• The Journal of Internal Korean Medicine
• /
• v.22 no.1
• /
• pp.79-85
• /
• 2001

Object : Hepatitis Treatment-tang No.1 has been used for the treatment of Liver disease and Jaundice. Long-term EtOH exposure leads to immunoregulatory and detoxification impairment. This study aimed to determine the relationship between TNF-${\alpha}$ production and expression, and EtOH-induced cytotoxicity on Hep G2 cells. Method : Cells were incubated with EtOH in the presence or absence of HT. The cells were tested after 24 hours and, again, after 48 hours. Cytoviability and TNF-${\alpha}$ release were analyzed by MTT assay and enzyme linked immunosorbent assay (ELISA), respectively. After 24 hours of EtOH exposure, the cytoviability decreased, and the release of TNF-${\alpha}$ was increased. Increased amounts of TNF-${\alpha}$ contribute to EtOH-induced cytotoxicity. The Anti-TNF-${\alpha}$ antibody almost abolished it. Interestingly, EtOH-induced cytotoxicity and TNF-${\alpha}$ production were inhibited by HT. Moreover, when HT was used in combination with the anti-TNF-${\alpha}$ antibody, there was a marked inhibition of EtOH-induced cytotoxicity. Results : These results suggest that HT may prevent the cytotoxicity through partial inhibition of the TNF-${\alpha}$ secretion.

• The Journal of Korean Medicine
• /
• v.27 no.4
• /
• pp.182-190
• /
• 2006

Objective : Based on immunological mechanisms, this study examined whether subcutaneous (s.c.) injection of Clematis mandshurica Maxim. water extract (CMA) has anti-inflammatory effects, and its effect on $TNF-{\alpha}$, IL-1 and IL-10 release from synoviocytes on adjuvant arthritis (AA) in the rat. Methods : Complete Freund's adjuvant was used to induce AA in rats. Synoviocytes were separated by the method of collagenase and DNase digestion Synoviocytes proliferation was assayed by 3-(4, 5 dimethylthiazol 2 yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. $TNF-{\alpha}$, IL-1 and interleukin-10 (IL-10) production of synoviocytes was measured with ELISA. The expression of IL-10 mRNA of synoviocytes was determined using RT PCR. Results : There were significant secondary inflammatory reactions in AA rats, accompanied by the decrease of body and immune organs weight simultaneously. Synoviocytes proliferation of AA rats significantly increased, and the levels of $TNF-{\alpha}$ and IL-1 in supernatants of synoviocytes in AA rats were also elevated compared with the sham group. The administration of CMA (2, 5, 10 mg/kg, s.c.) reduced the above changes significantly. In contrast to $TNF-{\alpha}$ and IL-1, IL-10 production and the level of its mRNA of synoviocytes in AA rats apparently decreased. CMA (2, 5, 10 mg/kg, s.c.) markedly increased IL-10 in synoviocytes at protein and transcription level. Conclusion : The results indicate that CMA has a beneficial effect on rat AA due to modulating inflammatory cytokine production of synoviocytes, which play a crucial role in the pathogenesis of this disease.

• The Journal of Korean Medicine
• /
• v.29 no.1
• /
• pp.47-59
• /
• 2008

Objective : Anti-inflammatory effects of the extract of Lycii Fructus on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells were investigated. Method : In order to assess the cytotoxic effect of Lycii Fructus on the raw 264.7 macrophages 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was performed. Reverse transcription-polymerase chain reaction(RT-PCR) analysis of the mRNA levels of tumor necrosis factor-$\alpha$(TNF-$\alpha$) and inducible nitric oxide synthase(iNOS) was performed in order to provide an estimate of the relative level of expression of these genes. The protein level of the inhibitor of nuclear factor-${\kappa}B(I{\kappa}B)$ and nuclear factor-${\kappa}B$(NF-${\kappa}B$) activity was investigated by Western blot assay. NO production was investigated by NO detection. Result : Lycii Fructus suppressed NO production by inhibiting the LPS-induced expressions of iNOS and TNF-$^-\alpha$ mRNA and iNOS protein in RAW 264.7 macrophage cells. Also, Lycii Fructus suppressed activation of NF-${\kappa}B$ in the nucleus. Conclusion : These results show that the extract of Lycii Fructus has anti-inflammatory effect probably by suppressing iNOS expressions through the down-regulation of NF-${\kappa}B$ binding activity.

