We investigated the chemical components of red onion powder dried using the low temperature vacuum method and the inhibitory effects of solvent extracts of the dried red onion powder on the growth of HT-1080 human fibrosarcoma and HT-29 human colon cancer cells and $H_2O_2$-induced oxidative stress. The moisture content of the dried red onion powder was 17.95%, while the vitamin C content was 96 mg/100 g and the total phenols content was 39.1 mg/mL. The inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts of the red onion powder on the growth of HT-1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was greater on the growth of HT-29 cells, while the A+M extracts had a higher inhibitory effect than the MeOH extracts. Treatment with the hexane, 85% aq. methanol, butanol and water fractions of the extract led to significant inhibition of the growth of both cancer cell lines (p<0.05). Among the fractions, the hexane and 85% aq. methanol fractions showed a greater inhibitory effect. To determine the protective effect on $H_2O_2$-induced oxidative stress, a DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including the crude extracts of dried red onion, appeared to lead to a significant reduction in the levels of intracellular reactive oxygen species (ROS), and these reductions occurred in a dose dependent fashion (p<0.05). Among the fractions, the 85% methanol fraction showed the greatest protective effect on the production of lipid peroxides.
Jung Im Lee;Jung Hwan Oh;Chang-Suk Kong;Youngwan Seo
Journal of Korean Medicine for Obesity Research
/
v.23
no.2
/
pp.69-77
/
2023
Objectives: Atriplex gmelinii C. A. Meyer is a halophyte belonging to the Chenopodiaceae family, and its young leaves and stems are used as fodder for livestock. The aim of the present study was to investigate the effects of A. gmelinii extract and its solvent fractions on lipid accumulation during adipogenesis of 3T3-L1 preadipocytes. Methods: The samples of A. gmelinii were separately extracted using methylene chloride and methanol. Subsequently, they were combined to formulate the initial extract, which was then partitioned based on polarity to prepare solvent fractions. Oil Red O staining was employed to measure lipid accumulation during the differentiation of 3T3-L1 preadipocytes. To verify cytotoxicity in 3T3-L1 cells, MTT assays were conducted. The expression levels of transcription factors in 3T3-L1 preadipocytes were measured through Western blotting analysis. Results: At 50 ㎍/mL, treatment of A. gmelinii extract and its solvent fractions during the differentiation of 3T3-L1 preadipocytes significantly diminished lipid accumulation with no noteworthy cytotoxicity on cell viability. Additionally, when investigating the biochemical pathways that underlie the prevention of lipid accumulation using solvent fractions, it was found that the n-BuOH and n-hexane fractions significantly decreased the expression of key transcription factors involved in the generation of fat, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), and sterol regulatory element-binding protein-1c (SREBP1c). Conclusions: These findings indicate that A. gmelinii can effectively reduce the accumulation of fat in 3T3-L1 adipocytes, making it a potentially valuable material for mitigating and preventing obesity.
Journal of the Korea institute for structural maintenance and inspection
/
v.10
no.4
/
pp.79-86
/
2006
This study was investigated to corrosion resistance of Cr-bearing rebars to macrocell corrosion caused by concrete with crack. Ten types of steel bars having different Cr contents were embedded in concretes with imitation crack. The corrosion resistance of the Cr-bearing rebar was examined by measuring half-cell potential, macrocell corrosion current, corrosion area and weight loss up to 105 cycles of salt spray testing. The results revealed that the Cr content required for corrosion resistance in a macrocell corrosion environment caused by chloride ion gap of $3kg/m^3$ was 9% or more. The corrosion-resisting performance of Cr-bearing rebar was particularly noticeable with a Cr content of 11% or more.
