• 대한바이러스학회지
• /
• 제28권1호
• /
• pp.71-78
• /
• 1998

Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

• 대한한의학회지
• /
• 제17권1호
• /
• pp.111-131
• /
• 1996

For the development of antimetastatic agent 41 kinds of crude drugs were used for the evaluation of inhibitory effect of several crude drugs on cell adhesion of pulmonary cancer cells and platelet aggregation. Results were obtained as follows: 1. Water extracts of crude drugs inhibited cell adhesion of A549 to complex extracelluar matrix over 40 % of contol were Houttuyniae Herba, Mylabris, Rhei Radix et Rhizoma, Meliae Cortex, Ferula Resina, Oldenlandiae diffusae Herba at the higher concentration of $10^{-3}g／ml$ while those inhibiting cell adhesion of Bl6-Fo over 40 % of control were $10^{-5}g／ml$ of Houttuyniae Herba, Aurantii Fructus, Lithospermi Radix, Zedoariae Rhizoma. Prunellae Spica, Foeniculi Fructus, Rbei Radix, Scutellariae Radix, Meliae Cortex, Ferula Resina and Oldenlandiae diffusae Herba. 2. MeOH extracts of crude drugs at the concentration of $4{\times}10^{-4}g／ml$ inhibiting cell adhesion of A549 specifically to single extracelluar matrix over 40 % of control were Lithospermi Radix, Agrimoniae Herba, Rhei Radix and Ferula Resina to collagen I, Houttuyniae Herba, Lithospermi Radix, Bupleuri Radix, Salviae miltiorrhizae Radix, Orostachys Herba, Sappan Lignum, Meliae cortex ferula Resina and Coicis Semen to collagen Ⅳ, Mylabris, Agrimoniae Herba to laminin, Houttuyniae Herba and Meliae Cortex to fibronectin. 3. NeOH extracts of crude drugs at the concentration of $4{\times}10^{-4}g／ml$ inhibiting cell adhesion of B16-Fo specifically to single extracelluar matrix over 60 % of control were Lithospermi Radix, Salviae miltiorrhizae Radix, Meliae Cortex and Ferula Resina to collagen I, Lithospermi Radix, Bupleun Radix, Saiviae miltiorrhizae Radix, Ferula Resina and Acanthopanacis Cortex to collagen Ⅳ, Bupleuri Radix, Orostachys Herba to laminin, Houttuyniae Herba to fibronectin. 4. MeOH extracts of crude drugs inhibiting platelet aggregation over 40% of ADP control were at the concentration of $50{\mu}g/m{\ell}$ of Houttuyniae Herba, Angilicae gigantis Radix, Zedoariae Rhizoma. Coicis Semen and $100{\mu}g/m{\ell}$ of Ferula Resina, Orostachys Herba, Salviae miltiorrhizae Radix, Curcumac Radix, Carthami Flos, Lithospermi Radix, Gleditsiae Spina, Sappan Lignum, Acanthopanacis Cortex. These results suggest that several crude drugs including Ferula Resina, Houttuyniae Herba, Lithospermi Radix and Salviae miltiorrhizae Radix chiefly have more possibility to exert antimetastatic activity and require in vivo antimetastatic study.

• 대한한의학회지
• /
• 제30권4호
• /
• pp.129-141
• /
• 2009

Objective: This study was to investigate the anti-tumor effect, safety, safety, mechanism and metabolizing enzyme of Agrimonia pilosa LEDEB (APL) in female C57B/L mouse tumor (in vivo). Method: First, to evaluate the antitumor activity of APL, we divided the mice into four groups: normal, control, APL50 (50mg/kg), and APL100 (100mg/kg). LLC-obtained American Type Culture Collection was used. LLC had been inoculated to induce tumors. To measure the anti-tumor effect of APL, we calibrated tumor size and weight. To analyze the mechanism of anti-tumor in APL, we used western blotting and to observe metabolizing enzyme in APL we used to real-time PCR. Result: APL50 and APL100 significantly inhibited tumor growth from 12 days after medicine injected. APL did not induce caspase-dependent apoptosis in LLC-bearing mouse tumor. In APL100, it decreased 41% and 71% in CYP2D22 and CYP3A11, respectively. Conclusion: These results suggest that APL has some anti-tumor effects in female C57B/L mouse tumor. APL should be used carefully with other drugs related with CYP2D22 and CYP3A11.

• Archives of Pharmacal Research
• /
• 제29권10호
• /
• pp.859-865
• /
• 2006

With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

• 대한화장품학회:학술대회논문집
• /
• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
• /
• pp.108-109
• /
• 2003

Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol, prunol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ON A are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepatoprotective, anti-inflammatory, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. To clarify the effects of UA and ONA on skin barrier recovery, both flank skin of 8-12 weeks hairless mice were topically treated with samples (2mg/ml) after tape stripping, then measured recovery rate using TEWL on hairless mice. The recovery rate increased in UA and ONA treated groups at 6h more than 20% compared to vehicle treated group (p <0.05). For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to Vehicle group from 1 week without TEWL alteration (p<0.005). EM examination using Ru04 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent (ONA$\geq$UA>Vehicle). LM finding showed that stratum corneum was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Vehicle). Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber increasing by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory experiments were also confirmed in vivo findings. This result suggested that the effects of UA and ONA related to not only skin barrier but also collagen and elastic fibers. Taken together, UA and ONA can be relevant candidates to improve barrier function and pertinent agents for cosmetic applications.

