사군자탕가산자고가 위암세포에 미치는 항암효과에 대한 분자생물학적 연구

류봉하,류기원,윤상협,김진성,김진석,Ryu, Bong-Ha,Ryu, Ki-Won,Yoon, Sang-Hyub,Kim, Jin-Seong,Kim, Jin-Seok 대한한방내과학회 2002 大韓韓方內科學會誌 Vol.23 No.2

1. Background The previous studies on anticancer medicine derived from korean traditional medicine have focused on the life elongation of cancer cell bearing animals. However, it is thought that more molecular biological studies are needed to reveal their mechanism. 2. Objective The aim of this study was to investigate the molecular biological function of Sagunjatang plus Cremastrae Appenediculatae Tuber on cytostaticity, apoptosis and apoptosis related genes revelation against human stomach cancer cells(AGS). 3. Methods After administrating Sagunjatang and Sagunjatang plus Cremastrae Appenediculatae Tuber to human stomach cancer cell. MTT assay was performed to compare and examine the efficacy of each medicine on the cytostaticity of stomach cancer cells in proportion to time and doses, and apoptosis assay was performed to examine their effect on apoptosis by using DAPI dye and counting the number of cells which developed in an apoptotic body. In addition, the quantitative RT-PCR was used to examine their effect on the revelation of Bcl-2, Bax and P53, which are genes related to apoptosis. 4. Result and Conclusion Sagunjatang plus Cremastrae Appenediculatae Tuber demonstrated increased cytostaticity. decreased apoptosis and unremarkable revelation of apoptosis related genes. But in the cytostaticity and apoptosis, Sagunjatang plus Cremastrae Appenediculatae Tuber showed a tendency to control stomach cancer cells. Therefore, we can expect the clinical application to the related diseases. Besides, it needs another experiment on various cancer cells, such as, lung cancer cell and hysterocarcinoma cell.

사매가 위암세포에 미치는 영향

류봉하,김진성,윤상협,류기원,홍상선,Ryu Bong-Ha,Kim Jin-Sung,Yoon Sang-Hyub,Ryu Ki-Won,Hong Sang-Sun 대한한방내과학회 2003 大韓韓方內科學會誌 Vol.24 No.2

Background : Nowadays many researches about it s cure are going on world widely since cancer is one of the most human health threatening diseases. In Chinese and North Korean medicine, Duchesnea india(Audra.) Foche. is practically used to treat many kinds of cancer, but in Korea it is rarely used. So, we need to scientifically identify anti-tumor effects of Duchesnea india(Audra.) Foche. Objective : We are aimed to identify anti-tumor effects of Duchesnea india(Audra.) Foche. on the stomach cancer cells through molecular biologic methods. Material & Methods : We used AGS as stomach cancer cells from American Type Culture Collection. We added the boiled extract of Duchesnea india(Audra.) Foche. $5{\mu}l$(Sample I), $10{\mu}l$(Sample II) to cultural media(ml)for 0,6, 12, 24, 48 hours. We measured the killing effect on stomach cancer cells through Tryphan blue exclusion test and the suppressive effect on viability of stomach cancer cells via MTT assay. the quantitative RT-PCR was used to examine their effect on the revelation of Bcl-2, Bcl-XL, and Bax, which are genes related to apoptosis. We measured change of mitochondria membrane permeability and membrane potential via flow cytometry. Result : 1. The killing effect on stomach cancer cells showed that each test groups killed more stomach cancer cells than the control group with a time(6 hours later) and density dependent manner, which was statistical significance. 2. The suppressive effect on viability of stomach cancer cells showed that each test groups had more suppressive effects on viability of stomach cancer cells than the control group with a time(6 hours later), which was statistical significance. 3. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 and Bcl-XL decreased with a density manner which was statistical significance. but the revelation of Bax was not changed with statistical significance. 4. As a result of this test, Duchesnea india(Audra.) Foche. caused apoptosis by decreasing the absorbance of mitochondria with statistical significance. and also induced apoptosis by decreasing the membrane potential of mitochondria. Conclusion : This experiment showed that Duchesnea india(Audra.) Foche. has anti-tumor effect with statistical significance. This is in vitro experiment and basic experiment on Duchesnea india(Audra.) Foche. We hope more progressive researchs on Duchesnea india(Audra.) Foche. will go on and its anti-tumor effects will be more practically identified.

