The present study was designed to explore the antioxidant effect of Bamboo powder and its immunoreactivity in pigs. We investigated the functional properties of Bamboo extracts by means of measuring the contents of total polyphenols and flavonoid as well as determining ABST, DPPH radical scavenging activity, and hydroxyl radical scavenging activity and anticancer activity. The total phenolic compound and flavonoids contents of Bamboo extracts were 171.25 mg/g and 127.5 mg/g, respectively. The DPPH radical, hydroxyl radical, ABST radical scavenging activity of Bamboo extracts were 17.3%, 12.5% and 21.5%, respectively. Evidenced by MTT and cell cycle assay, Bamboo dose-dependently inhibited the cell proliferation and induced G0/G1-phase arrest in CHO cells at concentrations of 100, 250, and 500 ${\mu}g/ml$ Bamboo extracts. More than 80% of apoptotic cells were observed by staining with annexin V in 500 ${\mu}g/ml$ Bamboo-treated CHO cells, indicating that Bamboo had potent anticancer activities. Next, to investigate the effect of Bamboo on cytokine, immunoglobulin concentration, and blood compositions, flatting pigs were fed with Bamboo powder for 38 days. Flatting pigs were divided into 4 groups; basal diet (control), basal diet supplemented with 1% Bamboo powder (T1), 2% Bamboo powder (T2), and 3% Bamboo powder (T3). The level of hemoglobin increased in the all Bamboo-fed groups compared with the normal control group. In particular, platelet levels in the all Bamboo-treated groups increased by approximately 90% compared with the levels from pig on a normal control. Serum levels of immunoglobulins (IgG, IgA) in the pigs fed Bamboo powder were modestly increased, and the interferon-${\gamma}$ level also was strongly increased in 2% or 3% Bamboo-fed groups compared with the levels in control groups. Together, these results demonstrated that Bamboo extracts had an effective capacity of scavenging for ABTS, DPPH, and hydroxyl radicals and showed correlation with potent phenol and flavonoid contents, thus suggesting its antioxidant potential. Moreover, administration of Bamboo in 2~3% improved blood parameters and platelets, and especially immunity-related ones such as IgG, IgA, and interferon-${\gamma}$, leading to be potential feed additives in flatting pigs.
The death ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/ Apo1L is a cytokine that activates apoptosis through cell surface death receptors. TRAIL has sparked growing interest in oncology due to its reported ability to selectively trigger cancer cell death. Euphorbiae humifusae Wind has been used in traditional Oriental medicine as a folk remedy used for the treatment of cancer. However, the mechanism responsible for the anticancer effects of E. humifusae not clearly understood. Here, we show that treatment with subtoxic doses of water extract of E. humifusae (WEEH) in combination with TRAIL induces apoptosis in TRAIL-resistant human gastric carcinoma AGS cells. Combined treatment with WEEH and TRAIL induced chromatin condensation and sub-G1 phase DNA content. These indicators of apoptosis were correlated with the induction of caspase activity that resulted in the cleavage of poly (ADP-ribose) polymerase. Combined treatment also triggered the loss of mitochondrial membrane potential. Furthermore, co-treatment with WEEH and TRAIL down-regulated the protein levels of the anti-apoptotic proteins such as Bcl-2, Bcl-xL, XIAP and cIAP-1. Although more study will be needed to examine the detailed mechanisms, this combined treatment may offer an attractive strategy for safely treating gastric adenocarcinomas and the results provide important new insights into the possible molecular mechanisms of the anticancer activity of E. humifusae.
Hizikia fusiforme is a kind of brown edible seaweed that mainly grows in the temperate seaside areas of the northwest pacific, including Korea, Japan and China, and has been widely used as a health food for hundreds of years. Recently, H. fusiforme has been known to exert pharmacological activities including antioxidant, antimutagenic and anticoagulant activities. However, the molecular mechanisms of H. fusiforme in malignant cells have not been clearly elucidated yet. In this study, the effects of ethyl alcohol extract of H. fusiforme (EAHF) on the anti-proliferative effects of MDA-MB-231 and MCF-7 human breast cancer cells were investigated. EAHF treatment resulted in a concentration-dependent growth inhibition by including apoptosis in MDA-MB-231 cells and G1 phase arrest in MCF-7 cells, which could be proved by MTT assay, DAPI staining, agarose gel electrophoresis and flow cytometry analysis. In MDA-MB-231 cells, the increase in apoptosis induced by EAHF treatment correlated with up-regulation of pro-apoptotic Bax expression. EAHF treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant inhibition of poly (ADP-ribose) polymerase, $\beta$-catenin, phospholipase-${\gamma}1$ protein and DNA fragmentation factor 45/inhibitor of caspase-activated DNase. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. fusiforme.
