BACKGROUND/OBJECFTIVES: Artemisinin, a natural product isolated from Gaeddongssuk (artemisia annua L.) and its main active derivative, dihydroartemisinin (DHA), have long been used as antimalarial drugs. Recent studies reported that artemisinin is efficacious for curing diseases, including cancers, and for improving the immune system. Many researchers have shown the therapeutic effects of artemisinin on tumors such as breast cancer, liver cancer and kidney cancer, but there is still insufficient data regarding glioblastoma (GBM). Glioblastoma accounts for 12-15% of brain cancer, and the median survival is less than a year, despite medical treatments such as surgery, radiation therapy, and chemotherapy. In this study, we investigated the anti-cancer effects of DHA and transferrin against glioblastoma (glioblastoma multiforme, GBM). MATERIALS/METHODS: This study was performed through in vitro experiments using C6 cells. The toxicity dependence of DHA and transferrin (TF) on time and concentration was analyzed by MTT assay and cell cycle assay. Observations of cellular morphology were recorded with an optical microscope and color digital camera. The anti-cancer mechanism of DHA and TF against GBM were studied by flow cytometry with Annexin V and caspase 3/7. RESULTS: MTT assay revealed that TF enhanced the cytotoxicity of DHA against C6 cells. An Annexin V immune-precipitation assay showed that the percentages of apoptosis of cells treated with TF, DHA alone, DHA in combination with TF, and the control group were $7.15{\pm}4.15%$, $34.3{\pm}5.15%$, $66.42{\pm}5.98%$, and $1.2{\pm}0.15%$, respectively. The results of the Annexin V assay were consistent with those of the MTT assay. DHA induced apoptosis in C6 cells through DNA damage, and TF enhanced the effects of DHA. CONCLUSION: The results of this study demonstrated that DHA, the derivative of the active ingredient in Gaeddongssuk, is effective against GBM, apparently via inhibition of cancer cell proliferation by a pharmacological effect. The role of transferrin as an allosteric activator in the GBM therapeutic efficacy of DHA was also confirmed.
A total of eight strains of avian reoviruses were isolated from chickens with arthritis or stunted growth. The isolations were made from broilers or broiler breeders under 12 weeks of age. The viruses had a typical morphology of reoviruses with double capsid layers and 81nm of diameter. In agar gel precipitation tests, the isolates reacted with antisera prepared against S-1133 or R-1 strains of avian reoviruses and cross reacted with S-1133 antigen. They did not agglutinated RBC's from day-old chicks, adult chickens, guinea pigs, and horses. The isolates showed strong resistance against the treatments of chloroform, IUdR, and heat, When infectivities of the viruses were titrated in cell cultures of chicken embryo fibroblast, chicken embryo liver, and Vero cells, similar end points reached four to five days after inoculation, regardless of tell types and virus inoculation time, either inoculated simultaneously at the time of cell seeding or on confluency. Mean times of mortality of chicken embryos inoculated with the isolates via the chorioallantoic membrane ranged from 54 to 59 hours and that of S-1133 strain was 73 hours.
Ji, Hwan-Sung;Choi, Jung Hwa;Choi, Kwang Ho;Yoon, Sang Chul;Lee, Dong Woo;Kim, Jin-Koo
Korean Journal of Fisheries and Aquatic Sciences
/
v.47
no.6
/
pp.888-894
/
2014
Seventeen specimens of leptocephali [9.8-44.5 mm total length (TL)], of the family Ophichthidae, were collected from southeastern waters off Jeju Island and the Korea-Japan intermediate zone, and identified by means of morphology and genetics. These specimens were identified as belonging to the subfamily Ophichthinae based on various combinations of morphological characters: 211-215 total myomeres; 7 gut swellings; 2 liver lobes connected with the gall bladder on the second lobe; 6-7 postanal pigments present from anus to caudal margin. An analysis of the partial 12S rRNA sequences (849 base pairs) of mitochondrial DNA showed that our specimens must be Ophisurus macrorhynchos because their sequences were concordant with those of the adult O. macrorhynchos (genetic distance = 0.000). Furthermore, their total myomeres were consistent with those of the O. macrorhynchos adult. This is the first time that the morphological characteristics of O. macrorhynchos leptocephali have been described for Korean waters, and we suggest diagnostic characteristics for the genus Ophisurus leptocephali. We hypothesize that one of the spawning grounds of O. macrorhynchos is located in the southeastern part of Jeju Island.
