• IMMUNE NETWORK
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• v.11 no.1
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• pp.79-94
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• 2011

Background: Dendritic cell (DC)-based vaccines are currently being evaluated as a novel strategy for tumor vaccination and immunotherapy. However, inducing long-term regression in established tumor-implanted mice is difficult. Here, we show that deoxypohophyllotoxin (DPT) induces maturation and activation of bone marrow-derived DCs via Toll-like receptor (TLR) 4 activation of MAPK and NF-${\kappa}B$. Methods: The phenotypic and functional maturation of DPT-treated DCs was assessed by flow cytometric analysis and cytokine production, respectively. DPT-treated DCs was also used for mixed leukocyte reaction to evaluate T cell-priming capacity and for tumor regression against melanoma. Results: DPT promoted the activation of $CD8^+$ T cells and the Th1 immune response by inducing IL-12 production in DCs. In a B16F10 melanoma-implanted mouse model, we demonstrated that DPT-treated DCs (DPT-DCs) enhance immune priming and regression of an established tumor in vivo. Furthermore, migration of DPT-DCs to the draining lymph nodes was induced via CCR7 upregulation. Mice that received DPT-DCs displayed enhanced antitumor therapeutic efficacy, which was associated with increased IFN-${\gamma}$ production and induction of cytotoxic T lymphocyte activity. Conclusion: These findings strongly suggest that the adjuvant effect of DPT in DC vaccination is associated with the polarization of T effector cells toward a Th1 phenotype and provides a potential therapeutic antitumor immunity.

• Journal of the Korean Chemical Society
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• v.5 no.1
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• pp.33-37
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• 1961

We have shown that carboxy-peptidase destroys the biological activity of angiotensin octa-and deca-peptides. Since Proline occurs as the seventh amino acid from the amino end of the chain and since carboxypeptidase does not cleave proline from a peptid chain, it is evident that the heptapeptid H.asp-arg-val-tyr-ileu-his-pro.OH is formed by this hydrolysis. This peptide must then be biologically inactive. In order to determine whether the phenyl group of the C-terminal amino acid was the necessary requirement for biological activity of the octapeptide, $ala^8$ angiotensin octapeptide(amino acids of peptides numbered from amino end) was synthesized. For this synthesis the four dipeptides were prepared: carbobenzoxy-L-prolyl-L-alanine-P-nitrobenzyl-ester, m.p. $134-135^{\circ}C,$ carbobenzoxy-L-isoleucyl-imidazole benzyl-L-histidine methyl ester, m.p. $114-116^{\circ}C,$ carbobenzoxy-L-valyl-L-tyrosine hydrazide and carbobenzoxy B-benzyl-L-aspartyl-nitro-L-arginine. The first three dipeptides were obtained as crystalline compounds. Imidazole-benzyl-L-histidine was used in the hope that it would block the histidine imidazole against side reactions in steps subsequent to the formation of the C-terminal tetrapeptide. Also, it was through that the imidazole benzylated peptides would be easier to crystallize. This, however, was not the case. The tetrapeptide, carbobenzoxy-L-isoleucyl-L-im, benzyl-histidyl, L-prolyl-L-alanine-nitrobenzyl ester was not obtained in a crystalline form. Neither could the mono-or dihydrobromide of the tetrapeptide free base be induced to crystallize. Carbobenzoxy-L-valyl-L-tyrosine azide was condensed with the tetrapeptide free base to yield the protected hexapeptide; carbobenzoxy-L-valyl-L-tyrosyl-L-isoleucyl-L-im, benzyl, histidyl-L-Prolyl-L-alanine-nitrobenzyl ester. Upon removal of the carbobenzoxy group with hydrogen bromide in acetic acid an amorphous free base hexapeptide ester was obtained. This compound gave the correct C, H, N analysis and contained the six amino acids in the correct ratio. The octapeptide was obtained by condensing this hexapeptide with carbobenzoxy-B-benzyl-L-aspartyl-nitro, L-arginine using the mixed anhydride method of condensation. This amorphous product was proven to be homogenous by chromatography in two solvent systems and upon hydrolysis yielded the eight amino acids in correct ratio. The five protecting groups were removed from the octapeptide by hydrogenolysis over palladium black catalyst. Biological assay of the free peptide indicated that it possessed less than 0.1 per cent of both pressor and oxytocic activity of the phenylalanine8 angiotensin. This suggests that the phenyl group is a point of attachment between angiotensin and its biological receptor site.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.2
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• pp.89-101
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• 2019

