• Journal of Pharmacopuncture
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• v.5 no.1 s.8
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• pp.27-42
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• 2002

Objective: The purpose of this study was to investigate the acute and subacute toxicity and sarcoma- 180 anti-cancer effects of Herbal acupuncture with Triglii Semen in mice and rats. Method: Balble mice were injected intraperitoneally with Triglii semen Herbal acupuncture for $LD_{50}$ and acute toxicity test. Sprague Dawley rats were injected intraperitoneally with Triglii semen Herbal acupuncture for subacute toxicity test. The Triglii semen Herbal acupuncture was injected on Chung-wan(CV12) of mice with S-180 cancer cell line. Results: 1. In acute toxicity test, the $LD_{50}$ value was $7.49{\times}10^3$ml, 0.30ml/kg.2. The body weights of mice treated with Triglii semen Herbal acupuncture increased during the acute toxicity test. 3. In acute toxicity test of serum biochenrical values of mice, total protein was decreased in treatment groups I, 2 and 3, albunrin was decreased in treatment groups 2 and 3 compared to the control group. GOT was increased in treatment group I and Alk. Phosphatase was increased in treatment groups 1,2 and 3 compared to the normal group(p<0.05). 4.ln subacute toxicity test, severe tissue injury was found in lung and liver. 5. In subacute toxicity test, the body weight was decreased in treatment groups I and 2 compared to the normal group and the weight of liver. lung and kidney were increased in treatment groups 1, 2 and 3 compared to the normal group.(p<0.05) 6. In subacute toxicity test, RBC, HGB and HCT were decreased in treatment groups 1 and 2 compared to the normal group. MCV was increased in treatment group1 compared to the normal group, MCH was increased in treatment groups 1 and 2 compared to the control group in complete blood count test.(p<0.05) 7. In subacute toxicity test, total protein was decreased in treatment groups 1 and 2 compared to the nonnal group, BUN was increased in treatment groups 1 and 2 compared to the nonnal group, creatinine and uric acid were decreased in treatment groups 1 and 2 compared to the normal group, glucose was increased in treatment group 2 compared to the nonnal group, triglycelide was decreased in treatment groups I and 2 compared to the normal group, total cholesterol was increased in treatment groups 1 and 2 compared to the control group. GOT was decreased in treatment group 2 compared to the normal and control group, AIk. Phosphatase was increased in treatment group 1 compared to the normal and control group.(p<0.05) 8. Median survival time was 17days in treatment group 2 for S- 180 cancer cell treated with Triglii semen Herbal acupuncture. 9. Natural killer cell activity was insignificant for S-180 cell treated with Triglii semen Herbal acupuncture.(p<0.05) 10. lnterieukin-2 productivity was decreased for S-180 cell treated with Triglii semen Herbal acupuncture compared to the normal and control group.(p<0.05) Conclusion: According to the results, we can conclude Herbal-acupuncture with Triglii semen caused toxicity, and caused no effects in S-180 cancer cell.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

• Pediatric Infection and Vaccine
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• v.12 no.1
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• pp.75-85
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• 2005

Purpose : Fungal infection is one of the important causes of morbidity and mortality in patients with hematologic malignancies. Amphotericin B(ABV) and itraconazole(ITZA) have been used as the standard empirical antifungal therapy in neutropenic patients with acute leukemia who have persistent fever that does not respond to antibiotic therapy. ABV is an antifungal drug associated with side effects such as fever and chills, symptoms which may be mediated by pro-inflammatory cytokines such as interleukin-$1{\beta}$(IL-$1{\beta}$) and tumor necrosis factor-${\alpha}$(TNF-${\alpha}$). We assessed modulation of these pro-inflammatory cytokines as well as the anti-inflammatory cytokines(IL-4, IL-1Ra) by ABV and ITZA. Methods : From March 2004 to February 2005, a total of 30 episodes from acute leukemia patients with febrile neutropenia were analyzed for this study. They were randomly allocated to receive intravenous ABV or ITZA for 14 days. Clinical responses were evaluated at the completion of therapy, and cytokine IL-$1{\beta}$, TNF-${\alpha}$, IL-4, and IL-1Ra were measured for determination to know the correlation between two antifungal agents and inflammatory cytokines. Results : Empirical antifungal agents were given to 37 patients(ABV 20, ITZA 17), and 30 patients(ABV 15, ITZA 15) were evaluable for efficacy. White blood cell and absolute neutrophil count in the group treated with ITZA increased early days of treatment, so the duration of neutropenia in ITZA group is shorter. Serum creatinine level is lower in ITZA group than in ABV group but this is not statistically significant. There was no significant difference in response rate between two groups. The IL-$1{\beta}$ was increased in ABV treatment group and the ratio of IL-1Ra/IL-$1{\beta}$ is markedly decreased in ABV treatment group while increased in ITZA group. Conclusion : ITZA and ABV have at least equivalent efficacy as empirical antifungal therapy in neutropenic children with acute leukemia. However ITZA is associated with significantly less toxicity in clinical and molecular aspects.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.05a
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• pp.17-22
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• 2002

