• Asian Pacific Journal of Cancer Prevention
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• 제16권10호
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• pp.4311-4316
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• 2015

Hepatocellular carcinoma (HCC) is a primary liver cancer with high global incidence and mortality rates. Current candidate drugs to treat HCC remain lacking and those in use possess undesirable side effects. In this investigation, the antiproliferative effects of dentatin (DTN), a natural coumarin, were evaluated on HepG2 cells and DTN's probable preliminary molecular mechanisms in apoptosis induction were further investigated. DTN significantly (p<0.05) suppressed proliferation of HepG2 cells with an $IC_{50}$ value of $12.0{\mu}g/mL$, without affecting human normal liver cells, WRL-68 ($IC_{50}$ > $50{\mu}g/mL$) causing $G_0/G_1$ cell cycle arrest via apoptosis induction. Caspase colorimetric assays showed markedly increased levels of caspase-3 and caspase-9 activities throughout the treatment period. Western blotting of treated HepG2 cells revealed inhibition of $NF-{\kappa}B$ that triggers the mitochondrial-mediated apoptotic signaling pathway by up-regulating cytoplasmic cytochrome c and Bax, and down-regulating Bcl-2 and Bcl-xL. The current findings suggest DTN has the potential to be developed further as an anticancer compound targeting human HCC.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.629-635
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• 2014

Background: Acute myeloid leukemia (AML) is a fatal hematological malignancy which is resistant to a variety of chemotherapy drugs. Myeloid cell leukemia-1 (Mcl-1), a death-inhibiting protein that regulates apoptosis, has been shown to be overexpressed in numerous malignancies. In addition, it has been demonstrated that the expression level of the Mcl-1 gene increases at the time of leukemic relapse following chemotherapy. The aim of this study was to target Mcl-1 by small interference RNA (siRNA) and analyze its effects on survival and chemosensitivity of acute myeloid leukemia cell line HL-60. Materials and Methods: siRNA transfection was performed with a liposome approach. The expression levels of mRNA and protein were measured by real-time quantitative PCR and Western blot analysis, respectively. Trypan blue assays were performed to evaluate tumor cell growth after siRNA transfection. The cytotoxic effects of Mcl-1 siRNA (siMcl-1) and etoposide were determined using MTT assay on their own and in combination. Apoptosis was quantified using a DNA-histone ELISA assay. Results: Transfection with siMcl-1 significantly suppressed the expression of Mcl-1 mRNA and protein in a time-dependent manner, resulting in strong growth inhibition and spontaneous apoptosis. Surprisingly, pretreatment with siMcl-1 synergistically enhanced the cytotoxic effect of etoposide. Furthermore, Mcl-1 down-regulation significantly increased apoptosis sensitivity to etoposide. No significant biological effects were observed with negative control siRNA treatment. Conclusions: Our results suggest that specific suppression of Mcl-1 by siRNA can effectively induce apoptosis and overcome chemoresistance of leukemic cells. Therefore, siMcl-1 may be a potent adjuvant in leukemia chemotherapy.

• 대한한의학방제학회지
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• 제20권2호
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• pp.65-82
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• 2012

Objectives : The aim of this study was verification of the anti-oxidative and anti-inflammatory effects of Odukhwan(ODH) and Sasinhwan(SSH) in mouse macrophage, RAW264.7 cells. Methods : To investigate the anti-oxidative effect and scavenging activities of DPPH radical, superoxide anions, nitric oxide and peroxynitrite were measured. Cytotoxic activity of extract of ODH and SSH on RAW264.7 cells was measured using MTS assay. To proof the reductive activity of intracellular oxidation, DCFH-DA assay was performed. The nitric oxide(NO) production was measured and pro-inflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2) and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : After those analyses, we bring to a conclusion as follows. Both herbal formulations scavenged DPPH radical and nitric oxide. But ODH had no scavenging activity of superoxide anions and SSH had low scavenging activity in peroxynitrite. And the results indicated that ODH and SSH inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of IL-$1{\beta}$ and IL-6 production in RAW264.7 cells. They also have suppression effects of LPS-induced NF-${\kappa}B$ activation. Conclusions : ODH and SSH have anti-oxidative and anti-inflammatory effects and they may be a part of database for development of new anti-oxidative and anti-inflammatory drugs.

