Journal of Physiology & Pathology in Korean Medicine
/
v.32
no.4
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pp.261-270
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2018
The aim of this study was to evaluate the antitumor activities of Typhae pollen (TP) by confirming in vitro cytotoxicity and in vivo anti-tumor and immune-modulatory effect with anti-cachexia effect. The MTT assay is used in HepG2 cell to detect potential cytotoxic activities of aqueous extract of Typhae pollen (TPe). After HepG2 tumor cell implantation, eight mice per groups were assigned to six groups. Three different dosages of TPe (500, 250 and 125 mg/kg) were orally administered in the amount of $10m{\ell}/kg$ and sorafenib also administered 20mg/kg, every day for 35 days from 28 days after the tumor cell implantation. We observed the changes on body weights, tumor volume and weights, lymphatic organ, serum interferon $(IFN)-{\gamma}$ levels, splenocytes and peritoneal NK cell activity, splenic tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, IL-10 contents. Periovarian fat weights, serum IL-6 levels, thicknesses of deposited periovarian adipose tissue and mean diameters were also detected to monitor the tumor-related anticachexic effects. In tumor masses, the immunoreactivities of cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase (cleaved PARP) - apoptotic marks, cyclooxygenase-2 (COX-2), inducible nitric oxide synthases (iNOS) and tumor necrosis factor $(TNF)-{\alpha}$ were additionally observed by immunohistochemistry. The results were compared with sorafenib. Decreases of COX-2 were demonstrated in sorafenib and TPe treated mice and also increases of iNOS in tumor masses were observed in TPe, not in sorafenib. TPe increased periovarian fat pad weights compared with tumor-bearing controls and sorafenib treated mice. TPe showed increases of splenic $TNF-{\alpha}$, IL-10 and $IL-1{\beta}$, serum $IFN-{\gamma}$ and NK cell activities corresponding to increases of spleen weights, lymph node weights and non-atrophic changes of lymph nodes. Our results show oral treatment of TPe 500, 250 and 125 mg/kg has potent in vitro and in vivo antitumor activities through modest cytotoxic effects, immunomodulatory effects and apoptotic activities in HepG2 tumor cells. In addition, TPe can prevent cancer related cachexia.
Kim, Min-Ji;Bae, Nan-Yong;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Choi, Jung-Su;Ahn, Dong-Hyun
Journal of the Korean Society of Food Science and Nutrition
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v.45
no.2
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pp.181-187
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2016
Algae is a potential resource with various biological activities. In this study, the anti-inflammatory effect of Grateloupia imbricata Holmes ethanol extract (GIHEE) from red algae was investigated in LPS-induced RAW 264.7 cells. As a result, reduced secretion of pro-inflammatory cytokines [tumor necrosis factors-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6] and nitric oxide (NO) was observed in a dose-dependent manner. Expression of nuclear factor-kappaB (NF-${\kappa}B$) as well as inducible NO synthase and cyclooxygenase-2 proteins was reduced by GIHEE, suggesting that the anti-inflammatory activity of GIHEE is related to suppression of NF-${\kappa}B$ signaling pathways. In addition, GIHEE reduced phosphorylation of mitogen-activated protein kinases. These results suggest that GIHEE can be used as a potential anti-inflammatory therapeutic.
Osteoarthritis (OA) is a degenerative joint disease characterized by a progressive loss of cartilage. And, increased oxidative stress plays a relevant role in the pathogenesis of OA. Ursodeoxycholic acid (UDCA) is a used drug for liver diseases known for its free radical-scavenging property. The objectives of this study were to investigate the in vivo effects of UDCA on pain severity and cartilage degeneration using an experimental OA model and to explore its mode of actions. OA was induced in rats by intra-articular injection of monosodium iodoacetate (MIA) to the knee. Oral administration UDCA was initiated on the day of MIA injection. Limb nociception was assessed by measuring the paw withdrawal latency and threshold. Samples were analyzed macroscopically and histologically. Immunohistochemistry was used to investigate the expression of interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, nitrotyrosine and inducible nitric oxide synthase (iNOS) in knee joints. UDCA showed an antinociceptive property and attenuated cartilage degeneration. OA rats given oral UDCA significantly exhibited a decreased number of osteoclasts in subchondral bone legion compared with the vehicle-treated OA group. UDCA reduced the expression of IL-$1{\beta}$, IL-6, nitrotyrosine and iNOS in articular cartilage. UDCA treatment significantly attenuated the mRNA expression of matrix metalloproteinase-3 (MMP-3), -13, and ADAMTS5 in IL-$1{\beta}$-stimulated human OA chondrocytes. These results show the inhibitory effects of UDCA on pain production and cartilage degeneration in experimentally induced OA. The chondroprotective properties of UDCA were achieved by suppressing oxidative damage and inhibiting catabolic factors that are implicated in the pathogenesis of cartilage damage in OA.