In recent times, the number of men with late-onset hypogonadism has increased, and interest on this topic has also increased. This study was conducted to investigate effects of the mixture extract of Fructus amomi Amari, Eucommiae cortex, Bombyx batryticatus on improve late-onset hypogonadism. The experimental subjects consisted of three groups: a control group consisting of 8-week-old male ICR mice that had undergone no treatment, an aging-elicited group (AE group) consisting of 50-week-old ICR male mice that had undergone no treatment, and a Mixed herbal extract treatment group (MT group) consisting of 50-week-old ICR male mice that had undergone the mixture extract of Fructus amomi Amari, Eucommiae cortex, Bombyx batryticatus treatment (0.1 g/kg/day) for 6 months. After the experiment, the mice from all the experimental groups were dissected, and they were analyzed through histochemical and immunohistochemical methods. The mixture extract of Fructus amomi Amari, Eucommiae cortex, Bombyx batryticatus reduces aging-induced cell damage and oxidative stress and increases the secretion of serotonin and B-endorphin in aged mice, and promotes spermatogenesis in seminiferous tubules and reduces apoptosis and oxidative stress, and increases androgen receptor, $17{\beta}-HSD$ and GnRH, increases the ratio of smooth muscle to collagen fibers in the corpus cavernosum, increases eNOS, decreases PDE-5 and oxidative stress in aged mice, so it improves depression, reproductive, sexual problems caused by Late-onset hypogonadism. the mixture extract of Fructus amomi Amari, Eucommiae cortex, Bombyx batryticatus inhibits the induction of osteoporosis by increasing decreased bone matrix distribution due to aging, increasing the activities of OPC and OPN, which are produced in osteoblasts, and decreasing RANKL, MMP-3 activity, increasing OPG activity. It also reduces muscle damage, oxidative stress, inflammation and apoptosis of muscle tissue, and increases Myo-D in the sartorius muscle of aged mice for improving muscle atrophy caused by by Late-onset hypogonadism.

• Journal of Neurogastroenterology and Motility
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• v.26 no.1
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• pp.106-116
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• 2020

Background/Aims Emerging evidence shows that the mechanism of irritable bowel syndrome (IBS) is associated with neurotrophic factors and tight junction proteins (TJPs). It is known that there are sex differences in the pathophysiology of IBS. The aim of the present study is to determine expression levels of neurotrophic factors, TJPs, and cytokines according to IBS subtype and sex. Methods From 59 IBS (33 IBS-constipation, 21 IBS-diarrhea, and 5 IBS-mixed) and 36 control patients, colonic mucosa mRNA expression levels of transient receptor potential vanilloid-1 (TRPV1), nerve growth factor (NGF), glial cell-derived neurotrophic factor (GDNF), and various TJPs were assessed by real-time polymerase chain reaction. Western blot was performed to determine levels of zonular occludens-1 (ZO-1). Serum levels of cytokines were measured by enzyme-linked immunosorbent assay. Results TRPV1, GDNF, and NGF mRNA levels were significantly increased in those with IBS-constipation compared to those in controls (all P < 0.05). However, they showed no significant difference between those with IBS-diarrhea and controls. Expression level of TRPV1 correlated with that of GDNF (r = 0.741, P < 0.001) and NGF (r = 0.935, P < 0.001). ZO-1 RNA expression levels were lower (P = 0.021) in female IBS-diarrhea than those in controls, although they showed no significant differences between male IBS-diarrhea and controls. Serum IL-1β levels in female IBS were significantly higher than those of male IBS, especially in IBS-constipation (P < 0.001). Conclusion Our results suggest that neurotrophic factors and IL-1β are closely related to IBS-constipation and that decrease of ZO-1 is an important factor in female with IBS-diarrhea.

• IMMUNE NETWORK
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• v.22 no.2
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• pp.18.1-18.15
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• 2022

Dysfunction of mitochondrial metabolism is implicated in cellular injury and cell death. While mitochondrial dysfunction is associated with lung injury by lung inflammation, the mechanism by which the impairment of mitochondrial ATP synthesis regulates necroptosis during acute lung injury (ALI) by lung inflammation is unclear. Here, we showed that the impairment of mitochondrial ATP synthesis induces receptor interacting serine/threonine kinase 3 (RIPK3)-dependent necroptosis during lung injury by lung inflammation. We found that the impairment of mitochondrial ATP synthesis by oligomycin, an inhibitor of ATP synthase, resulted in increased lung injury and RIPK3 levels in lung tissues during lung inflammation by LPS in mice. The elevated RIPK3 and RIPK3 phosphorylation levels by oligomycin resulted in high mixed lineage kinase domain-like (MLKL) phosphorylation, the terminal molecule in necroptotic cell death pathway, in lung epithelial cells during lung inflammation. Moreover, the levels of protein in bronchoalveolar lavage fluid (BALF) were increased by the activation of necroptosis via oligomycin during lung inflammation. Furthermore, the levels of ATP5A, a catalytic subunit of the mitochondrial ATP synthase complex for ATP synthesis, were reduced in lung epithelial cells of lung tissues from patients with acute respiratory distress syndrome (ARDS), the most severe form of ALI. The levels of RIPK3, RIPK3 phosphorylation and MLKL phosphorylation were elevated in lung epithelial cells in patients with ARDS. Our results suggest that the impairment of mitochondrial ATP synthesis induces RIPK3-dependent necroptosis in lung epithelial cells during lung injury by lung inflammation.