Background and Objectives: Myocardial ischemia-reperfusion injury (MIRI) refers to the damage of cardiac function caused by restoration of blood flow perfusion in ischemic myocardium. However, long non-coding RNA prostate androgen regulated transcript 1 (PART1)'s role in MIRI remain unclear. Methods: Immunofluorescence detected LC3 expression. Intermolecular relationships were verified by dual luciferase reporter assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry and transferase-mediated dUTP nick-end labeling (TUNEL) assays analyzed cell viability and apoptosis. The release of lactate dehydrogenase was tested via enzyme-linked immunosorbent assay (ELISA). Left anterior descending coronary artery surgery induced a MIRI mouse model. Infarct area was detected by 2,3,5-triphenyltetrazolium chloride staining. Hematoxylin and eosin staining examined myocardial injury. ELISA evaluated myocardial marker (creatine kinase MB) level. Results: PART1 was decreased in hypoxia/reoxygenation (H/R) induced AC16 cells and MIRI mice. PART1 upregulation attenuated the increased levels of Bax, beclin-1 and the ratio of LC3II/I, and enhanced the decrease of Bcl-2 and p62 expression in H/R-treated cells. PART1 upregulation alleviated H/R-triggered autophagy and apoptosis via miR-302a-3p. Mechanically, PART1 targeted miR-302a-3p to upregulate transcription factor activating enhancer-binding protein 2C (TFAP2C). TFAP2C silencing reversed the protected effects of miR-302a-3p inhibitor on H/R treated AC16 cells. We further established TFAP2C combined to dual-specificity phosphatase 5 (DUSP5) promoter and activated DUSP5. TFAP2C upregulation suppressed H/R-stimulated autophagy and apoptosis through upregulating DUSP5. Overexpressed PART1 reduced myocardial infarction area and attenuated MIRI in mice. Conclusion: PART1 improved the autophagy and apoptosis in H/R-exposed AC16 cells through miR-302a-3p/TFAP2C/DUSP5 axis, which might provide novel targets for MIRI treatment.
Trichomoniasis is caused by a sexually transmitted flagellate protozoan parasite Trichomonas vaginalis. T. vaginalis-derived secretory products (TvSP) contain lipid mediators such as leukotriene B4 (LTB4) and various cysteinyl leukotrienes (CysLTs) which included LTC4, LTD4, and LTE4. However, the signaling mechanisms by which T. vaginalis-induced CysLTs stimulate interleukin (IL)-8 production in human mast cells remain unclear. In this study, we investigated these mechanisms in human mast cells (HMC-1). Stimulation with TvSP resulted in increased intracellular reactive oxygen species (ROS) generation and NADPH oxidase 2 (NOX2) activation compared to unstimulated cells. Pre-treatment with NOX2 inhibitors such as diphenyleneiodonium chloride (DPI) or apocynin significantly reduced ROS production in TvSP-stimulated HMC-1 cells. Additionally, TvSP stimulation increased NOX2 protein expression and the translocation of p47phox from the cytosol to the membrane. Pretreatment of HMC-1 cells with PI3K or PKC inhibitors reduced TvSP-induced p47phox translocation and ROS generation. Furthermore, NOX2 inhibitors or NOX2 siRNA prevented CREB phosphorylation and IL-8 gene expression or protein secretion induced by TvSP. Pretreatment with a CysLTR antagonist significantly inhibited TvSP-induced ROS production, CREB phosphorylation, and IL-8 production. These results indicate that CysLT-mediated activation of NOX2 plays a crucial role in ROS-dependent IL-8 production in human mast cells stimulated by T. vaginalis-secreted CysLTs. These findings enhance our understanding of the inflammatory response in trichomoniasis and may inform the development of targeted therapies to mitigate this response.
