘀血內結로 인한 積0등에 이용되어 왔다. 癌은 지금까지 알려져 있는 사망원인중 높은 비율을 차지하고 있는 질환으로 東醫學에서는 飮食下節, 七情0結 및 臟腑의 機能失調 등으로 인하여 發顯된 體內腫塊를 指稱하였고, 그 發生棋戰, 대해서는 氣血虧虛·氣滯血瘀·痰結00·熱0內蘊·經結瘀阻 등에 의한다하였다. 또한 西양醫學에서는 조직의 자율적인 과잉 성장으로 개체에 대해 이롭지 않을 뿐만 아니라 정상 조직에 대해서도 파괴적인 세포조직으로 비정상적인 세포증식을 말하였다. 현재 抗癌에 관한 연구와 膈下逐瘀湯을 이용한 연구가 활발히 진행되고 있지만 膈下逐瘀湯에 대한 연구는 주로 瘀血의 病態모텔에 대한 報告로 抗癌0果는 미미한 실정이다. 이에 著者는 다양한 種瘍의 治療法중 韓方의 活血化瘀法을 이용하여 그 효과를 확인하고자 膈下逐瘀湯을 선택한 후 L1210세포와 S-180세포를 이용하여 L1210세포의 증식능, L1210세포가 이식된 동물의 복강 macrophaes에서 생성되는 NO의 量 그리고 S-180세포로 유발된 동물의 고형암 무게, 체중, 생존기간 연장 및 혈액내 RBC, WBC, HGB, HCT 등을 측정하였던 결과 膈下逐瘀湯이 복강 macrophages에서 생성되는 NO와 밀접한 관련을 갖고 있음으로서 L1210세포에 대한 직·간접적인 抗癌0果를 나타내었고, 또한 固形癌과 생명연장 효과들을 관찰한 결과에서도 우수한 효과가 있는 것으로 나타났다. 그러나 생체내 변화중 혈액내의 변화는 앞으로 더욱 연구를 해야할 것이다. Keukhachookeotang(KCT) has been used in Oriental Medicine as drug for promoting blood circulation and removing blood stasis. The purpose of this Study was to investigate effects of KCT on the proliferation of L1210 cells, Nitric Oxide(NO) production from peritoneal macrophages in L1210 cells-transplanted mice, and on the tumor weight, body weight, survival time and blood changes in S-180 cells-transplanted mice. We were used KCT extract with freeze-dried, 8wks-old male mice, L1210 cell lines, S-180 cell linse for this Study. The cytotoxicity of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follow;KCT ingibited proliferaion of L1210 cells and accelated NO production from peritioneal macrophages in L1210 cells-transplanted mice. KCT was decreased significantly tumor weight and body weight, and increased survival time in S-180 cells-transplanted mice. In blood changes KCT decreased significanfly on the RBC, WBC, HGB and HCT in S-180 cells-transplanted mice.

Acetylshikonine 및 합성 Naphthazarin 유도체의 L1210 및 S-180 암에 대한 항암효과

김현,안병준 忠南大學校 癌共同硏究所 1991 癌共同硏究所 硏究誌 Vol.1 No.1

Acetylshikonine, isolated from the root of Lithospermum erythrorhizon showed a strong cytotoxic activity (ED_(50)=0.10 ug/m/) against L1210 cell and T/C=182% in ICR mice bearing S-180 at a dose of 5 mg/kg. Administrations of 10 mg/kg and 15 mg/kg reduced the T/C values to 60 and 77% respectively. Higher doses reveal toxicity. Seven naphthazarin derivatives synthesized showed good cytotoxic activities against L1210 cell. Especially, naphthazarin and hydronaphthazarin have strong activities (ED_(50)=0.05 ug/m/ for both). Naphthazarin showed a severe toxic effect on ICR mice bearing S-180; no significant toxic effect was observed at a dose of 1 mg/kg or 2 mg/kg, but a severe toxicity (T/C=23%) by administration of 5 mg/kg. Alkylation of C-2 of naphthazarin is necessary for reducing the toxic effect on ICR mice bearing S-180.