• 대한한의학회지
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• 제25권1호
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• pp.85-95
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• 2004

The theory of liver by Jang suk-sun[張錫純] is that first, although liver exists at right side of body and spleen at left anatomically, function of liver presents at left side of body and function of spleen at right based on principle of 'interdependence between eum and yang' and 'join strength with elasticity', and in the relation between liver and spleen, if gi of spleen ascends, gi of liver also ascends, and if gi of stomach descents, gi of gall bladder also descents. So. care of spleen and stomach becomes main point in treating disease of liver. The meaning of 'the liver being in charge of the evaporation'[肝主氣化] is that first, it raises the primordial gi and forms the 'great g' for it's circulation of whole body. Second., it excretes the functional activity of gi and connects heart with kidney and guides the evaporation of the whole body by connecting innate nature with acquired nature. Third, 'the liver being in charge of the evaporation' is realized by the help of spleen and stomach. And he said that this functional activity of gi is one of distinctive features that distinguish Oriental medicine from Western medicine. He discoursed upon physiology of 'the liver being in charge of the evaporation' and 'the Body belonged to yin and the Use belonged to yang' after seeing an evil of abusing drugs that calm the liver and check hyperfunction of liver by contemporary doctors. And he established a treat of 'warming and recuperating the gi of liver' [溫補肝氣法] and used it for symptoms of 'deficiency of liver-gi'[肝氣虛], 'exhaution of liver-gi'[肝氣脫], and 'the liver-cold'[肝寒證].

• 한국약용작물학회지
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• 제7권4호
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• pp.316-324
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• 1999

중국에서 약용식물은 서양의학과 동등한 수준으로 인간의 건강증진과 유지 및 치료를 위해 매우 중요한 역할을 수행하고 있는데 본 논문에서는 중국에서 약용식물의 산업적 발달에 관해서 간략히 고찰하였다. 현계 중국에서 약용식물로 이용되고 있는 식물은 11, 118종에 달한다. 이들 약용식물이 가공되지 않고 천연산물로 이용되는 연간 총 생산액은 60억달러에 달하며 조제형태로 이용되는 총 생산액은 30억달러에 달한다. 지금까지 약용식물로부터 100여종 이상의 신약이 개발되었으며 천연자원에 의존하려는 세계적인 추세를 감안한다면 약용식물의 중요성은 더욱 증대할 것이며 산업적 개발이 인류의 건강을 유지하는데 결정적 역할을 할 것으로 기대된다.

• 대한한의학회지
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• 제25권3호
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• pp.78-89
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• 2004

Background : A report published by the National Institute of Toxicological Research (NITR) in January 2004 about toxic hepatitis in Korea contained the result of analysis on 55 cases of severe toxic hepatitis from 7 university hospitals for 8 months. NITR claimed that the extrapolated annual frequency of severe toxic hepatitis in Korea was 1904 cases per year. They also claimed that the most frequent etiology of severe toxic hepatitis were herbal medications and similar plant preparations (61.7%), contrasted with traditional therapeutic preparations and healthy foods (29.1%). I have investigated that report to be certain of the result because it is a very important subject for public health and society in Korea. Results : The NITR report has too many problems to have faith in its results. They include the following: 1. The report uses only 55 cases to estimate annual prevalence rate of severe toxic hepatitis in Korea. 2. There was a large regional preponderancy in the NITR report (2 cases in Seoul from a population of 10.17 million, 19 cases in Gwangju from a population of 1.4 million) 3. There was another preponderancy that selected much fewer cases caused by western medication (9.1%) than other reasons. 4. The NITR report used a modified scale than that officially recognized to diagnose toxic hepatitis. 5. There was a mistake using the scale to adapt the right indications. 6. They collected cases before beginning the study, although it was a prospective study. There was also not any questionnaire or other materials concerned with alcohol, drugs, or history of past liver disease. Conclusions : NITR is one of the important official arms of the government of Korea. Nevertheless, there is a severe problem in validity because of selection bias, uncertain accuracy, and insufficiency of raw materials in the report. Therefore it seems incorrect to generalize the results of the report and there is a lack of confidence in it as a national study publishing by the NITR.

• 대한한의학회지
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• 제23권2호
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• pp.19-27
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• 2002

Objective : This study was designed to investigate the effect of Hyeolbuchukeo-tang (HCT) on NO production and the molecular mechanism of NO production modulated by HCT in the primary VSMC (vascular smooth muscle cells). Method : Primary VSMC was established from aorta and cultured VSMC used in this study. NO production of VSMC was assayed by Griess reagent and the expression of iNOS gene was assayed by Western, RT-PCR. Result : $TNF-{\gamma}$ induced NO production, but $IFN-{\gamma}$ or HCT alone did not induce NO production in cultured VSMC. However, $IFN-{\gamma}$ or HCT potentiated NO production in $TNF-{\gamma}-treated$ VSMC in a time- and dose-dependent manner. $TNF-{\gamma}$ induced the iNOS gene expression corresponding to NO production in $TNF-{\gamma}-treated$ VSMC. HCT potentiated NO production in $TNF-{\gamma}-treated$ VSMC by about 20%, but HCT did not increase the level of iNOS mRNA in $TNF-{\gamma}-treated$ VSMC. HCT slightly increased the level of iNOS protein in $TNF-{\gamma}-treated$ VSMC. Calcium ionophore A23187 decreased NO production in $TNF-{\gamma}-treated$ VSMC, but HCT attenuated the effect of A23187. Conclusion : As NO is deeply involved in the development of arteriosclerosis and dilation of blood vessels, drugs or chemicals modulating NO production in VSMC could be used for preventing and treating arteriosclerosis. Considering the effect of HCT on the modulation of NO production in VSMC, MCT has a potential capacity for preventing and treating diseases of the circulation system including arteriosclerosis.