• Journal of Nutrition and Health
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• v.29 no.6
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• pp.642-650
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• 1996

Apo E polymorphism (e2, e3, e4) was among the first reported genetic polymorphism that explained part of the normal variation in plasma cholesterol concentrations. Both alleles E2 and E4 are significantly more frequent in patients with mixed forms of hyperlipidemia and contribute on the observed differences in CHD risk among different populations. Effects of apo E polymorphism on the distribution of plasma lipid profiles were studied in 105 normolipidemic healthy women. The relative frequencies of common alleles for gene locus of apo E in this study were that E3 allele was 0.848, E4 allels was 0.087, and E2 allele was 0.067. SBP and DBP were slightly more elevated in E2 allele than those in E3 and E4. The pulsation was also significantly (p<0.016) increased by E2 allele with excess body fat % in E2 allele. There were no differences in total-, total HDL-, VLDL+LDL-, VLDL- and LDL cholesterol among the apo E alleles. However, apo E2 allele subject had lower level of total HDL and HDL2 cholesterol (P<0.047) and significantly higher lev디 of HDL3 cholesterol (P<0.05) than those in apo E3 and E4 allele subject. The conclusion is that first, it seems that apo E4-mediated alteration through LDL B/E receptors or E receptors in cholesterol metabolism results in lower plasma TG or remanate particles and in higher levels of VLDL+LDL or LDL. Second, apo E2 allele shows reciprocal effects of E4 on the plasma lipid metabolism, respecitvely. Third, apo E2 allele was more atherogenic than apo E4 because the higher levels of HDL3/HDL2 ratio and atherogenic index[(TC-HDL)/HDL]were criticized.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.8
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• pp.992-997
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• 2008

In this study, we investigated the indirect effect of silk-fibroin on osteoclastic differentiation of RAW264.7 cells. The conditioned medium were collected from MC3T3-E1 osbeoblasts treated with $0.001\;mg/mL{\sim}0.1\;mg/mL$ silk fibroin for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of osteoclastic cytokines in the conditioned medium, the protein expression of osteoprotegerin (OPG) with silk-fibroin was not significantly different. However, the protein expression of interleukin (IL)-$1{\beta}$ was specifically lower in a dose dependent manner. In RAW264.7 cells, the conditioned medium with silk-fibroin inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts in a dose dependent manner. Taken together, we demonstrated that the conditioned medium of silk-fibroin treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with inhibiting selective expression of IL-$1{\beta}$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.333-340
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• 2014

The purpose of this study was to develop functional lotus root bugak with plasma lipid reduction capacity by controlling the color of batter used for bugak preparation. Lotus root, nearly colorless, was selected to observe color effects. Gardeniae fructus (GF), Opuntia ficus-indica var. saboten (OF), and green tea (GT), which are colored yellow, red, and green, respectively, were used as coloring agents. Fermented glutinous rice was prepared naturally during winter season by placing glutinous rice and water (1:2, w/w) together in a crock pot for 7 days. Coloring materials (10%, w/w) were blended with glue made from fermented glutinous rice flour to prepare the batter. Cooked lotus root was then mixed with a 1.1-fold amount of batter (w/w) and dried at room temperature. Lotus root bugak (LRB) is pan-fried with un-roasted sesame oil, which is traditionally used as frying oil in Korea. Low-density lipoprotein receptor knockout ($LDLr^{-/-}$) mice (n=36) were fed an atherogenic diet (AD) containing various types of LRB (10 g%) for 10 weeks. Plasma triglyceride, total cholesterol, and LDL-C concentrations decreased significantly in mice fed LRB prepared with OF batter (OFB) and GT batter (GTB) (P<0.05). Protein expression levels of fatty acid synthase (FAS) and 3-hydroxyl-3-methylglutaryl coenzyme A reductase (HMGCR) in the OFB and GTB groups were suppressed compared with the LRB group (P<0.05). In accordance with the results on FAS and HMGCR expression, sterol regulatory element binding protein-I and II (SREBP-I and II), which are responsible for the regulation of FAS and HMGCR gene expression, respectively, were down-regulated compared to the LRB group (P<0.05). In conclusion, the plasma lipid reduction activities of OFB and GTB could be mediated through down-regulation of FAS and HMGCR mRNA expression via suppression of regulatory molecules, SREBP-I and II, in $LDLr^{-/-}$ mice.