지구상에는 수천종의 버섯류가 자생하고 있어 유전자원으로서의 중요성이 지대할 뿐만 아니라 기능성 식품소재 및 각종 약리 활성을 나타내는 신약개발 소재로도 크게 주목을 받고 있다. 이들 버섯은 균사체의 영양대사로 얻어지는 대사산물이 축적된 자실체의 형태로 나타나는데, 최근에 와서 자실체 및 균사체의 추출물이나 균사체 배양물이 체질개선이나 각종 병의 예방과 치료에 효과가 있는 것으로 밝혀져 건강식품이나 의약품으로서의 용도가 크게 증가하고 있는 실정이다. 특히, 담자균이 생산하는 특정 구조를 갖는 다당류는 오래전부터 종래의 화학요법제와는 달리 숙주내의 면역 기능을 부활하여 소위 면역요법제로서의 항암효과를 나타냄이 알려져왔었다. 현재까지 제약 및 의학적인 방법이 질병의 주된 치료방법으로 이용되어 왔지만 최근에 특정식품의 섭취가 만성질환의 발생을 억제 또는 지연시킨다는 연구 보고가 나오면서부터 만성질환의 치료방법으로서 식이요법을 중요하게 생각하게 되었다. 따라서 새로운 식품소재 및 가공식품의 개발을 통한 성인병 등의 각종 질병예방이 국민보건문제 해결에 필수적이다. 현재 일본 등에서는 표고버섯, 구름버섯 및 치마버섯 유래의 다당체 또는 단백다당체인 lentinan, krestin 또는 PS-K, schizophyllan 및 PSP 등이 실용화되어 높은 가격에 판매되고 있다. 국내에서도 야생 구름버섯 자실체로부터 추출한 단백 다당체인 Copolang(광동제약)이 개발되어 PS-K와 유사하게 암의 치료에 병행 사용되고 있고, 또 강력한 항암활성이 보고된 상황버섯의 균사체 추출물인 단백 다당체가 Mesima-Ex FK(한국신약)라는 상품명으로 암의 치료에 병행 사용되고 있는 것으로 알려지고 있다. 담자균류와 아울러 미생물 유래 다당체는 그 구조와 특성에 있어서 매우 다양함을 지니고 있다. 이러한 미생물 유래 다당류의 공업적 생산과 이용에 대한 연구로서는 Leuconostoc mesenteroides가 생산하는 dextran이 혈장증량제로 개발된 이래 Xanthomonas campestris가 생산하는 pullulan, Zoogloea rgmigera가 생산하는zooglan둥이 대표적인 예로 보고되고 있다. 한편, 미생물 유래 다당류는 구성당, 분자량, 화학적 구조 등과 같은 특성의 차이에 의해 많은 종류가 존재하고 있으며, 다양한 물성 및 유화제, 응고제, gel 형성제, 필름 형성제, 흡착제, 안정제, 접착제 등과 같은 용도로 광범위하게 이용되고 있다. 또한 근래에 들어서는 미생물 유래 다당체가 지니는 항암활성이 확인되어 새로운 의약품으로서의 개발 가능성이 기대된다. 그 밖에도 기존에 알려져 있는 식물 및 해조류 유래의 다당체와는 달리, 발효조를 이용한 연속배양에 의해 공업적 대량 생산이 가능하며, 더욱이 생산된 다당체의 분리 및 회수가 용이하다는 이점을 지니고 있다. 최근에 들어서는 유전공학적 기법을이용한 고생산성 변이균주 및 새로운 기능을 지닌 다당체의 개발에 관한 연구가 보고되고 있는 등 고부가가치를 지닌 새로운 바이오 소재로서의 기능 및 용도 개발에 관한 연구가 활발히 진행되고 있다. 이러한 항암 활성을 나타내는 여러 가지 담자균류중 Agaricus blazei로부터 생산되는 다당체는 고형암 이외에 S형 결장암, 난소암, 유방암, 폐암, 간암 등에 효과가 입증되었고, 천연물질에 의한 암 면역요법으로 각광을 받고 있으며, 항암 및 항virus의 완치율과 저지율에서 현재 여러가지 약효가 있는 버섯중에서도 탁원한 효과가 있는 것으로 증명되고 있다. 이들 다당체는 사이토카인을 생산시켜서 T임파구와 B임파구의 항원 특이적인 면역반응을 활성화시키고, 세포장해성 T세포와 활성화 대식세포의 세포장해 기능을 충진시켜서 암세포를 파괴시킨다. 또한 콜로니 자극인자인 사이토카인을 생산시켜서 면역담당세포의 신생을 촉진시키기도 하며, 암의 화학요법과 방사선 요법으로 저하된 백혈구를 회복시키는 역할을 한다. 따라서 최근의 연구동향은 생산된 다당체의 항암활성을 향상시키고자 하여 배양기간중에 interleukin을 의도적으로 첨가하는 경향이 있다. 이러한 항암 활성을 나타내는 담자균체 유래 다당체는 버섯의 기원에 따라 그 형태에 약간의 차이를 나타내기는 하나 그 기본 형태는 ${\beta}-(1,6)-glucosyl$ 분지를 가진 ${\beta}-(1,3)-glucan$이며, 평균 분자량은 50 ${\sim}$ 200만 정도이다. Agaricus blazei의 원산지인 브라질의 피에다데(Piedade) 지방의 환경조건(산지의 습도는 80%, 낮 기온 $35^{\circ)C$, 밤 기온 $20{\sim}25^{\circ}C$로 대단히 높으며, 정기적으로 열대지방 특유의 소나기가 내리는 지역)에서 볼 수 있듯이 Agaricus blazei의 성장 환경은 매우 까다로운 편이며, 날것으로는 보관이 잘 안되기 때문에 그 재배에 큰 어려움이 있다. 또한, 고체배양에 의해 생산된 버섯 자실체로부터 유기용매 및 열수추출 방법으로 다당체를 생산하는 방법은 균일한 형태의 버섯 자실체를 공급받기가 어렵기 때문에 다당체의 생산 수율이 낮고, 많은 노동력이 요구되는 어려움이 있다. 그러나 액체배양에 의한 다당체 생산의 경우는 고체배양에 의한 다당체 생산에 비해 일정한 조건하에서 배양이 가능하다는 장점이 있으며, 항상 균일한 균사체 및 배양액을 얻을 수 있다. 따라서 원하는 유용물질을 쉽게 획득할 수 있는 장점이 있다.