Eleven functional plant materials were identified via a literature search, and their antioxidant capacity and inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 cells were tested. Yields from hot water extracts of the materials were the highest (52.10%) in Lycii fructus, and the yields from Phellinus linteus were the lowest (5.7%). The yields of another were 14.50-42.47%. Total phenol and flavonoids contents were the highest in P. linteus. The $EC_{50}$ values for DPPH and ABTS radical scavenging activities were lower than $100{\mu}g/ml$ for Salvia miltiorrhiza, whereas the values for P. linteus, Scutellaria baicalensis, and Paeonia lactiflora were $100-200{\mu}g/ml$. The $EC_{50}$ value for the superoxide anion radical scavenging activity of all the extracts was higher than $300{\mu}g/ml$. P. linteus for the reducing power was shown the highest activity. $Fe^{2+}$ chelating activity was the highest in the Morus alba extract. In an MTT assay, the cell viability of the RAW264.7 LPS-exposed cells was above 80% in extracts of $50{\mu}g/ml$ and above 77% in extracts of $100{\mu}g/ml$ in all the plant materials except Acanthopanax sessiliflorum. NO production in the RAW264.7 LPS-exposed cells showed a 12-fold increase compared to the control. The NO production level of all the extracts was $6.86-26.18{\mu}M$. Notably, $100{\mu}g/ml$ of S. baicalensis extract showed a remarkable decrease in NO production (72%) compared with the control. The potent antioxidant and anti-inflammatory activities of S.baicalensis, P. linteus, S. miltiorrhiza, M. alba, and P. lactiflora suggest that they are potential candidates as functional materials.
This study aimed to investigate the suppressive effect of acorn extracts, by evaluating 70% ethanol extract (AE) and hot water extract (AW) using 3T3-L1 preadipocytes. We applied various levels (0, 100, 200, 300 and $500 {\mu}g/mL$) of AE and AW to 3T3-L1 preadipocytes. The cell viability of the 3T3-L1 preadipocytes was not affected by up $300 {\mu}g/mL$ of extracts, but was suppressed by level $500{\mu}g/mL$ of both AE and AW by 20% and 9% respectively. The accumulation of lipid droplets in differentiated 3T3-L1 preadipocytes was dose-dependently suppressed by AE and AW. Especially, at high concentrations ($300{\mu}g/mL$), AE (42%) was more effective than AW (41%). Reactive oxygen species (ROS) was also dose-dependently suppressed by treatment with AE (58%) and AW (52%). With regard to the mRNA related to differentiated 3T3-L1 preadipocytes, $PPAR-{\gamma}$ and aP2 were suppressed by treatment with AE (54 and 40%) and AW (38 and 18%). From our results, acorn extract (AE) has more suppressive effects than AW in differentiated 3T3-L1 preadipocytes. We therefore concluded that acorn has suppressive effects against obesity in differentiated 3T3-L1 cells due to antioxidation.