• 대한한의학회지
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• 제30권4호
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• pp.28-36
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• 2009

Objectives: Fatigue is a prevalent symptom encompassing both acute and chronic manifestations. Most fatigue symptoms can be cured by taking a rest or removing underlying causes. However, chronic fatigue is frequently problematic due to its duration and effect on quality of life. There are no particularly effective therapies for chronic fatigue of unknown causes, and patients in Korea usually visit an Oriental clinic. This study aimed to analyze the current status of treatments and patients with chronic fatigue, and then map out of a strategy for development of generalized-treatments for chronic fatigue in Oriental Medicine. Methods: Clinical information related to chronic fatigue was selected from various different databases such as PubMed, KoreaMed, KStudy, DBPIA, OIM, and KOMS. Also, to understand current tendency of medical examination and treatment related with chronic fatigue, we requested Health Insurance Review & Assessment Service for clinical datum from 2003 to 2007. Results: The medical fees of National Health Insurance related with fatigue show an explosive year-on-year increase. On the other hand, it has been decreasing annually in the western medical fields. To take charge of clinical fields related with chronic fatigue by Oriental Medicine, we should make a unified diagnostic system. Then, we should also make standard evaluation tools and develop herbal drugs according to this unified diagnostic system. Conclusions: Fatigue-related symptoms will be a main target of Oriental medicine in the future. We expect that various studies related with chronic fatigue will be undertaken hereafter.

• 대한약침학회지
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• 제17권1호
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• pp.59-69
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• 2014

Objectives: In homeopathy, it is claimed that more homeopathically-diluted potencies render more protective/curative effects against any disease condition. Potentized forms of Condurango are used successfully to treat digestive problems, as well as esophageal and stomach cancers. However, the comparative efficacies of Condurango 6C and 30C, one diluted below and one above Avogadro's limit (lacking original drug molecule), respectively, have not been critically analyzed for their cell-killing (apoptosis) efficacy against lung cancer cells in vitro, and signalling cascades have not been studied. Hence, the present study was undertaken. Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were conducted on H460-non-small-cell lung cancer (NSCLC) cells by using a succussed ethyl alcohol vehicle (placebo) as a control. Studies on cellular morphology, cell cycle regulation, generation of reactive oxygen species (ROS), changes in mitochondrial membrane potential (MMP), and DNA-damage were made, and expressions of related signaling markers were studied. The observations were done in a "blinded" manner. Results: Both Condurango 6C and 30C induced apoptosis via cell cycle arrest at subG0/G1 and altered expressions of certain apoptotic markers significantly in H460 cells. The drugs induced oxidative stress through ROS elevation and MMP depolarization at 18-24 hours. These events presumably activated a caspase-3-mediated signalling cascade, as evidenced by reverse transcriptase-polymerase chain reaction (RT-PCR), western blot and immunofluorescence studies at a late phase (48 hours) in which cells were pushed towards apoptosis. Conclusion: Condurango 30C had greater apoptotic effect than Condurango 6C as claimed in the homeopathic doctrine.

• Biomolecules & Therapeutics
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• 제21권3호
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• pp.190-195
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• 2013

Cisplatin is a member of platinum-containing anti-cancer drugs that causes cross-linking of DNA and ultimately cancer cell apoptosis. The therapeutic function of cisplatin on various types of cancers has been widely reported but the side effects have been discovered together and nephrotoxicity has been regarded as major side effect of cisplatin. To select candidates for new sensitive nephrotoxicity biomarker, we performed proteomic analysis using 2-DE/MALDI-TOF-MS followed by cisplatin treatment in human kidney cell line, HK-2 cells, and compared the results to the gene profile from microarray composed of genes changed in expression by cisplatin from formerly reported article. Annexin A5 has been selected to be the most potential candidate and it has been identified using Western blot, RT-PCR and cell viability assay whether annexin A5 is available to be a sensitive nephrotoxic biomarker. Treatment with cisplatin on HK-2 cells caused the increase of annexin A5 expression in protein and mRNA levels. Over-expression of annexin A5 blocked HK-2 cell proliferation, indicating correlation between annexin A5 and renal cell toxicity. Taken together, these results suggest the possibility of annexin A5 as a new biomarker for cisplatin-mediated nephrotoxicity.