변비(便秘)에 관(關)한 동서의학적(東西醫學的) 고찰(考察)

류봉하,조남희,Ryu, Bong-Ha,Cho, Nam-Hee 대한한방내과학회 2000 大韓韓方內科學會誌 Vol.21 No.1

Objectives : To satisfy the demand of good treatment of constipation Methods : we investigated the literatures of Oriental Medicine about Constipation. Results: 1. There are three categories of etimological factors of constipation, that is, endogenous, exogenous and non-exo-endogenous factor. The endogenous factor is caused by seven emotions, called depression of Ki and stagnation of Ki. The exogenous factor is six excessive atmospheric influences, for example, wind, cold, dampness, heat and dryness. And the non-exo-endogenous factors are overfatigue, improper diet, stagnated blood and deficiency of Ki and blood that comes from old age, long disease and after delivery. 2. Classification: According to cause of disease it is classified by constipation due to cold, heat, wind, dryness, retention of undigested and phlegm. According to Internal Organs there are constipation due to deficiency syndrome of the stomach, excess syndrome of the stomach, deficiency syndrome of kidney and splenic constipation. And Differentiation of syndromes according to Ki and blood, there are constipation of deficiency type and excess type. There are constipation due to stagnation and deficiency of Ki, deficiency of blood and stagnated blood. 3. Principles and Methods of treatment 1) Herbal Medicine (1) Excess type [1] Constipation due to heat : Seunggitang(承氣湯) and Majainwhan(麻子仁丸) [2] Constipation due to stagnation of Ki : Samatang(四磨湯) and Yukmatang(六磨湯) (2) Deficiency type [1] Constipation due to deficiency of Ki : Whanggitang(黃?湯) [2] Constipation due to deficiency of blood: Yunjangwhan(潤腸丸) [3] Constipation due to cold : Jechunjun(濟川煎) and Banyuwhan(半硫丸) 2) Enema therapy: It is a method to induce defecation by honey or pig's bile juice for weak people. 3) Acupuncture and Moxibustion: Acupoints used to treat constipation are BL25, ST25 and TE6. Moxibustion at CV8, CV6 is good for constipation due to cold. (4) Diet therapy: It is very important that we eat meals regularly and defecate on the same time even if you don't. And we have to eat food like fruits, vegetables and beans. (5) finger pressure: Finger pressing around these points like ST25, SP25, BL25, BL31, BL32, BL33 and BL34 is good for it. (6) Kigong therapy: Abdominal breathing (7) Old man' s constipation: Hip bath or diet therapy is commended. Laxation with lubricant like Supungyunjangwhan(搜風潤腸丸) is used. (8) Women' s constipation: After delivery, we have to administer tonifying prescription Sipjundaebotang(十全大補湯) and enema can be used. Conclusion : We have to examine the cause of disease and bowl movement carefully. After comprehensive analysis of the data gained by the four methods of diagnosis, we diagnose and treat patients on the base of overall of symptoms and signs.

황약자(黃藥子)의 위암세포(胃癌細胞)에 대(對)한 항암(抗癌) 효능(效能) 검색(檢索)

류봉하,류기원,윤상협,김진성,김성희,Ryu, Bong-Ha,Ryu, Ki-Won,Yoon, Sang-Hyub,Kim, Jin-Seong,Kim, Sung-Hee 대한한방내과학회 2002 大韓韓方內科學會誌 Vol.23 No.1

Background : Nowadays a lot of research is based on natural substances or materials world wide since many kinds of side effects are accompanied by anti tumor chemotherapy. In Chinese medicine, Dioscorea bulbifera L is widely used to treat many kinds of cancer, but in Korea it is rarely used. Therefore, we need to scientifically identify anti tumor effects of Dioscorea bulbifera L. Objective : We aimed to identify anti tumor effects of Dioscorea bulbifera L on the stomach cancer cells through molecular biological methods. Materials & Methods : We used AGS, a stomach cancer cell from American Type Culture Collection. We injected the boiled extract of Dioscorea bulbifera L 5 ul(sample 1), 10ul(sample 2) to cultured media(ml) for 0,6,12,18,24 hours. We measured the killing effect on stomach cancer cells through Tryphan blue exclusion test and suppressive effect on viability of stomach cancer cells via MTT assay. Results : Tryphan blue exclusion test showed that each test group killed more stomach cancer cells than the controlled group with a dosage-dependent, but not significantly. MTT assay showed that each test group had a more suppressive effect on viability of stomach cancer cells than the controlled group without a dosage-independent, but not significantly. The cell cycle analysis via flow cytometry showed that the test group extended cell cycle, and there was no peak in M phase, the number of sub G1, G0, G1 phase cells increased a little, but not significantly. Conclusion : This experiment showed that Dioscorea bulbifera L. has an anti-tumor effect, but not significantly. This is in vitro experiment and basic experiment on Dioscorea bulbifera L. We hope more progressive researches on Dioscorea bulbifera L. will be conducted and its anti tumor will be more accurately identified.

