• Asian Pacific Journal of Cancer Prevention
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• 제16권13호
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• pp.5371-5376
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• 2015

Berberine (B1), isolated from stems of Coscinium fenestratum (Goetgh.) Colebr, was used as a principle structure to synthesize three phenolic derivatives: berberrubine (B2) with a single phenolic group, berberrubine chloride (B3) as a chloride counter ion derivative, and 2,3,9,10-tetra-hydroxyberberine chloride (B4) with four phenolic groups, to investigate their direct and indirect antioxidant activities. For DPPH assay, compounds B4, B3, and B2 showed good direct antioxidant activity ($IC_{50}$ values=$10.7{\pm}1.76$, $55.2{\pm}2.24$, and $87.4{\pm}6.65{\mu}M$, respectively) whereas the $IC_{50}$ value of berberine was higher than $500{\mu}M$. Moreover, compound B4 exhibited a better DPPH scavenging activity than BHT as a standard antioxidant ($IC_{50}=72.7{\pm}7.22{\mu}M$) due to the ortho position of hydroxyl groups and its capacity to undergo intramolecular hydrogen bonding. For cytotoxicity assay against human fibrosarcoma cells (HT1080) using MTT reagent, the sequence of $IC_{50}$ value at 7-day treatment stated that B1 < B4 < B2 ($0.44{\pm}0.03$, $2.88{\pm}0.23$, and $6.05{\pm}0.64{\mu}M$, respectively). Berberine derivatives, B2 and B4, showed approximately the same level of CAT expression and significant up-regulation of SOD expression in a dose-dependent manner compared to berberine treatment for 7-day exposure using reverse transcription-polymerase chain reaction (RT-PCR) assays. Our findings show a better direct-antioxidant activity of the derivatives containing phenolic groups than berberine in a cell-free system. For cell-based system, berberine was able to exert better cytotoxic activity than its derivatives. Berberine derivatives containing a single and four phenolic groups showed improved up-regulation of SOD gene expression. Cytotoxic action might not be the main effect of berberine derivatives. Other pharmacological targets of these derivatives should be further investigated to confirm the medical benefit of phenolic groups introduced into the berberine molecule.

• IMMUNE NETWORK
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• 제9권4호
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• pp.115-121
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• 2009

Natural killer (NK) cells play key roles in innate and adaptive immune defenses. NK cell responses are mediated by two major mechanisms: the direct cytolysis of target cells, and immune regulation by production of various cytokines. Many previous reports show that the complex NK cell activation process requires de novo gene expression regulated at both transcriptional and post-transcriptional levels. Specialized un-translated regions (UTR) of mRNAs are the main mechanisms of post-transcriptional regulation. Analysis of posttranscriptional regulation is needed to clearly understand NK cell biology and, furthermore, harness the power of NK cells for therapeutic aims. This review summarizes the current understanding of mRNA metabolism during NK cell activation, focusing primarily on post-transcriptional regulation.

• Archives of Pharmacal Research
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• 제17권5호
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• pp.375-377
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• 1994

Thirteen kinds of naturally occurring or derivatised thiterpenes, reported to have an antitumoral property, were reinvestigated on the basis of their direct cytotoxicity or the inhibitory activity on cell growth against five kinds of cultured human tumor cells, i.e., A-549, SK-OV-3, SK-MEL-2, XF-498 and HCT15, in vitro. Ursonic acid III, betulinic acid VIII, betulonic acid X and glycrrhetinic acid XI were exhibitied a marked inhibition on cell growth.

• 약학회지
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• 제42권5호
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• pp.480-486
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• 1998

Many antitumor and immune modulating components have been isolated from fungal extracts. In this study, the authors isolated the proteoglycans from cultured mycelia of Lentin us lepideus, including especially the acidic polysaccharide fraction, named lepidan. It was obtained by extraction with hot water followed by purification using DEAE cellulose anion exchange. To elucidate antitumor effects against different type of tumor, the proteoglycans were tested on sarcoma 180, C3H MCA clone 16 and P388 leukemia in vivo. Lepidan showed 58.3% of tumor inhibition against solid form of sarcoma 180 and 58.6% against MCA clone 16. But lepidan did not affect life span of mice against P388 leukemia. Also when Lepidan was applicated to MTT assay, it did not show any direct cytotoxicity against various tumor cells in vitro.