• 한국식품과학회지
• /
• 제42권2호
• /
• pp.233-239
• /
• 2010

본 연구에서는 초고압 및 초음파 공정을 병행하여 실시하는 추출공정을 이용하여 매자나무 수피의 면역활성 및 Sarcoma-180에 의해 유발되는 복수암 및 고형암에 대한 항암활성 증진 효과를 평가하였다. 상기 추출 공정을 통해 매자나무 추출물의 수율이 일반 열수 추출에 비해 1.7배 증가하였으며, 정상세포에 대한 생존율을 80%이상 유지하는 것으로 나타나 독성 저감 효과도 갖는 것으로 확인되었다. 특히, 매자나무의 초고압, 초음파 병행 추출물이 소화기계 암세포주인 AGS와 Caco-2에 대해서 약 70%의 암세포 생육 저해활성을 나타내며, 최고 4 이상의 selectivity를 나타내며 선택적인 암세포 사멸 효과를 나타내었다. 기존의 연구에서 항암성 면역증강제로 사용되고 있는 버섯의 단백 다당체와 관련하여 영지버섯이 효과적인 항암 활성을 나타내며, 특히 위암 세포인 AGS에 대해 50 mg/mL의 고농도의 영지버섯 추출물이 76% 이상의 암세포 생육저해 활성을 나타내었다(24). 이러한 연구 결과와 비교할 때 본 연구의 매자나무 추출물이 갖는 소화기계 암세포 생육저해 활성은 매우 높다고 할 수 있다. 또한 항암성 면역 활성 측정 결과 초고압, 초음파 병행 추출물이 일반 열수 추출 공정이나 초음파, 초고압 단독 추출 공정을 통한 추출물에 비해 nitric oxide 및 IgG의 생성을 증가시키는 것을 확인할 수 있었다. 초고압 및 초음파 추출 공정이 약용작물의 활성성분 용출에 기여하며, 특히 저온 초고압 추출 공정이 매자나무의 주요 활성성분인 berberine의 용출을 증가시킨다는 연구 결과(25,26)에 미루어 볼 때 초고압, 초음파 병행 추출 공정이 매자나무 활성 성분 용출에 효과적인 영향을 미치는 것으로 사료된다. In vivo 실험을 통한 추출 공정별 매자나무 추출물의 항암 활성 평가를 통해서 상기 공정을 통한 매자나무 추출물이 40% 이상의 복수암 유발 마우스의 수명율 연장과 약 75%의 고형암 억제 활성을 나타내며 효과적인 항암활성을 확인할 수 있어, 향후 항암성 면역증강제 및 기능성 항암제로서의 기능성 소재화로서의 활용성이 매우 높을 것으로 사료된다.

• 대한화장품학회지
• /
• 제29권2호
• /
• pp.205-232
• /
• 2003

Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1 mg/ml UA or 0.1-1 mg/ml ONA after tape stripping, and TEWL (Transepidermal water loss) was measured . The recovery rate increased in those UA or ONA treated groups (0.1 mg/ml UA and 0.5 mg/ml ONA) at 6 h more than $20\%$ compared to vehicle treated group (p<0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from f week without TEWL alteration (p<0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent $(ONA{\geq}UA>Vehicle)$. LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Veh). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via $PPAR\;\alpha$. Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either $ONA\;(10{\mu}M)$ or UA $(10{\mu}M)$ for 24h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via $PPAR\;{\alpha}$. Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

• 생명과학회지
• /
• 제29권4호
• /
• pp.484-491
• /
• 2019

프로안토시아니딘(proanthocyanidins)은 식물계에 가장 풍부한 폴리페놀성 화합물로 다양한 고등식물의 뿌리, 잎, 열매, 나무껍질 등에 널리 존재할 뿐만 아니라 이러한 원료로 만들어진 차, 와인, 맥주 등과 같은 식품에도 상당량 함유되어 있다. 세포 및 실험동물을 이용한 다수의 연구보고에 의하면 프로안토시아니딘은 항산화활성 및 면역조절활성, DNA 복구 및 항종양 작용과 같은 인체 건강에 유익한 무수한 효과를 가지고 있는 것으로 밝혀졌다. 면역 세포 중 대식세포(macrophage)는 염증반응을 매개하는 중요한 세포로 외부 병원체 제거에 중요한 역할을 수행하고 있다. 그러나 대식세포가 만성 염증을 유발하고 비만, 당뇨병, 대사 증후군 및 암과 같은 다양한 질병에 관여한다는 것 또한 널리 보고되어왔다. 본 연구에서는 마우스의 대식세포주인 RAW264.7세포를 이용하여 프로안토시아니딘의 항염증활성의 일단이 Heme oxygenase-1 (HO-1)의 유도에 의해서 매개됨을 밝혔다. RAW264.7세포에 프로안토시아니딘을 처리한 결과 세포독성을 보이지 않은 농도에서 HO-1의 발현을 증강시켰다. 또한 프로안토시아니딘의 처리는 HO-1의 발현을 조절하는 핵심 전사인자인 Nrf (nuclear factor-erythroid 2-related factor)-2의 핵으로의 이동을 유의적으로 증가시켰다. 프로안토시아닌딘의 처리는 LPS (lipopolysaccharide)에 의해 유도된 NO (nitric oxide)의 생성 및 iNOS (inducible NO synthase)의 발현과 염증성 사이토카인의 생성 및 발현도 유의적으로 억제 하였다. 이러한 결과는 프로안토시아니딘의 항염증제제로서의 개발 가능성을 제시하는 결과이다.