단삼보혈탕(丹蔘補血湯)이 Stress로 유발(誘發)된 백단삼보혈탕(白丹蔘補血湯)이 Stress로 유발(誘發)된 백서(白鼠) 위장관(胃腸管)의 조직손상(組織損傷)에 미치는 영향(影響)

류봉하,박동원,류기원,공현식,Ryu, Bong-Ha,Park, Dong-Won,Ryu, Ki-Won,Kong, Hyun-Sig 대한한방내과학회 2000 大韓韓方內科學會誌 Vol.21 No.1

Objectives : This is the experimental paper to study the curative and preventive effects of Dansambohyultang, Methods : Oriental herb medicine for gastro-intestinal disease caused by stress, on the rats put under the stress through starvation, heating and immoblization, the author used four different group; the normal group, which was not put under stress and well fed, and the control group, to which the drug was not administered and put under the stress through starvation, heating and immoblization, the Sample I, to which the drug was administered before they were put under stress, the Sample lI, to which the drug was administered after they were put under stress. Results : When a Masson's trichrome stain method was applied to the control group, a small size of ulcer was found in the fundus and the pylorus, and atrophy was observed in the neck region of mucous membrane. When the drug was administered to the Sample I and the Sample II, the former recovered from the ulcer and atrophy to almost normal and showed better results than the latter. When a Masson's trichrome stain method was applied to the control group, atrophy of mucous membrane was found all over the intestines. When the drug was administered to the Sample I and the Sample ll, the curative and recovered effects was seen in both, but the former showed better results than the latter. After application of Mayer's hematoxylin stain method, the observation of numerical changes of goblet cells in the small intestines showed the most significant decrease in the control group. In the Sample I, the number of the cells in the duodenum, the jejunum and the ileum returned to an almost normal level. In the Sample II, a slight increase was observed only in the ileum. After application of Mayer's hematoxylin stain method, the observation of numerical changes of goblet cell in the large intestines showed a significant decreased in the control group. Both the Sample I and the Sample II showed an increase in the number of goblet cells in comparison with the control group. When a serotonin-immunoreaction test was applied, the control group showed a general decrease in the number of gastro-entero-endocrine cells, The Sample I showed the effect in all over gastro.intestinal tract as compared with control group and the Sample II showed the effect in all over gastro-intestinal tract except the duodenum and the former showed better results than the latter. Conclusion : According to the above results, the regions which are most sensitive to stress were observed to the ileum and the colon. It was concluded that Dansambohyultang has a remedical value particularly on the abnormalities caused by stress in all over gastro-intestinal tract and it was found to be more effective in the Sample I than in the Sample ll. Judging from the fact that the Sample I showed better results than the Sample ll, Dansambohyultang can be said to have a preventive effect more than a curative effect on gastro-intestinal tract disease.

보심건비탕(補心健脾湯)과 정전가미이진탕(正傳加味二陳湯)의 적출장기(摘出臟器), 항궤양(抗潰瘍), 위액(胃液)·gastrin 분비(分泌), 장관수송능(腸管輸送能) 및 진통(鎭痛).진경작용(鎭痙作用)에 대한 비교(比較) 연구(硏究)

류봉하,박동원,류기원,윤상건,김진성,Ryu, Bong-Ha,Park, Dong-Won,Ryu, Ki-Won,Yin, Chang-Jina,Kim, Jin-Seong 대한한방내과학회 2000 大韓韓方內科學會誌 Vol.21 No.1