• Korean Journal of Pharmacognosy
• /
• v.48 no.1
• /
• pp.18-24
• /
• 2017

This study was conducted to determine the anti-inflammatory effect of essential oil extracted from the wood of Chamaecyparis obtusa Sieb. et Zucc. Endl. (Cupressaceae). The essential oil was extracted from the wood of C. obtusa by hydrodistillation method, and conducted the analysis on the chemical composition of the extracted C. obtusa wood oil through GC-MS. The major constituents of the oil were found to be: ${\alpha}-pinene$ (11.4%), cadinene (5.4%), ${\delta}-cadiene$ (9.0%), ${\tau}-muurolol$ (22.2%), ${\alpha}-cadinol$ (20.8%) etc. We attempted to identify the anti-inflammatory activities of the oil when it is injected in the lipopolysaccharide (LPS)-stimulated RBL-2H3 cells, along with its effects on the secretion of interleukin-4 (IL-4), interleukin-13 (IL-13), ${\beta}-hexosaminidase$. According to the cell viability analysis conducted by MTT assay, the oil in $10^{-7}{\sim}10^{-5}%$concentration showed no effect on the cell viability. After RBL-2H3 cells treated by LPS stimulation were exposed to $10^{-7}%$ concentration of C. obtusa wood oil, the expression levels of IL-4, IL-13 within the cell were observed to remarkably decrease. Also, it was attenuated the release of ${\beta}-hexosaminidase$ from mast cells to a significantly meaningful level. These results suggest that C. obtusa wood oil exerts the anti-inflammatory effect, by regulating the expression of inflammatory cytokines, which is a valuable feature to be highly utilized as the functional materials in the future.

• The Journal of Internal Korean Medicine
• /
• v.38 no.1
• /
• pp.1-9
• /
• 2017

Objective: This study investigated the effect of purified Carthamus tinctorius (C. tinctorius) extracted with a hot water and ethanol method on adipogenic differentiation of mouse bone marrow-derived mesenchymal stromal stem cells (mBMSCs). Methods: The C. tinctorius was extracted using hot water and ethanol. The samples were concentrated by a rotary evaporator and were then dried using a freeze-dryer. The mBMSCs were cultured and maintained in a minimum essential medium eagle alpha (${\alpha}-MEM$) supplemented with 10% FBS and 1% antibiotic antimycotic solution. To induce adipogenic differentiation, the cells were treated with Dulbecco's modified eagle's medium-low glucose (DMEM-LG) containing 1 mg/mL insulin, 1 mM dexamethasone, and 0.5 mM 3-isobutyl-1-methylxanthine. To evaluate the adipogenic differentiation ability, oil-red O staining was performed after adipogenic differentiation for 21 days. The mRNA expression and protein level of adipogenic-related genes were quantified by quantitative real-time PCR and western blotting, respectively. Results: In the results of the MTT assay, no concentrations of C. tinctorius extracts showed toxicity on mBMSCs, so we fixed the treatment concentration of the extract at 100 ng/mL. In oil-red O staining, the water-C. tinctorius extract treatment significantly decreased adipogenic differentiation compared with the control and ethanol extract groups. The water-C. tinctorius extract group in particular showed reduced mRNA and protein expression of Peroxisome proliferator-activated receptor gamma ($Ppar{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/ebp{\alpha}$), which are adipogenic-related transcription factors. Conclusion: These data suggest that extract of C. tinctorius decreased the adipogenic differentiation of mBMSCs, while only water-C. tinctorius extract had an effect on different adipogenesis in mBMSCs. The C. tinctorius will be a useful therapeutic reagent for the prevention of obesity-related diseases such as diabetes, hyperlipidemia, coronary artery disease, and osteoporosis.

• Environmental Analysis Health and Toxicology
• /
• v.29
• /
• pp.10.1-10.6
• /
• 2014

Objectives Cigarette smoking had been recorded as the main cause of impaired endothelium-dependent vasodilation in smokers by reducing nitric oxide (NO), a production of endothelial nitric oxide synthase (eNOS). However, the mechanism of NO impairment via eNOS activity is unclear until now. In this study, cell passage is suggested to be a relevant factor to eNOS expression under cigarette smoking stress. Methods Bovine aortic endothelial cells (BAECs) were chosen as the research subject with passages ranking from 6 to 9 (6P to 9P). After exposure of cigarette smoking extract (CSE) solution, MTT assay and Western blot method were performed to check the cell viability as well as eNOS protein concentration. In these experiments, four concentrations of CSE at 0.5, 1, 2, and 4% were selected for treatment. Results Our results showed that cells almost died at 4% of CSE. Besides, eNOS protein mass had a linear decrease under the increase of CSE concentration. In addition, the effect of CSE on eNOS expression was dissimilar between different passages. Conclusions This study indicated that CSE had effect on both cell viability and eNOS expression. Besides, a reduction in protein mass was matched with the decrease of cell viability due to CSE tress. Last but not least, the response of eNOS protein to different concentration of CSE at different passages was disparate, making the hypothesis about cell passage related inhibition of eNOS caused by CSE solution.