Objective: Endometrial fibrosis, the primary pathological feature of intrauterine adhesion, may lead to disruption of endometrial tissue structure, menstrual abnormalities, infertility, and recurrent pregnancy loss. At present, no ideal therapeutic strategy exists for this fibrotic disease. Eupatilin, a major pharmacologically active flavone from Artemisia, has been previously reported to act as a potent inducer of dedifferentiation of fibrotic tissue in the liver and lung. However, the effects of eupatilin on endometrial fibrosis have not yet been investigated. In this study, we present the first report on the impact of eupatilin treatment on transforming growth factor beta (TGF-β)-induced endometrial fibrosis. Methods: The efficacy of eupatilin on TGF-β-induced endometrial fibrosis was assessed by examining changes in morphology and the expression levels of fibrosis markers using immunofluorescence staining and quantitative real-time reverse-transcription polymerase chain reaction. Results: Eupatilin treatment significantly reduced the fibrotic activity of TGF-β-induced endometrial fibrosis in Ishikawa cells, which displayed more circular shapes and formed more colonies. Additionally, the effects of eupatilin on fibrotic markers including alpha-smooth muscle actin, hypoxia-inducible factor 1 alpha, collagen type I alpha 1 chain, and matrix metalloproteinase-2, were evaluated in TGF-β-induced endometrial fibrosis. The expression of these markers was highly upregulated by TGF-β pretreatment and recovered to the levels of control cells in response to eupatilin treatment. Conclusion: Our findings suggest that suppression of TGF-β-induced signaling by eupatilin might be an effective therapeutic strategy for the treatment of endometrial fibrosis.
Kim, Ho-Hyun;Bang, Hyui-Jeng;Gang, Yun-Ho;Park, In-Sick;Ahn, Sang-Hyun;Kim, Jin-Tack;Lee, Hai-Poong
Journal of Oriental Physiology
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v.14
no.2
s.20
/
pp.215-224
/
1999
After Triton WR-1339 (TX; 600mg/kg) intraperitoneal injection, hepatic tissues of ICR mice were intragastric injected with Hyulboochucketang extract(HCE; 3.3ml/kg/day) were observed to investigate the suppressive effect of lipid accumulation that evoke by the antihyperlipidemic effect of HCE. These hepatic tissues were fixed in fromol-calcium solution and were cryocut. These tissues stained by H&E for general morphology, sudan black B for lipid and perchloric acid-naphthoquinone(PAN) method for cholesterol. After TX treatment, the increase of hepatocyte having meshlike cytoplasm(HHMC) were shown in all hepatic lobules and the hepatic plates were disappeared in the aggregative region of HHMC. The number of blue black colored lipid drop and dark green colored asterisk shaped cholesterol particle in hepatic cytoplasm were increased and the size of lipid drop and cholesterol particle were enlarged. But, in HCE-treated mice, the HHCM were disappeared and hapatic plate were rearranged. The number of lipid drop and cholesterol particle were decreased than TX-treated mice and the size of lipid drop and cholesterol particle were diminished. As results indicated that the HCE work on the suppression of lipid accumulation in hepatic tissue of hyperlipidemic mice caused by disturbance of lipid metabolism.
BACKGROUND/OBJECTIVES: The $NAD^+$ precursor nicotinamide riboside (NR) is a type of vitamin $B_3$ found in cow's milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced obesity, Alzheimer's disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on inflammation and mitochondrial biogenesis in AML12 mouse hepatocytes. MATERIALS/METHODS: A subset of hepatocytes was treated with palmitic acid (PA; $250{\mu}M$) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR ($10{\mu}M$ and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed. RESULTS: Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and, tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor ${\gamma}$ coactivator-$1{\alpha}$, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes. CONCLUSIONS: These data demonstrated that NR attenuated hepatic inflammation and increased levels of mitochondrial markers in hepatocytes.