Objectives : This is the experimetal paper to evaluate and compare the effects of Bosimgunbitang(補心健脾湯) with those of Jungjungamiyijintang(正傳加味二陳湯) on contraction of isolated organs, anti-ulcer, secretion of gastric juice, secretion of gastrin in serum, transport ability of intestine, analgesic effect and sedative effect Methods : We used mice and rats administered with the extract of the above herbs. Results: Bosimgunbitang and jungjungamiijintang showed the inhibitory effect on the smooth muscle contraction of the isolated ileum induced by acethylcholine chloride and barium chloride in mice. transport ability of intestine. Bosimgunbitang and jungjungamiijintang showed the inhibitory effect on the contraction of the fundus-strip induced by acethylcholine chloride and barium chloride in rats. The preventive effect on pyloric ulcer, indomethacin induced ulcer and ethanol-HCl induced ulcer bf mice was significant. The inhibitory effect of Bosimgunbitang on gastric free acidity, total acidity in Shay rats was significant. The inhibitory effect of on gastrin secretion in serum was significant after only 1 hour when the Bosimgunbitang and jungjungamiijintang was administered. The promoting effect of the jungjungamiijintang on the transport ability of small intestine was significant in the high concentration. The promoting effect of the Bosimgunbitang and jungjungamiijintang on the transport ability of large intestine was significant. The analgesic and the sedative effect were recognized. It meaned that Bosimgunbitang and jungjungamiijintang effected on the C.N.S.. Conclusion : Bosimgunbitang and jungjungamiijintang have the inhibitory effect on the smooth muscle abnormal contraction of the isolated ileum and the anti-ulcer effect to prevent, secretion of gastric juice and gastrin in serum. And they also have to increase a transport ability of intestine, analgesic effect and sedative effect. Jungjungamiijintang especially is eminent for analgesic effect while Bosimgunbitang has excellent sedative effect.

기능성 소화불량증 환자에 대한 반하사심탕 엑기스제의 임상효과

류봉하,류기원,김진성,윤상협,Ryu Bong-Ha,Ryu Ki-Won,Kim Jin-Sung,Yoon Sang-Hyub 대한한방내과학회 2003 大韓韓方內科學會誌 Vol.24 No.2

Objective : We evaluated the therapeutic effect of Banwhasasimtang dry extract powder on gastrointestinal symptoms and its side effects in patients with functional dyspepsia. Methods : Patients with functional dyspepsia, defined as discomfort in the epigastrium. A total 45 patients participated in this study. Dyspeptic symptoms ratio was obtained by patients, total point of symptom score/total 21 points x 100(%). On each of symptom after 4 weeks administration of Banwhasashim-tang was made by variation of symptom score: If symptoms score of posttreatment is lower than that of pretreatment, it is improved ; Otherwise aggravated, and if there is no change between them, unchangeable. When a full health condition of normal control is assumed 100%, relative health condition of dyspeptic patient could be calculated from 'full health condition-dyspeptic symptoms ratio'. Therefore a whole satisfaction rate on symptom improvement was presented with variation of a relative health condition to dyspepsia between pretreatment and post. Results : Banwhasashimtang significantly improved symptoms in patients. The significance was shown in both all seven items(p<0.01, respectively) and 19.2% (p<0.01) of increase of a whole satisfaction rate on symptom improvement. There is no remarkable side-effect in all laboratory test, excepting 1 case of abdominal pain and another one case of recurred pain of chronic prostatitis. Conclusion : Banwhasashimtang extract provides symptomatic relief in functional dyspepsia and is no remarkable side-effect drug.

소암거담익기탕(消癌去痰益氣湯)의 cyclophosphamide 부작용 형성 억제효과

류기원,류봉하,윤상협,김진성,홍종희,Ryu, Ki-Won,Ryu, Bong-Ha,Yoon, Sang-Hyub,Kim, Jin-Seong,Hong, Jong-Hee 대한한방내과학회 2002 大韓韓方內科學會誌 Vol.23 No.1

Objective : It is well known that modern chemotherapy against cancer has side effects to a living body, especially hemopoietic and immunologial disfunctions. However, there are no effective ways to reduce them. Recently, traditional Korean herb medicine has been reported to have some biological modifying responses. Therefore, we hypothesized that additional application of herb medicine during chemotherapy is more effective to reduce its side effects. While we were studying the effects, we have observed the inhibitory effect of Soamgudamikgitang on formation of side effects derived from Cyclophosphamide, it has been used in clinical practice at Kyung Hee Medical Center. Methods : We injected 200mg/kg of Cyclophosphamide, one time, to an experimental group, consisting of ten mice. We divided them into eight groups: normal, CPX, SAKT 2mg, SAKT 10mg, SAKT 50mg, SAKT 2mg, CPX, SAKT 10mg+CPX, SAKT 50mg+CPX. We injected Soamgudamikgitang seven days, five days, three days, and one day before we injected CPX. One day, three days, and five days after CPX injection, we injected Soamgudamikgitang again and then killed all the mice. The parameters determined in this experiment were daily body weight liver and spleen weight, RBC, WBC, and platelet for hemopoietic dysfunction and AST, ALT for hepatotoxicity, BUN, creatine for renal toxcity, lymphocyte proliferation activity and lymphocyte subsets for immunological toxcity. Results : We have found that Soamgudamikgitang has inhibitory effects on the formation of Cyclophosphamide's side effects. Significant differences between the group, which contained Cyclophosphamide, and the other group, which contains Cyclophosphamide and 2, 10, 50mg of Soamgudamikgitang respectively were observed. Platelets(2mg of Soamgudamikgitang, p<0.05 ;10mg, p<0.01 ;50mg, p<0.001), liver weight(50mg, p<0.01), spleen weight(10mg, p<0.05), AST(all groups, p<0.01), ALT(2mg, p<0.01 ;10mg, p<0.05 ;50mg, p<0.01), BUN(2mg, p<0.01 ;50mg, p<0.05). Although immunological in both lymphocyte proliferation and its subsets were not observed, which shows that Soamgudamikgitang has a strong effect on T cell activities. Conclusions : From the above results, we can expect that the combined therapy of Soamgudamikgitang and Cyclophosphamide is more effective for treating cancer patients.