Acetylshikonine 및 합성 Naphthazarin 유도체의 L1210 및 S-180암에 대한 항암효과

김현,안병준 忠南大學校 癌共同硏究所 1991 癌共同硏究所 硏究誌 Vol.1991 No.-

지모(知母) 추출물이 L-1210 및 S-180 암세포주 성장 억제에 미치는 영향

임치혜,초재승,김효수,권승만,김신,김일환,박혜선,Yim, Chi-Hye,Cho, Jae-Seung,Kim, Hyo-Soo,Kwon, Seung-Man,Kim, Shin,Kim, Il-Hwan,Park, Hye-Sun 사상체질의학회 2007 사상체질의학회지 Vol.19 No.2

1. Objective This study was aimed to screen the potential antitumor activity of one kinds of Korean medicinal herb extracts against cancer cell lines and to evaluate the growth inhibition effect of L-1210 and S-180 cancer cell lines. 2. Methods It confirmed Anemarrhena asphodeloides extracts to screen the potential antitumor activity. Then, it was extracted with 4 kinds of solvents ; hexane, ethyl acetate, butanol and $H_2O$, and the Growth inhibition effect of these extracts were determined against cancer cell and normal cell. The results were as follows : The IC50(50% inhibitory concentration) values of Anemarrhena asphodeloides extracts were shown to be $253{\mu}g/ml$ against L-1210 cell lines. The IC50 values of ethyl acetate extracts were shown to be $915{\mu}g/ml$ against L-1210 cell lines. The IC50 values of butanol extracts were shown to be $52.3{\mu}g/ml$, $485{\mu}g/ml$ against L-1210, S-180 cell lines, respectively. The butanol extracts were more selectively effective than other extracts to cancer cell lines. 3. Conclusion From these data, it could be concluded that the Anemarrhena asphodeloides extracts to the Growth inhibition effect of L-1210 and S-180 cancer cell lines.

Acetylshikonine 및 합성 Naphthazarin 유도체의 L1210 및 S-180 암에 대한 항암효과

김현(Hyun Kim),안병준(Byung Zun Ahn) 대한약학회 1990 약학회지 Vol.34 No.4

Acetylshikonine, isolated from the root of Lithospermitm erythrorhizon showed a strong cytotoxic activity (ED50=0.10mcg/ml against L1210 cell and T/C = 182% in ICR mice bearing S-180 at a dose of 5mg/kg. Administrations of 10mg/kg and 15mg/kg reduced the T/C values to 60 and 77% respectively. Higher doses reveal toxicity. Seven naphthazarin derivatives synthesized showed good cytotoxic activities against L1210 cell. Especially, naphthazarin and hydronaphthazarin have strong activities (ED50=0.05mcg/ml for both). Naphthazarin showed a severe toxic effect on ICR mice bearing S-180: no significant toxic effect was observed at a dose of 1mg/kg or 2mg/kg, but a severe toxicity (T/C = 23%) by administration of 5mg/kg. Alkylation of C-2 of naphthazarin is necessary for reducing the toxic effect on ICR mice bearing S-180.

