• Tuberculosis and Respiratory Diseases
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• v.68 no.3
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• pp.168-174
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• 2010

Background: Sepsis still has a high mortality rate despite adequate supportive care. Newer therapeutic modalities have been developed but they have generally ended in failure. Recently, insulin was reported to have an anti-inflammatory effect by inhibiting the $I{\kappa}B/NF-{\kappa}B$ pathway, and may have therapeutic potential in sepsis. However, the precise mechanism of the anti-inflammatory effect of insulin is unclear. This study examined the role of insulin in activating $I{\kappa}B/NF-{\kappa}B$ in macrophage. Methods: Raw 264.7 cells, a murine macrophage cell line, were used in this experiment. Western blotting using $I{\kappa}B$ Ab and phosphor-specific $I{\kappa}B$ Ab was performed to evaluate the degradation and phosphorylation of $I{\kappa}B$ cells. For the $I{\kappa}B$ Kinase (IKK) activity, an immune complex kinase assay was performed. The level of interleukin-6 (IL-6) was measured by ELISA to determine the level of proinflammatory cytokine. Results: $I{\kappa}B{\alpha}$ degradation began 30 min after lipopolysaccharide (LPS) treatment. However, an insulin pretreatment suppressed the $I{\kappa}B{\alpha}$ degradation caused by the LPS treatment. The phosphorylation of $I{\kappa}B{\alpha}$ and IKK activity was also inhibited by the insulin pretreatment. Finally, the insulin pretreatment showed a tendency to suppress the induction of IL-6 by LPS. Conclusion: Insulin might have an anti-inflammatory effect though partial inhibition of the $I{\kappa}B/NF{\kappa}B$ pathway in macrophage cell lines.

• The Journal of Korean Medicine
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• v.28 no.2 s.70
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• pp.93-113
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• 2007

Objective : To research Trends of studies about anti-inflammation and pain relief, obesity, and safety of herbal acupuncture therapy by analyzing domestic theses, published since 2001, about herbal acupuncture therapy. Methods : Domestic theses, published since 2001, mentioning anti-inflammation or pain relief, obesity, or safety of herbal acupuncture therapy were reviewed and analyzed. These theses were then classified by university, year, and subject. Results : The following results were obtained in this study. 1. Among published theses related to anti-inflammation effects of herbal-acupuncture, studies about arthritis comprised the most part, 52. In theses relating to causes of arthritis, 16 were about adjuvant, which was the most, followed by Type II collagen, LPS and carrageenan. Blood test, reactions of inflammation and revelation of cytokine and immune cellswere methods for evaluating anti-inflammation effect. The tendency of experimental methods was to focus on molecular biologic method. 2. In theses related to pain relief, many clinical attempts with herbal injection were carried out, and Carthami Flos and Scolopendrawere used most. Observing reduction of pain inducing factor and checking behavioral change were methods for evaluating pain relief. 3. In theses related to obesity, research focused on effects in association with spots on the body suitable for acupuncture. There were also attempts comparing effectiveness between single injections and complex injections. Astraball Radix, Angelica Gigantis Radix, Coicis Semen and Taeumjowetang were used. Evaluation of anti-obesity effects were by weight loss, food efficiency, blood lipid profile and evaluation of liver function. 4. In theses related to safety of herbal-acupuncture, Herba Chelidonii Chaenonelis Fructus, Clematis Florida Thunb, Corydalidis Tuber, Paeoniae Radix, and Carthami Flos which marked 2 theses each were most studied. Methods of evaluating safety were mostly by observing liver and kidney functions based on blood test, and by applying herbal injections to clinical treatment. Conclusion : Herbal acupuncture is being used in various ways associating with its anti-inflammation, pain relief and anti-obesity effect. Studies on efficacy and mechanism of herbal acupuncture are being conducted even at the molecular biology level.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.10
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• pp.1379-1387
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• 2013