Jo, Seung-Mook;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
Applied Microscopy
/
v.30
no.4
/
pp.357-365
/
2000
Paneth cells have been suggested to contribute to the elimination of excess metals into the intestinal lumen. The purpose of this study wat to investigate the changes of the zinc pools in rats subjected to functional loading with zinc salt by mean of both light and electron microscopical autometallography (AMG). Wistar rats 4 were administrated with zinc chloride (20 mg/kg body weight) intraperitoneally dissolved in 1 ml distilled water. The control group received 1 ml saline IP. After further one hour the animals were transcardially perfused with 0.4% sodium sulphide dissolved in 0.1 M PB fellowed by 3% glutaraldehyde solution for 10 minutes. Pieces of ileum were frozen with solid $CO_2$ and sectioned on a cryostat. The sections $(20{\mu}m)$ were autometallographically developed. Sections selected for EM were reembedded on top of a blank Epon block, from which ultrathin sections (100 nm) were cut. The ultrathin sections were double stained with uranyl acetate (30 min) and lead citrate (5 min), then examined under electron microscope. Studies of comparable sections from control and zinc loaded animals with the AMG selenium method gave quite different results. The control animals demonstrated a weakly positive staining in the cytoplasm of the Paneth cells. In the electron microscope the AMG silver grains were found to be located in the cytoplasm, while the electron dense secretary granules and other cell organelles were void of staining. Few AMG grains were located at the apical surface of the Paneth cells. In sections from zinc loaded rats, the AMG grains were seen in abundance in the lumen of the Lieberkuhn crypts at light microscopic levels. At EM levels the zinc revealing silver grains were located in the cytoplasm as in the controls, but much more AMG grains were shifted into the secretary granules. Furthermore, profound AMG grains were found in the lumen of the crypts and surrounding vessels. And a few grains were seen in the endothelium. The AMG technique demonstrated a pattern of AMG grains in the Paneth cells that strongly suggests a transport of zinc ions through these cells.
Objectives: To evaluate the protective effects of glutathione (GSH) on the cytotoxicity of mercurial compounds$(CM_3HgCl,\;HgCl_2)$ or cadmium chloride$(CdCl_2)$ in EMT-6 cells. Methods: The compounds investigated were $CH_3HgCl,\;HgCl_2,\;CdCl_2$, GSH, buthionine Sulfoximine(BSO), L-2-oxothiazolidine-4-carboxylic acid(OTC). Cytotoxicity analysis consist of nitric oxide(NO) production, ATP production and cell viability. Results: Mercurial compounds and cadmium chloride significantly decreased cell viability and the synthesis of NO and cellular ATP in EMT-6 cells. GSH was not toxic at concentrations of 0-1.6 mM. In the presence of GSH, mercurial compounds and cadmium did not decrease the production of ATP and nitrite in EMT-6 cells. The protective effects of GSH against the cytotoxicity of mercurial compounds and cadmium depended on the concentration of added GSH to the culture medium for EMT-6 cells. We evaluated the effects of intracellular GSH level on mercury- or cadmium-induced cytotoxicity by the pretreatment experiments. Pretreatment of GSH was not changed ${NO_2}^-$ and ATP production, and pretreatment of BSO was decreased in dose and time-dependent manner. Pretreatment of OTC was increased ${NO_2}^-$ and ATP production in dose- and tine-dependent manner. Because intracellular GSH level was increased by OTC pretreatment, the protective effect on mercury- and cadmium-induced cytotoxicity was increased. Conclusions: These results indicated that sulfhydryl compounds had the protective effects against mercury-induced cytotoxicity by the intracellular GSH levels.
Kim, You-Ah;Lee, Jung-Im;Kim, Hae-Jin;Kong, Chang-Suk;Nam, Taek-Jeong;Seo, Young-Wan
Journal of Applied Biological Chemistry
/
v.52
no.4
/
pp.180-186
/
2009
Whole plants of Vitex rotundifolia were extracted for 2 days with methylene chloride ($CH_2Cl_2$) followed by extraction of the residue for an additional 2 days. The same procedure was also applied using methanol (MeOH). The two crude extracts were combined and partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was further partitioned between n-hexane and 85% aq. MeOH, and the aqueous layer was also further fractionated with n-BuOH and $H_2O$, successively. From the 85% aq. MeOH fraction, one compound was isolated through the repeated HPLC. According to the results of physicochemical data including NMR and MS, the chemical structure of the compound was determined as artemetin (1). The antiproliferative effects of the crude extracts, fractions, and compound against HT1080, AGS, MCF-7 and HT-29 human cancer cells were compared with the control by using MTT assay. In the comparative analysis, the 85% aq. MeOH fraction exhibited the strongest antiproliferative effects on human cancer cell lines in a dose-dependent manner (p<0.05). In addition, exposure of compound 1 isolated from 85% aq. MeOH fraction led to strong antiproliferative effect in HT1080 cancer cell lines. These results suggest that the extracts and compound isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents.