Oketch, Elijah Ogola;Lee, Jung Woo;Yu, Myunghwan;Hong, Jun Seon;Kim, Yu Bin;Nawarathne, Shan Randima;Chiu, Josh Wen-Cheng;Heo, Jung Min
Animal Bioscience
/
v.35
no.12
/
pp.1929-1939
/
2022
Objective: To investigate the physiological effects of exogenous emulsifiers in broiler chickens that were fed tallow-incorporated reduced-energy diets over 35 days. Methods: A total of 256 Ross 308 one-day-old broilers (42.28±0.16 g) were randomly allocated in a 2×2 factorial arrangement to 32 pens with eight chicks per cage. Birds were fed one of four dietary treatments as follows: i) positive control (PCN; energy sufficient diet); ii) negative control (NCN; energy-deficient diet, -100 ME kcal/kg); iii) PCL (PCN plus 0.05% emulsifier); and iv) NCL (NCN plus 0.05% emulsifier). Growth performance was evaluated weekly whereas assessments for the carcass traits, digestibility, some blood metabolites, ileal morphology, and meat quality were measured on d 21 and d 35. Results: Birds fed the NCL diet had higher (p<0.05) body weights, daily gains, daily feed intake, and improved feed efficiency over the entire 35-day period. Improvements (p<0.05) for the ileal digestibility of crude fat, energy, and dry matter commensurate with longer (p<0.05) villus heights were also observed with emulsifiers in the NCL and PCL diets. For the carcass measurements, only the liver weights were increased (p<0.05) with emulsifiers in the supplemented groups. For blood metabolites, higher (p<0.05) lipase levels were noticed with emulsifiers in the NCL and PCL diets. In addition, marginal reductions (p = 0.076; p = 0.095, respectively) were also noted with emulsifiers for the total cholesterol and triglyceride contents on d 35. Regarding meat quality, breast muscle yellowness was increased (p<0.05) with emulsifier use in supplemented groups. Conclusion: Our results suggest that emulsifier supplementation at 0.05% in diets could potentially improve the growth performance and nutrient digestibility of broilers over 35 days. This could compensate for the lower growth performance that could be recorded with fat-incorporated lower-energy diets.
Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.
Journal of the Korean Society of Food Science and Nutrition
/
v.39
no.11
/
pp.1595-1603
/
2010
The present study was conducted to investigate the effect of dietary grape pomace on lipid metabolism and hepatic morphology of rats fed a high fat diet. The high fat diet contained additional 15% lard to AIN 93-based diet. Male Sprague-Dawley rats were fed experimental diets containing 5% grape pomace for 4 weeks. Serum activities of glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) were not changed by high fat and grape pomace feeding. Serum concentration of triglyceride in rats fed a high fat diet was decreased significantly by dietary grape pomace. Hepatic concentrations of total lipid, total cholesterol and triglyceride were reduced in grape pomace groups with a high fat diet. Fecal concentrations of total cholesterol and triglyceride were increased in grape pomace groups with a high fat diet. The fecal content of coprostanol was not different among the groups. Dietary grape pomace increased the fecal excretion of cholesterol and coprostanone in rats fed a high fat diet. The fecal excretion of bile acid was not affected by feeding grape pomace in rats fed a high fat diet. Light micrographs of liver tissue revealed lipid droplets were increased by a high fat diet, but dietary supplementation of grape pomace tended to alleviate such changes.
We have recently discovered that mycelium of Phellinus linteus, popular medical mushrooms in Korea, possess alcohol dehydrogenase and produce alcohol. In the present study, it was examined that the effect of fermented rice wine made by using mycelium of P. linteus (FLMP) on the expression of in-flammation-related proteins in both $HepG_2$ cells and rats. To examine the effect of FLMP on the morphology and expression of inflammatory proteins in $HepG_2$ cells, the cells were incubated with ethanol, and FLMP for 24 hours, and then analyzed by microscopic observation and Western blot and reverse transcription polymerase chain reaction (RT-PCR). While ethanol induced the morphological change accompanied with cell debris formation and scattering on $HepG_2$ cells, FLMP had no effect. The results of Western blot and RT-PCR analyses showed that the level of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-1 and COX-2 was induced by ethanol, however, FLMP inhibited the expression of these proteins and its mRNAs. In the animal model, the value of flutamate oxaloacetate transaminase and glutamate pyruvate transaminase was significantly increased by administration with ethanol. But the group administrated with FLMP showed lower levels on the changes of these markers compared with ethanol-administrated group. Besides, the results of Western blot and RT-PCR analyses showed that the expression of inflammatory proteins such as iNOS, COX-1 and COX-2 was not affected by FLMP administration in rat liver. About histopathological and immunohistochemical observations, inflammatory loci were markedly decreased in the FLMP-administrated rat compared to ethanol-administrated rats and showed weaker COX-2 and iNOS jmmunoreactions. These results suggested that FLMP showed slight changes on the inflammatory proteins expression compared to ethanol and FLMP may be used as a functional alcoholic beverage.
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