Journal of the Korean Society of Food Science and Nutrition
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v.46
no.6
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pp.681-687
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2017
This study was focused on the anti-inflammatory activities of Annona muricata leaf ethanol extracts (AME). Inflammation of macrophages was induced by lipopolysaccharide (LPS) treatment, and various inflammation-mediated factors [cytokines and nitric oxide (NO)] were measured. AME treatment significantly reduced LPS-induced NO, cytokine levels [interleukin (IL)-6, tumor necrosis $factor-{\alpha}$ and $IL-1{\beta}$], and expression of inducible NO synthase and cyclooxygenase-2 in a dose-dependent manner. Mechanical studies showed that AME treatment inhibited activation of mitogen-activated protein kinase and nuclear factor $(NF)-{\kappa}B$ in macrophages treated with LPS. From these results, AME treatment strongly inhibits LPS-induced inflammation through inhibition of $NF-{\kappa}B$ activation, suggesting AME could be a potential candidate for treatment of inflammatory disease as a nutraceutical drug.
Eupatorium japonicum belongs to a family of Asteraceae plants and flowers of E. japonicum have been consumed as a tea. In this study, we investigated whether E. japonicum extract inhibits lipopolysaccharide (LPS)-induced inflammatory responses in Raw264.7 macrophages. The cells were treated with various concentrations (0, 1, 2.5, 5, or 10 mg/L) of 70% ethanol extract from E. japonicum flowers (EJE) in Raw264.7 cells. LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production were inhibited by EJE up to 67% and 49% of these productions, respectively without any reduction of viable cell numbers. EJE reduced LPS-induced expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 proteins and their corresponding mRNA levels. Additionally, EJE decreased the levels of interleukin (IL)-6, IL-1${\beta}$, and tumor necrosis factor (TNF)-${\alpha}$ mRNA. EJE was further fractionated with water, butanol, ethylacetate (EA), hexane, or methylene chloride (MC). Among the resulting five fractions, EA and MC, respectively from EJE significantly inhibited LPS-induced NO production (each inhibition rate was 85.3% of 10 mg/L EA fraction and 97.2% of 10 mg/L MC fraction) without significant cytotoxicity in Raw264.7 cells. These results indicate that EJE exhibits powerful effects of anti-inflammation and can be developed as a potential anti-inflammatory agent.
Kim, Sung Hwan;Lee, In Chul;Ko, Je Won;Moon, Changjong;Kim, Sung Ho;Shin, In Sik;Seo, Young Won;Kim, Hyoung Chin;Kim, Jong Choon
Biomolecules & Therapeutics
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v.23
no.2
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pp.180-188
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2015
This study investigated the possible effects and molecular mechanisms of diallyl disulfide (DADS) against cyclophosphamide (CP)-induced hemorrhagic cystitis (HC) in rats. Inflammation response was assessed by histopathology and serum cytokines levels. We determined the protein expressions of nuclear transcription factor kappa-B (NF-${\kappa}B$), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), oxidative stress, urinary nitrite-nitrate, malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Finally, we studied the involvement of mitogen-activated protein kinases (MAPKs) signaling in the protective effects of DADS against CP-induced HC. CP treatment caused a HC which was evidenced by an increase in histopathological changes, proinflammatory cytokines levels, urinary nitrite-nitrate level, and the protein expression of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-c-Jun N-terminal kinase (JNK), and p-extracellular signal regulated kinase (ERK). The significant decreases in glutathione content and glutathione-S-transferase and glutathione reductase activities, and the significant increase in MDA content and urinary MDA and 8-OHdG levels indicated that CP-induced bladder injury was mediated through oxidative DNA damage. In contrast, DADS pretreatment attenuated CP-induced HC, including histopathological lesion, serum cytokines levels, oxidative damage, and urinary oxidative DNA damage. DADS also caused significantly decreased the protein expressions of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-JNK, and p-ERK. These results indicate that DADS prevents CP-induced HC and that the protective effects of DADS may be due to its ability to regulate proinflammatory cytokines production by inhibition of NF-${\kappa}B$ and MAPKs expressions, and its potent anti-oxidative capability through reduction of oxidative DNA damage in the bladder.