Kim, Na-Mi;Koo, Bon-Suk;Lee, Seong-Kye;Hwang, Eui-Il;So, Seung-Ho;Do, Jae-Ho
Journal of Ginseng Research
/
v.31
no.2
/
pp.86-92
/
2007
This study was carried out to develop health & functional food by using Korean red ginseng for prevention of skin wrinkles. Effects of Korean red ginseng on the collagen biosynthesis and inhibition of matrix metalloproteinase-I (MMP-1) activity in human dermal fibroblast were investigated. Crude saponin contents of Korean red ginseng water extract (WE), Korean red ginseng ethanol extracts (EE) and Korean Red ginseng purified extracts (PE) were 72 mg/g, 107 mg/g and 220 mg/g, respectively. We incubated human fibroblast cell with Korean red ginseng component by addition of l ${\mu}g/ml$, 5 ${\mu}g/ml$, 10 ${\mu}g/ml$. Amount of collagen biosynthesis was 1.86 ng/ml in control sample and 2.85 ng/ml, 2.05 ng/ml and 2.58 ng/ml in retinoic acid, EE and PE respectively. Furthermore, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were shown 2.01 ng/ml and 3.07 ng/ml. MMP-l activities of EE, PE, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were decreased to 92%, 94%, 91% and 78% respectively as compared with control. Cell proliferation were showed 84-96% in the Korean red ginseng components. The antioxidative SOD activities of the Korean red ginseng components were showed 28-69%, however it was lower than that of Vitamin C. From this results, we conclude that Korean red ginseng have a anti-wrinkle effect and $ginsenoside-Rb_1$ may be considered as a more effective component.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.31
no.3
/
pp.199-218
/
2005
Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.
To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.
Journal of the Korean Society of Food Science and Nutrition
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v.43
no.1
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pp.47-54
/
2014
The aim of this study is to evaluate the effects of green tea polyphenol (GTP) on anticancer treatment with cisplatin (CP), using both an in vitro cell culture model and an in vivo mouse model of established breast tumor. Mouse breast cancer cells (EMT6) were treated with or without GTP and CP followed by determination of the cell viability using an MTT assay. The relative cell viability of CP treated EMT6 cells was 96% at a 20 ${\mu}g/mL$ concentration of cisplatin; however, in combination with GTP (50 ${\mu}g/mL$), the cell viability decreased to 20% at the same concentration of CP (20 ${\mu}g/mL$). For the in vivo study, EMT6 cells were inoculated into Balb/c mice for the establishment of a tumor-bearing mice model. The tumor-bearing mice were treated with CP (5 mg/kg. i.p.) with or without dietary GTP (0.2% drinking water). Tumor growth was monitored by a measurement of tumor size using a digital caliper, and nephrotoxicity was determined by enzymatic and histological examinations. The levels of p53 and caspase-3 in tumor tissues were examined by a Western blot. In tumor-bearing mice treated with GTP plus CP, the increment of tumor volume showed a significant reduction, compared with CP or GTP alone. The levels of p53 and cleaved caspase-3 (caspase-3/p17) in tumor tissues of tumor-bearing mice were increased by CP and GTP compared to CP alone. In CP treated tumor-bearing mice, ${\gamma}$-glutamyltranspeptidase (GGT) and alkaline phosphatase (AP) activities were decreased, and marked tubular necrosis and dilatation were observed in the kidney. CP-induced enzymatic and histopathological changes in the kidney of tumor-bearing mice were reduced by combinations of GTP with CP. The results of these experiments demonstrated that dietary GTP has a potentiating effect on CP anti-tumor activity and a protective effect against CP-induced renal dysfunction. Therefore, GTP may be used as a modulator in anticancer treatment with CP.
Journal of the Korean Society of Food Science and Nutrition
/
v.29
no.1
/
pp.147-152
/
2000
Various lines of evidence suggest that dietary components protect the initiation of carcinogenesis. In this study, the ethanol extracts (AGE) and the methanol and hexane partition layers (AGEM, AGEH) of the Angelica radix were screened for their cytotoxic effects using the MTT assay on HepG2, HeLa, MCF7 and SW626 cells and for their ability to induce quinone reductase (QR) in HepG2 cells. AGEM and AGEH of the Angelica radix showed the strongest cytotoxic effects on HepG2 and HeLa cells. Cell growth was inhibited by 99.8% and 99.8% on HepG2 cells and 99.3% and 99.4% on HeLa cells, at dose of $100\;\mu\textrm{g}/ml$ of AGEM and AGEH extracts respectively. AGE and AGEH significantly induced QR activities in the HepG2 cells. The QR activities of HepG2 cells grown in the presence of AGE, AGEH, and AGEM at the concentration of $50\;\mu\textrm{g}/mL$ were 313.5, 273.3 and 133.3 nmol/min/mg protein, respectively. Therefore, based on these studies, Angelica radix may be developed into a potentially useful cancer chemopreventive agent.
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