• Microbiology and Biotechnology Letters
• /
• v.37 no.4
• /
• pp.361-364
• /
• 2009

Rahnella aquatilis strain AY2000 produces an anti-yeast substance (AYS), however activity of the AYS tends to be decreased by heat. To investigate whether trehalose can protect AYS activity against heat, comparative studies on the AYS with and without trehalose were performed. After heat treatment at high temperatures ($50^{\circ}C-70^{\circ}C$), the AYS with trehalose had higher activity than the AYS without trehalose had. In case of AYS with trehalose (0.3-0.9M), activity of the AYS could be determined at ranging from $7.8\;{\mu}g/mL$ to $31.3\;{\mu}g/mL$ on S. cerevisiae by MTT method. Consequently, activity of the AYS after heating was well maintained by trehalose.

• Journal of Pharmaceutical Investigation
• /
• v.35 no.1
• /
• pp.17-23
• /
• 2005

Polyethylenimine (PEI) has been used as cationic polymers for efficient gene transfer without the need for endosomolytic agents. Various kinds of PEIs with different molecular weight were tested in order to investigate the effects of the molecular weight of PEI on the transfection efficiency and cell cytotoxicity. The ${\beta}-galactosidase$ expression $(pCMV-{\beta}-gal)$ plasmid was used as a model DNA. Complex formation between PEI and pDNA was assessed by 1% agarose gel electrophoresis method. Particle size and zeta-potential of complexes were determined by electrophoretic light scattering spectrometer. In vitro transfection efficiency was assayed by measuring ${\beta}-galactosidase$ activity. Cell cytotoxicity was determined by MTT assay. Particle sizes of the complexes became smaller on increasing molecular weights of PEI and N/P ratios. Surface potential of complexes was increased as the molecular weight of PEI increased. Transfection efficiency of $pCMV-{\beta}-ga1$ on the HEK 293 cells was greatest with PEI 25 K system but having the lowest cell viability. PEI with high molecular weight showed higher transfection efficiency and cell viability than PEI with low molecular weight.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.22
• /
• pp.10015-10020
• /
• 2014

Background: Fentanyl is used as an analgesic to treat pain in a variety of patients with cancer and recently it has become considered to also act as an antitumor agent. The study present was designed to investigate the effects of fentanyl on colorectal cancer cell growth and plausible mechanisms. Materials and Methods: The human colorectal carcinoma cell line HCT116 was subcutaneously injected into nude mice. The viability of HCT116 was tested by MTT assay, and apoptosis by flow cytometry and caspase-3 activity. The expression of Sirt1 and NF-${\kappa}B$ were evaluated by Western blotting and the levels of Sirt1 and NF-${\kappa}B$ by fluorescence method. SiRNA was used to silence and Ad-Sirt1 to overexpress Sirt1. Results: Our data showed that fentanyl could inhibit tumor growth, with increased expression of Sirt1 and down-regulation of Ac-p65 in tumors. Compared with control cells without treatment, HCT116 cells that were incubated with fentanyl had a higher apoptotic rate. Moreover, fentanyl could increase expression and activity of Sirt1 and inhibitor expression and activity of NF-${\kappa}B$, which might be mechanisms of fentanyl action. Conclusions: Fentanyl increased colorectal carcinoma cell apoptosis by inhibition of NF-${\kappa}B$ activation in a Sirt1-dependent manner.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.6
• /
• pp.2847-2851
• /
• 2012

Purpose: To study the killing effects on osteosarcoma cells of cinobufacini and cisplatin in combination and the related mechanisms so as to explore the chemotherapeutic method with integrated traditional Chinese and Western medicines. Methods: Cinobufacini and cisplatin were applied to OS732 cells singly or jointly and survival rates were measured by MTT assay. Changes in cellular shape were observed with inverted phase contrast and fluorescence microscopy and apoptosis rates were analyzed with flow cytometry (FCM). Immunocytochemistry were used to examine the Fas expression of OS732 cells. Results: The combination of cinobufacini and cisplatin had the effect of up-regulating Fas expression and inducing apoptosis. The survival rate of combined application of 100 ${\mu}g/ml$ cinobufacini and 1 ${\mu}g/ml$ cisplatin on OS-732 cells was significantly lower than with either of the agents alone (p<0.01). Changes in cellular shape and apoptotic rates also indicated the apoptosis-inducing effects of combined application were much enhanced. Conclusion: The combination of cinobufacini and cisplatin demonstrated strong killing effects on OS-732 cells which might be related to up-regulation of Fas expression.