• 대한화장품학회지
• /
• 제30권2호
• /
• pp.263-278
• /
• 2004

Ursolic acid (UA)와 oleanolic acid (ONA)는 pentacyclic triterpenoid 성분으로 많은 식물들과 의학, 임상용 허브 등에 존재한다. 이런 UA나 ONA는 free acid 형태로 나타나거나, 1개 이상의 당이 연결된 aglycone으로 triterpenoid 배당체를 구성한다. UA와 ONA는 유사한 구조를 가지며 비슷한 약리효과를 나타내는 것으로 알려져 있다. 최근 연구에 의하면, 항종양, 간보호, 항염증, 함암 및 항균역할을 하는 것으로 보고되고 있다. 우리는 급성 장벽손상 및 정상 무모쥐 피부에 미치는 영항에 대한 연구를 했다. UA와 ONA의 피부장벽 회복에 대한 효과를 평가하기 위해서, 8-12주 된 무모쥐를 테이프 스트리핑 한 후, 한쪽 옆구리에 0.01 -0.1mg/mL 농도로 UA 또는 ONA를 국소도포하고 한쪽에는 vehicle만 처치하여 경표피 수분손실(TEWL)량을 측정하였다. UA (0.1mg/mL)와 ONA (0.5mg/mL)를 처리한 그룹의 회복률이 vehicle 처리군에 비해 테이프 스트리핑 후 6 h에서 20% 이상 증가했다.(p < 0.01). 또한 UA와 ONA의 급성장벽손상 회복과 함께 정상 피부장벽 기능에 미치는 영향을 확인하였다. 정상 피부장벽 기능에 대한 효과를 알아보기 위해, 보습력과 경표피 수분손실량을 UA와 ONA (각 2 mg/mL)를 처리한 1주째와 3주째에 측정하였고, 또한 표피와 진피의 상태를 확인하기 위해서 현미경 관찰을 실시하였다. 두 시료를 1주째부터 vehicle 도포군과 비교, 경표피 수분손실 없이 보습력을 증가시켰다(p < 0.005). 전자현미경 사진을 통해 UA와 ONA 도포에 따라 분비되는 층판소체의 증감(ONA$\geq$UA$\geq$vehicle)과 지질이중막 구조 이상 여부를 확인하였다 Light microscopy를 통해 각질층의 두께가 약간 증가함을 보였으며, 특히 표피두께 강화와 편평 현상이 나타났다(UA < ONA < Vehicle). 우리는 또한 UA와 ONA가 PPAR $\alpha$를 통해 표피 각질세포의 분화를 촉진함을 관찰하였다. Western blotting 실험을 통해, 표피 각질세포 분화와 관련된 involucrin, loricrin, filaggrin의 단백질 발현이 최소한 2-3배 이상 증가함을 HaCaT 세포에 UA와 ONA(각 10$\mu$M)를 24 h 처리 후 실험 결과로 확인할 수 있었다. 이런 결과를 토대로 UA와 ONA가 장벽기능 향상뿐 아니라 PPAR $\alpha$를 통한 표피 각질세포 분화를 유도함을 제시할 수 있었다. Masson-trichrome과 elastic fiber 염색법을 통해서, UA와 ONA 도포에 따른 콜라겐섬유의 비후(thickening)와 엘라스틴섬유의 신장(elongation)을 조직 사진으로 확인하였다. 시험관 시험을 통한 콜라겐 및 엘라스틴 합성실험과 엘라스틴 분해효소에 대한 저해능 평가를 통해 진피에 대한 UA와 ONA의 효과를 확인할 수 있었다. 이런 결과들을 토대로 UA와 ONA는 피부장벽기능 유지뿐 아니라, 진피 내 콜라겐섬유와 엘라스틴섬유 합성을 촉진하는 것을 관찰할 수 있었다. 이 결과로부터, UA와 ONA는 장벽기능 및 진피강화에 관여할 수 있는 기능성 화장품으로의 응용에 적절한 후보 물질로 제안할 수 있겠다.