Changes of growth and histopathological feature in the mantle structure of the equilateral venus, Gomphina veneriformis exposed to tribultyltin chloride (TBTCl) for 36 weeks were observed. Concentrations of TBTCl were 0, 0.4, 0.6, and $0.8{\mu}g/L$. A regression analysis by power function of SPSS was shown that the growth of experimental groups was significantly decreased after 12 weeks of exposure. For histological analysis, mantle tissues were characterized using H-E stain, AB-PAS (pH 2.5) reaction and Masson's trichrome stain, and epidermal layer thickness and mucous cell distribution were analysed using the image analyser. The mantle had 4-folds (inner-inner, inner-outer, middle, and outer) and its epidermal layer consisted of simple epithlia. A periostracum was observed in the periostracal groove between middle and outer fold. Inner epidermal layer consisted of simple ciliated columnar epithelia, but the outer epidermal layer consisted of simple non-ciliated columnar epithelia. Alcian blue positive mucous cells showed blue color (7462c, 653c) in the inner fold, violet color (2583c) in the middle fold, and blue color (647c, 7455c) in inner epidermal layer (numbers in the parenthesis are codes of Pantone process coated color). Hemolymph sinus in the mantle was extended, and mucous cells in inner plica of the middle fold were stained as blue (7455c) and violet (2587c), after 12 weeks of TBTCI exposure. Cilia and striated border were disappeared, and number of mucous cells in the inner epidermal layer was reduced. Serious histopathological changes in middle and outer fold near the periostracum were observed after 36 weeks. Moreover, epidermal layer thickness and mucous cell distribution were showed decreasing tendency as exposure time to TBTCI was increased. Results of this study suggested that TBTCl induced growth disorder with histopathological changes.
Pathways both mediated by and independent of transferrin(Tf) and the TfR have been described for the accumulation of iron. Although it is not clear whether the same systems take up iron and gallium, these pathways may suggest the contention that uptake of Ga-67 can, in fact, occur by both Tf-independent and Tf-dependent systems and may share with Fe-59 in part the same mechanism for uptake. The predominant system by which uptake of both radiometals occurs may be different in the degree of the transformation of tumor. Transformed(MMSV/3T3) and untransformed(BALB/3T3) cells were incubated with luM of Ga-67-citrate of Fe-59-chloride for 15 min. at $37^{\circ}C$ in either the presence or absence of Tf. After then, the monolayers were washed with HBSS or PBS, and the cells were solubilized in 1% SDS for gamma well counting and protein determinations. There were similarities, as well as differences, in the pattern of uptake of Fe-59 and Ga-67 presented both in ionic from and as bound to Tf. Both radiometals appeared gain to cells in either ionic or Tf-bound forms. Transformed cells appeared to accumulate more radiometal, either Ga-67 or Fe-59 in the presence of Tf than do the their untransforemd counterparts. Conversly the presentation of either radiometal in ionic form resulted in significantly greater accumulation of metal by the untransformed cells than those transformed. The efficiency for uptake of Ga-67 or Fe-59 in the absence of Tf was greater than for uptake of the Ga-Tf or Fe-Tf. However, the magnitude of difference in efficiency of uptake was greater for Fe-59(10-fold) than for Ga-67 (3-fold). Our results Supports the theory that both Tf-independent and Tf-dependent systems for the uptake of Ga-67 both systems operate oppositely between transformed cells and those untransformed, with uptake by the predominating in transformed cells by the Tf-mediated system and in untransformed cells by the Tf-independent. The uptake of Ga-67 by tumor may share with Fe-59 in part the same mechanism.
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