• 대한한의학회지
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• 제27권4호
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• pp.30-47
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• 2006

Objective : The present study is aimed to elucidate the effects of Cirsium japonicum var. ussuriense on immunomodulation and the potential as an herbal remedy for cancer treatment. Method : It was performed through measurement of effects Cirsium japonicum var. ussuriense extract (CJE) on NO production, NK cell cytotoxicity and cytokine gene expressions related with macrophage and NK cell activity. Result : 1. CJE did not show any direct cytotoxic effects on 7250, HT1080, Hep G2 and CT-26 cells. 2. CJE activated macrophages partially to product NO and up-regulated gene expressions for iNOS in RAW 264.7 cells. 3. CJE promoted cytotoxicity of NK cells against YAC-1 cells at higher concentration than 200 ${\mu}g/ml$. 4. CJE up-regulated gene expressions for $IL-1{\beta}$, IL-2, iNOS, $IFN-{\gamma}$ and $TNF-{\alpha}$ in mice splenocytes.　５. CJE inhibited lung tumor metastasis induced by CT-26 cell transplantation compared with the control group.　Conclusion : It could be concluded that CJE is an effective herbal drug for immune modulating and anti-cancer treatment by promoting activity of macrophages and NK cells.

• 한국식품위생안전성학회지
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• 제8권3호
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• pp.157-161
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• 1993

본 연구는 ethanol의 reactive metabolite인 acetaldehyde의 일차 배양혈관 평활근 세포에 대한 독성 발현 양식을 규명하기 위한 연구의 일환으로, prostaglandin 합성과 세포독성과의 관련성 여부를 확인하기 위하여 수행되었다. Acetaldehyde는, 일차 배양 혈관 평활근 세포에서의 prostaglandin 합성을 현저히 증가 시켰으며, 이때, cyclooxygenase activity 는 큰 변화없거나 오히려 감소시키는 경향을 보였다. Cyclooxygenase inhibitor 인 indometancin은 acetaldehyde에 의한 LDH release를 현저히 차단시켰으며, aspirin 및 salicylic acid는 전혀 영향을 주지 못했다. Phospholipase $A_2\;(PLA_2)$ inhibitor로 알려져 있는 dexamethasone은 유의적인 세포 독성 경감 작용을 보이지 못하였으며, Lipoxygenase inhibitor 들인 NDGA, propyl gallate 등은 현저한 독성 경감 효과를 보였다. 이상의 결과로부터 acetaldehyde에 의한 일차 배양 혈과 평활근 세포에서의 prostaglandin 합성 증가는 Cell death에 대한 직접적인 원인이 아님을 추론할 수 있었으며, $PLA_2/lipoxygenase$ inhibitor들의 강력한 세포독성 경감 작용으로 미루어 볼 때, acetaldehyde 에 의한 세포독성은 lipoxygenase 대사 산물 혹은 $PLA_2$의 직접 작용에 기인할 가능성을 확인할 수 있었다.

• 동국한의학연구소논문집
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• 제1권
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• pp.209-236
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• 1992

The microculture XTT antiviral assay method is used to quantitate HIV-1 induced cytopathic effects as modulated by test substances. This relatively simple assay facilitated the safe and rapid determination of in vitro antiviral activity of selected chemicals as well as direct cytotoxicity. This experiment also confirmed that this system measures infection and subsequent viral replication in target cells and XTT formazan formations correlated with the accumulation of extracellular virions, as measured by quantitative HIV-1 induced syncytium foramtion. The present results with Glycyrrhizin using this in vitro culture system demonstrated that effective dose, EC50(the concentration at which increases XTT formazan production in infected cultures to 50% of that in untreated, uninfected controls) was 250ml. As comparison, AZT was included in this experiment and demonstrated that EC50 AZT of was 0.05g/ml, approximately 5,000 times more potent than Glycyrrhizin based on EC50 ratio's alone. However, this potency is limited by severe cytotoxicity of AZT, while Glycyrrhizin is approximately 16 times less toxic(IC50 of Glycyrrhizin 800 and AZT 51 g/ml). While AZT's anti-HIV-1 viral activity is mediated by inhibition of reverse transcriptase of the virus, Glycyrrhizin faild to demonstrate any inhibitory activity against reverse transcriptase. Further study is necessary in order to understand the precise mechanisms of Glycyrrhizin action against HIV-1 viruses. Althouth Glycyrrhizin is less effective antiviral agent than AZT, much less toxicity of Glycyrrhizin is desirable in terms of chronic treatment. Combination treatment of AZT and Glycyrrhizin may be therapeutically beneficial. Clinical effectiveness of two drug combination therapy for AIDS patient is unknown at this time. However, this experimental investigation presents the scientific rational basis for such therapeutic approach.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6481-6486
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• 2013