In a majority of mammals, male infants have heavier body mass and grow faster than female infants. Accordingly, male offspring nursing requires a much greater maternal energy contribution to lactation. It is possible that the maternal-fetal immunoendocrine dialog plays an important role in female preparation for lactation during pregnancy. Immune system genes are an integral part of gene regulatory networks in lactation and tumor necrosis factor alpha ($TNF{\alpha}$) is a proinflammatory cytokine that also plays an important role in normal mammary gland development. The aim of this study was to evaluate the influence of the sex of calf and/or the -824A/G polymorphism in the promoter region of $TNF{\alpha}$ gene on milk performance traits in Black Pied cattle over the course of lactation. We also studied the allele frequency differences of -824A/G variants across several cattle breeds, which were bred in different climatic conditions. The G allele frequency decreased gradually over the course of lactation events in the Black Pied dairy cattle because of a higher culling rate of cows with the G/G genotype (p<0.001). In contrast to the genotypes A/A and A/G, cows with G/G genotype showed significant variability of milk and milk fat yield subject to sex of delivered calf. Milk yield and milk fat yield were significantly higher in the case of birth of a bull calf than with a heifer calf (p<0.03). The G allele frequency varies from 48% to 58% in Grey Ukrainian and Black Pied cattle to 77% in aboriginal Yakut cattle. Our results suggest that the $TNF{\alpha}$-824A/G gene polymorphism may have an influence on the reproductive efforts of cows over the course of lactation events depending on the sex of progeny. Allocation of resources according to sex of the calf allows optimizing the energy cost of lactation. This may be a probable reason for high G allele frequency in Yakut cattle breeding in extreme environmental conditions. Similarly, the dramatic fall in milk production after birth of a heifer calf increases the probability of culling for the cows with the G/G genotype in animal husbandry.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.10
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• pp.1478-1485
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• 2017

Objective: The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Methods: Eighteen-day-old broiler embryos were injected with $100{\mu}L$ of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Results: Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis $factor-{\alpha}$ factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. Conclusion: These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by attenuating the expression of proinflammatory cytokines and chemokines implicated in disease pathogenesis.

• IMMUNE NETWORK
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• v.6 no.1
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• pp.33-42
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• 2006

Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

• The Journal of Pediatrics of Korean Medicine
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• v.24 no.1
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• pp.9-35
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• 2010

Objectives The purpose of this study is to investigate the effect of SPDJTK(SoPungDoJeokTangKami) and concurrent administration of AJ(Atopy cream, Jawoongo)+SPDJTK on atopic dermatitis-like skin lesions by using in NC/Nga atopic dermatitis mouse induced by BMAC-induced mice. Methods Clinical skin score, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mice were evaluated. Moreover, the cytokine level, total cell number, Immunohistochemical staining and Histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue were used in NC/Nga mice. Results Orally administrated SPDJTK with concurrent administration of SPDJTK and AJ decreased the clinical skin score, total cell number of WBC, eosinophils in blood, serum total IgE & IgG1, IL-5, IL-13, IFN-$\gamma$. Also, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, CD3+CD69+, CD3+CCR3+, CCR3+, CD4+CXCR5+ in ALN, absolute cell number of CD3+CCR3+, CCR3+ in DLN, granulocytes in PBMCs, activation cell number of CD3+CD69+, CCR3+, total cell number of CD3+ T cell in dorsal skin tissue were significantly decreased. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell and mast cell in dermis, amount of Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue, gene expression of IL-5, IL-13 mRNA in ALN, CD4+ Th cell in dorsal skin tissue and CCR3+ eosinophils in ALN were all significantly decreased. However, total number of DLN, absolute number of CD3e+ T cell and CD19+ B cell, absolute number of CD4+, number of Th cell in DLN and gene expression of foxp3 mRNA were significantly increased significantly. Conclusions Concurrent administration of SPDJTK and AJ on atopic dermatitis in NC/Nga atopic dermatitis mouse was very effective treatment for atopic dermatitis.

• The Journal of Internal Korean Medicine
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• v.28 no.2
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• pp.304-320
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• 2007

Objective : To evaluate the effect of Astragalus Membranaceus Extrac (AME) on myelosuppression, activity and immune modulation in 5-fluorouracil (5-FU) treated mice. Method : We carried out complete blood count, histological analysis of bone marrow, and cell colony forming assay for hematopoietic progenitor to evaluate the effect of AME on myelosuppression and conducted swimming test, survival rate, nitric oxide (NO) assay, 51Cr release assay in natural killer cell, mRNA expression of $IL-1{\beta}$, IL-2, IL-4, IL-6, IL-10, $TNF-{\alpht}$, $IFN-{\gamma}$, $TGF-{\beta}$ and GM-CSF in spleen cells to evaluate the effect of AME on quality of life (QOL). Results : AME improved 5-FU induced myelosuppression and peripheral blood count was recovered effectively, had significant efficacy to protect against chemotherapy induced marrow-destruction and on hematopoiesis compared with the control group, improved increase survival rate and the swimming time, had a stimulatory effect on macrophage activation and NK cell activity, and up-regulated cytokine gene transcription (IL-2, IL-6, $IFN-{\gamma}$) in murine immunologic system. Conclusion : We can conclude that AM is an effective herbal agent for improvement of myelosuppression and QOL in 5-FU treated mice.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.3
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• pp.645-661
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• 2009