桔梗湯과 加味桔梗湯의 抗癌效果와 免疫反應에 關한 硏究

宋孝貞,金聖勳,李光浩 대한동의병리학회 1996 동의생리병리학회지 Vol.10 No.1

肺癰에 使用되고 있는 桔梗湯을, 肺癌株 및 S-180에 投與하여 抗癌 抗轉移 및 免疫效果를 實驗的으로 糾明하고, 여기에 現在 肺癌에 多用되고 있는 2가지 藥物을 加味하여 相乘效果를 觀察하고자 하였다. C57BL/6에 B16-Fo를 皮下主射하고 ICR생쥐에 S-180을 腹腔 注射한 後 桔梗湯과 加味桔梗湯을 投與하여 腫瘍重量과 體重變化, 生命延長率, 組織變化 및 血液學的變化 등을 測定하여 抗癌效果를 分析하고, 아울러 FACS에 의해 BALB/c의 脾臟免疫細胞에 대한 影響을 살펴보았다. 桔梗湯, 桔梗湯加腥草 및 桔梗湯加白花蛇舌草등은 모두 抗癌, 抗轉移 및 免疫反應에서 有意性있는 結果를 認定되었으며, 魚腥草나 白花蛇舌草를 加味하였을 경우 더욱 效果的으로 나타났다. In order to prove the antitumor effect of Kilkyungtang(KKT), Kilkyungtang with Houttuyniae Herba(KKT-I) and Kilkyungtang with Oldenlandiae diffusae Herba(KKT-Ⅱ) experimentally, the studies were done. We measured tumor mass and body weight hematological change in ICR bearing S-180 and C57BL/6 implanted with B16-Fo, also studied their effect on immune cells in BALB/c by FACS. The results were obtained as follows: 1. Body and tumor weight were significantly suppressed in all groups as compared with control group. MST(mean survival time) was 11% in KKT, 16.9% in KKT-Ⅰ, 16.1% in KKT-Ⅱ for 14 days after implantation s.c. with B16-Fo and also body weight was significantly suppressed after implantation i.p. with S-180, MST was 20.1% in KKT, 40.9% in KKT-1, 56.6% KKT-Ⅱ for 62 days. 2. WBC and platelet aggregation was significantly increased in all groups, the number of platelet was significantly increased in KKT and KKT-Ⅰ as compared with control group. 3. The infiltration of tumor cell was obvious in the white and red pulps, in the sinusoid of liver and the glomeluli of kidneys after implantation with S-180, whereas the changes of tissues were not significant in KKT-Ⅱ, KKT-Ⅰ and KKT in order. 4. T cell and T helper cell were increased in KKT-Ⅰ and KKT-Ⅱ treated groups and B cell was increased in KKT treated group as compared with control group. From above results it was concluded that Kilkyungtang(KKT), kilkyungtang with Houttuyniae Herba(KKT-Ⅰ) and Kilkyungtang with Oldenlandiae diffusae Herba(KKT- Ⅱ) had antitumor effect, and also Kilkyungtang combined with Houttuyniae Herba or Odenlandiae diffusae Herba were more effective than KKT only which suggests the necessity of continuous study on the effects of Houttuyniae Herba or Oldenlandiae diffusae Herba.

Effect of Low Dose Mutagens on Adoptive Response and Plasma Membrane Glycoconjugates in Sarcoma 180 Cells

Lee, Jong Hwa,Choi, Eun Sili,Oh, Kyu Seon,Lee, Dong Wook,Chang, Jeong Hyun,Um, Kyung Il The Korean Society for Integrative Biology 2000 Korean journal of biological sciences Vol.4 No.3

The present investigation was performed to elucidate the effect of pretreatment with low dose ultraviolet radiation (UV) and ethyl methanesulfonate (EMS) on cell survival by trypan blue dye exclusion method and plasma membrane glycoconjugates by lectin-cytochemistry in sarcoma 180 (S180) cells. Pretreatment with 2 J/$m^2$ UV or 2 mM EMS increased the percentage of survival of cells subsequently treated with high dose UV (10 or 20 J/$m^2$) or EMS (10 or 20 mM), respectively. Staining intensity of concanavalin A (Con A) of the cells pretreated with 2J/$m^2$ UV or 2 mM EMS and subsequently treated with 10 or 20 mM EMS was stronger than that of the cellstreated with 10 or 20 mM EMS. These results suggest that there is an adaptive response on cell survival to EMS or UV in S180 cells. And the results show a change in mannose-containing glycoconjugates of plasma membrane in S180 cells pretreated with EMS or UV and subsequently treated with EMS.

신령버섯균사체 액체배양물의 자가분해에 의한 항암성 isoflavone-conjugated glycoprotein 분리

김소영(So Young Kim),김영숙(Young Suk Kim),장정순(Joung Soon Jang),김보현(Boh Hyun Kim),라키브 압두르(Abdur Md. Rakib),김곤섭(Gon Sup Kim),김정옥(Jeong Ok Kim),하영래(Yeong Lae Ha) 한국생명과학회 2014 생명과학회지 Vol.24 No.12

신령버섯균사체(Agaricus blazei mycelia: ABM)를 대두박이 함유된 액체배지에 배양하고, 이것을 자가분해(53 ℃, pH 5.5, 120 rpm, 3 hr)하여 항암성이 강한 isoflavone-conjugated glycoprotein (Gluvone 이라 명명)을 분리하였다. Gluvone 은 지금까지 알려진 당단백질과는 달리 분자량이 작고(9,400 Da), isoflavone이 결합되어 있다는 점이 다르다, Gluvone 은 60% 탄수화물(glucose, fructose, ribose), 31% 단백질 및 2% isoflavone (daidzein, genistein)으로 구성되어 있었다. 이 Gluvone은 S-180 복수암세포, MCF-7 인체유선암세포에 대한 독성이 강하였고, S-180 세포로 유발한 mouse 복수암을 강하게 억제하였다. Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone- conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at 53℃ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with ¹H-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.