藿香正氣散과 少陰人藿香正氣散의 效能에 關한 實驗的 比較硏究

柳基遠,柳逢夏,尹喜植,朴東源 慶熙大學校韓醫科大學韓醫學硏究所 1998 慶熙韓醫大論文集 Vol.21 No.1

To evaluate and compare the effects of Kwakhyangjeonggisan and Souminkwakhyangjeonggisan, the effect on the extracted organ, on the ulceration, on the gastric secretion, on the intestinal movement, on the diarrehea and on the temperature were studied using mouse administered with herbal extract. The results are as follows. 1. In the effect on the extracted organs, both two prescriptions showed concentration-dependent inhibitory effect on the constriction of extracted intestines and stomach. 2. In the effect on the ulceration, Souminkwakhyangjeonggisan showed inhibitory effect on the ulcer induced by tying pylorus, while both two prescription showed inhibitory effect on the ulcer induced by Indomethacin. 3. In the effect on the gastric secretion, two prescriptions showed no significant effect on the secretion of gastric juice, while in the effect on pH, both two prescriptions showed significant inhibitory effect on the secretion of gastric acid. And in the effect on Pepsin secretion, only Kwakhyangjeonggisan inhibited Pepsin secretion with statistical significance. 4. In the effect on the movement of small intestine, Kwakhyangjeonggisan decreased the movement of small intestine, while Souminkwakhyangjeonggisan increased the movement of small intestine. And in the effect on the movement of large intestine, both two prescriptions showed inhibitory effect on the movement of large intestine. 5. In the effect on the diarrhea induced by Castor oil, both Kwakhyangjeonggisan and Souminkwakhyangjeonggisan showed inhibitory effect. 6. In the effect on the temperature, Souminkwakhyangjeonggisan showed anti-decreasing effect on normal temperature with statistical significance, while both two prescriptions showed antipyretic effect on mouse fever with statistical significance. Based on these results, it cannot be counted that Kwakhyangjeonggisan has better effect on Oegam(influenza or upper respiratory infectious deisease) than Souminkwakhyangjeonggisan. And in the light of the result that Souminkwakhyangjeonggisan showed better anti-decreasing effect on the normal temperature than Kwakhyangjeonggisan, significant stimulatory effect on the movement of small intestine, old classics discription that Souminkwakhyangjeonggisan has the effect of OnWuGangYin (warming gastrointestinal organs and descending Yin) is reconfirmed in this experimental study.

半夏白朮天麻湯 및 加味半夏白朮天麻湯의 抗疲勞效能에 關한 硏究

柳逢夏,朴東源,柳基遠,申秉澈 慶熙大學校韓醫科大學韓醫學硏究所 1999 慶熙韓醫大論文集 Vol.22 No.1

These studies were conducted to investigate the antifatigue effect of Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang liquid extract on muscle artier swimming exercise. These studies measured the results of fatigue materials in the blood of rats before and after swimming, and compared to control group. The results of these studies were summarized as follows: 1. These studies measured the results of fatigue materials without the destruction of muscle cells. 2. After 30 minutes swimming, the lactate increase 46% on control group, Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang worked on sample group by 37% decreation of the lactate, compare to that of control group. This result means Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang control the lactate accumulation on muscles(p<0.05). 3. Activity level of lactate dehydrogenase was more increased by Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang than control group. This result seems related with low lactate level by Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang(p<0.05). 4. Activity levels of triglycerides and creatine phosphotransferase remarkably increased by swimming, but there was not considerable results by Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang(p<0.05). 5. There were no remarkable changes of glucose level, cholesterol level by the Banhabaekchoolchunmatang and Kamibanhabaekchoolchunmatang nor swimming(p<0.05).