Objective : Pamooltang (PM) and Sipjeondaebotang (SC) are used in Korea to treat many diseases such as sterility, menstrual disorder, and general prostration. We made a comparative study of PM and SC which are different in component ratio between Astragalus membranaceus BUNGE (AC) and Cinnamomum cassia PRESL. (CC). Methods : Anti-oxidation was studied by 1.1.-diphenyl-2-picrylhydrazyl (DPPH) assay and anti-inflammation was investigated by prostaglandin $E_2\;(PGE_2)$ and nitric oxide (NO) inhibition assay. For immune response activities, this study used NO synthesis on RAW 264.7 cells and splenocyte proliferation. Results : The results showed that PM and SC components had no significant effect of anti-oxidation or anti-inflammation. However, we observed their effects upon inducible NO synthesis in Raw 264.7 cells. The SC2 stimulated NO synthesis $11.42\pm1.36{\mu}M$ (control; $0.89\pm0.00{\mu}M$). PM and SC components had the effect of immune response which in a dose-dependent manner significantly induced the splenocyte proliferation. The splenocyte proliferation induced by SC2 was higher than others at the concentration of 1, 10, 100, 1000 and 2000 ${\mu}g/ml$. The SC8 was shown to up-regulate IgG, 100 ${\mu}g/ml$ 3.3 times, 1000 ${\mu}g/ml$ 2.6 times as a control. Conclusions : These results may have important implications for our understanding of the ratios of AC and CC in SC.
The hemodynamic changes in septic patients produced by inhalational anesthetics are sufficient to threaten the anesthesiologists. The effect of hydroxocobalamin, a vitamin $B_{12a}$, on contractile responses to phenylephrine during administration of inhalational anesthetics were evaluated in aortic ring preparations obtained from LPS-treated rats. The sepsis was developed by intraperitoneal injection of LPS (1.5 mg/kg for l8h) and confirmed by iNOS expression using RT-PCR. Statistical significances (P<0.05) were analyzed by Student's t-test or paired t-test according to data characteristics. The blood pressure, but not heart rate, was decreased in LPS-treated rats as compared to control rats. The contractile response to phenylephrine were dose-dependently increased from the doses of $10^{-8}\;M$ to that of $10^{-5}$ and were attenuated in LPS-treated rings. Both halothane and enflurane, at the doses of 1 MAC, decreased the contractile responses to phenylephrine while isoflurane did not significantly affect the contractile responses. Hydroxocobalamin ($10^{-5}$ M) significantly potentiated the contractile responses in the LPS-treated aortic ring preparations during administration of each inhalational anesthetic or not. From these results, it is suggested that hydroxocobalamin may improve the hemodynamics of septic patients during inhalational anesthesia. Abbreviations: LPS, lipopolysaccharide; RT-PCR, reverse transcription-polymerase chain reaction; MAC, minimum alveolar concentration; iNOS, inducible nitric oxide synthase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase
Objective: The aim of this study was to evaluate the effects of compound organic acid calcium (COAC) on growth performance, hepatic antioxidant status and intestinal barrier of male broilers under high ambient temperature (32.7℃). Methods: Nine hundred healthy one-d-old Cobb-500 male broiler chicks were randomly assigned into three groups with six replicates of 50 birds each. A basal diet supplemented with 0% (control), 0.4% and 0.8% COAC, respectively were fed to birds for 6 weeks. All treatments were under high ambient indoor temperature of 32.7℃, and had a constant calcium and available phosphorus ratio. Results: The results showed that, compared with control, the average daily gain of broilers in 0.4% and 0.8% was significantly increased and the ratio of feed to gain in in 0.4% and 0.8% was significantly decreased at 1 to 21, 22 to 42 and 1 to 42 days of age (p<0.05). Compared with control, 0.8% COAC slightly decreased (p = 0.093) the content of malondialdehyde in liver at 42 days of age while 0.4% COAC significantly decreased (p<0.05) the activity of alkaline phosphatase. Furthermore, 0.4% COAC significantly enhanced the intestinal barrier function via increasing jejunal and ileal ocln transcription, promoting jejunal mucin 2 transcription at 42 days of age (p<0.05), and decreasing jejunal toll-like receptor 2 (TLR-2) and ileal TLR-15, inducible nitric oxide synthase compared with control group (p<0.05). Whereas, no significant differences on the transcription of interleukin-1β in jejunum and ileum were observed among three treatments (p>0.05). Overall, heat stress caused by high natural environment temperature may induce the damage to hepatic antioxidation and intestinal barrier. Conclusion: Dietary inclusion of COAC can improve the tolerance of broilers to thermal environment through the modification of antioxidative parameters in liver and the mRNA expression of genes in intestinal barrier, resulting in an optimal inclusion level of 0.4%.
Lee, M.T.;Lin, W.C.;Lin, L.J.;Wang, S.Y.;Chang, S.C.;Lee, T.T.
Asian-Australasian Journal of Animal Sciences
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v.33
no.7
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pp.1167-1179
/
2020
Objective: This study was conducted to fathom the underlying mechanisms of nutrition intervention and redox sensitive transcription factors regulated by Antrodia cinnamomea fermented product (FAC) dietary supplementation in broiler chickens. Methods: Four hundreds d-old broilers (41±0.5 g/bird) assigned to 5 groups were examined after consuming control diet, or control diet replaced with 5% wheat bran (WB), 10% WB, 5% FAC, and 10% FAC. Liver mRNA expression of antioxidant, inflammatory and lipid metabolism pathways were analyzed. Prostaglandin E2 (PGE2) concentration in each group were tested in the chicken peripheral blood mononuclear cells (cPBMCs) of 35-d old broilers to represent the stress level of the chickens. Furthermore, these cells were stimulated with 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) and lipopolysaccharide (LPS) to evaluate the cell stress tolerance by measuring cell viability and oxidative species. Results: Heme oxygenase-1, glutathione S-transferase, glutamate-cysteine ligase, catalytic subunit, and superoxide dismutase, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) that regulates the above antioxidant genes were all up-regulated significantly in FAC groups. Reactive oxygen species modulator protein 1 and NADPH oxygenase 1 were both rather down-regulated in 10% FAC group as comparison with two WB groups. Despite expressing higher level than control group, birds receiving diet containing FAC had significantly lower expression level in nuclear factor-kappa B (NF-κB) and other genes (inducible nitric oxide synthase, tumor necrosis factor-α, interleukin-1β, nucleotide-binding domain, leucine-richcontaining family, pyrin domain-containing-3, and cyclooxygenase 2) involving in inflammatory pathways. Additionally, except for 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase that showed relatively higher in both groups, the WB, lipoprotein lipase, Acetyl-CoA carboxylase, fatty acid synthase, fatty acid binding protein, fatty acid desaturase 2 and peroxisome proliferator-activated receptor alpha genes were expressed at higher levels in 10% FAC group. In support of above results, promoted Nrf2 and inhibited NF-κB nuclear translocation in chicken liver were found in FAC containing groups. H2O2 and NO levels induced by LPS and AAPH in cPBMCs were compromised in FAC containing diet. In 35-d-old birds, PGE2 production in cPBMCs was also suppressed by the FAC diet. Conclusion: FAC may promote Nrf2 antioxidant pathway and positively regulate lipid metabolism, both are potential inhibitor of NF-κB inflammatory pathway.
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