Background: Our previous study demonstrated cytotoxicity of a crude extract from Patrinia heterophylla Bunge (PHEB). In the present study, we aimed to investigate the effects of isovaltrate acetoxyhydrin (IA) isolated from PHEB on the gastric cancer cell SGC-7901, in order to explore a potential treatment for gastric cancer. Methods: MTT assays were employed to determine the effects of IA on cell vitality and proliferation, with monitoring of cell morphology changes and examination of apoptosis with Annexin V-PI staining. Flow cytometry was used to assess cell cycle progression and mitochondrial membrane potential. The activity of caspase 3, 9 was evaluated by spectrophotometry, and the protein levels of Bax, Bcl2 and Cyclin B1 were analyzed with Western blotting of total proteins extracted from cultured cells. Results: The results demonstrated direct toxicity of IA towards SGC-7901 cells. Evidence of apoptosis included blebbing and chromatin condensation. Annexin V-PI assays revealed early apoptosis, involving rapid depolarization of mitochondrial membranes and activity of caspase 3, 9 signaling pathways. Western blotting showed that Bcl2 and Bax proteins was down- and up-regulated, respectively, and cyclin B1 was up-regulated. Cell cycle analysis further indicated that IA could induce G2/M phase arrest in SGC-7901 cells. Conclusions: In conclusion, we believe that IA induces apoptosis of SGC-7901 cells, therefore providing a potential therapeutic agent for treatment of gastric cancer.

• Journal of Ginseng Research
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• 제33권3호
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• pp.177-182
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• 2009

Macrophages play an important role in the first line of immunologic effects against tumor cells. The effects of nonsaponin red ginseng (NSRG) components on macrophage functions like tumoricidal activity, phagocytic activity, and NO production in young (8-weeks-old) and aged (82-weeks-old) male C57BL/6 mice were assessed in vitro, respectively. The treatment of tumor cells (melanoma B16 cells) with the supernatants of NSRG-treated macrophages resulted in an increase of cytotoxicity at 300 $\mu$g/ml in the aged mice, whereas the supernatants did not have a cytotoxic effect in the young mice. It was observed that the supernatants induced the increase of tumor cell proliferation at 150 $\mu$g/ml in the young mice, suggesting that the supernatants contain growth factors rather than cytotoxic molecules. In addition, NSRG alone had a direct cytotoxic effect on the B16 tumor cells. NSRG had no effect on the NO production by the macrophages in the young mice, while it significantly increased the level of NO release in the aged mice. There was no difference in the phagocytic activities of the macrophages by NSRG in both groups of mice. These results suggest that NSRG has differential effects on the macrophage functions in young and aged mice.

• Asian Pacific Journal of Cancer Prevention
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• 제16권1호
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• pp.71-76
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• 2015

Magnesium sulfate is widely used as a food additive and as an orally administered medication. The aim of this study was to evaluate the possible cytotoxicity of magnesium sulfate on AGS human gastric adenocarcinoma cells and gastric mucosa in mice. A trypan blue exclusion assay was used to determine the reduction in viability of AGS cells exposed to magnesium sulfate, and then effects on cell proliferation were quantified. The role of magnesium sulfate-mediated pro-inflammatory cytokine production in AGS cells was also investigated. mRNA expression for IL-$1{\beta}$, IL-6, IL-8, and TNF-${\alpha}$ was determined by RT-PCR, and secretion of these cytokines was measured by ELISA. Immunohistochemical evaluation of IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ expression was conducted in mouse gastric mucosa. Addition of 3 to 50 mM magnesium sulfate to AGS cells inhibited both cell proliferation and cell viability in a dose-dependent manner. Magnesium sulfate had little effect on production of IL-$1{\beta}$ or IL-6 but significantly inhibited production of IL-8. The animal model demonstrated that magnesium sulfate induced production of IL-$1{\beta}$, IL-6, and TNF-${\alpha}$. These preliminary data suggest that magnesium sulfate had a direct effect on the stomach and initiates cytotoxicity in moderate concentrations and time periods by inhibiting viability a nd proliferation of AGS cells and by regulating expression and/or release of pro-inflammatory cytokines.