The water extract of Dohongsamul-Tang(DHSMT) has been traditionally used to stroke and brain injuries in Oriental Medicine. The present study was designed to investigate the effects of DHSMT on the gene expression profile of cerebral infarction by cDNA microarray in photothrombotic ischemia mouse model. Photothrombotic ischemia was induced in stereotactically held male BALB/c mice using rose bengal and cold light. MRI was performed 24 hours after inducing photothrombosis using 1.5 T MRI and 47 mm surface coil to obtain T2-weighted, and contrast-enhanced images. After MRI test, animal was sacrificed and the brain sections were stained for hematoxylin and eosin and immunohistochemistry. MRI and histological analysis revealed that lesion of thrombotic ischemia was well induced in the cortex with the evidence of biological courses of infarction. The target area of thrombotic infarction was 1 mm anterior to bregma and 3 mm lateral to midline with 2 mm in diameter, which were decreased by administration of DHSMT. To assess gene expression pattern of cerebral infarction, mRNA was isolated and reacted with microarray chip(Agilant's DNA Microarray 44K). Scatter and MA plot analysis were performed to clustering of each functional genes. M value [M=log2(R/G), A={log2(R ${\times}$ G)}/2] was between -0.5 and +0.5 with 40% difference. After pretreatment with DHSMT, the expression levels of mRNA of many genes involved in various signaling pathway such as apoptosis, cell cycle, cell proliferation, response to oxidative stress, immune response, angiogenesis, and inflammatory cytokine were markedly inhibited in photothrombotic ischemia lesion compared to the control group. These results suggest that DHSMT prevent ischemic death of brain on photothrombotic ischemia model of mice through modulation of gene expression at the transcriptional level.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.310-318
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• 2010

Dioscorea bulbifera is one of the traditional medicinal herb. It commonly used in the treatment of hematemesis, epistaxis, tuberculous cervical lymphadenitis, laryngitis, acute infectious disease in East Asia. In the present study, we have demonstrated the anti-inflammatory effects of Dioscorea bulbifera MeOH extract (DBME) in macrophage cell line. To investigate mechanism of the anti-inflammatory activity, we examined the effects of the lipopolysaccaride (LPS)-induced production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), pro-inflammatory cytokines and expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), p-inhibitory ${\kappa}B{\alpha}$ (p-$I{\kappa}B{\alpha}$), and nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in a murine macrophage cell line RAW 264.7. The RAW 264.7 cells were cultured in DMEM + serum medium for 24 hrs. After serum starvation for 24 hrs, the cells were treated with DBME 0.03, 0.10, 0.30 mg/$m{\ell}$ for 1 h, followed by stimulation with LPS (1 ${\mu}g/m{\ell}$) for activation of immune response. After treatment, cell viability was measured by MTT assay, and NO production was monitored by measuring the nitrite content in culture medium. The protein band of iNOS, COX-2, p-$I{\kappa}B{\alpha}$, and NF-${\kappa}B$ was determined by immunoblot analysis and levels of cytokine were analyzed by sandwich immunoassays. There were three experimental groups: carrageenan, DBME 0.3, 1.0 g/kg. Rats were administrated either carrageenan (40% PEG) or carrageenan + DBME (0.3, 1.0 g/kg body weight) for 4 days (p.o.). To induce acute paw edema, rats were injected 1% carrageenan (100 ${\mu}{\ell}$/rat, dissolved in sterilized saline). The effect of DBME in the carrageenan-induced rat paw edema. As results, DBME has an inhibitory effect on the production of NO, PGE2, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 and on the expression of iNOS, COX-2, p-$I{\kappa}B{\alpha}$ and translocation of NF-${\kappa}B$ to nuclear from cytosol. In addition, DBME effectively inhibited the increases of paw edema induced by carrageenan treatment in vivo. These results suggest that DBME can inhibit production of pro-inflammatory mediators and might be a useful source for treatment of acute inflammatory disease.

• Journal of Life Science
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• v.24 no.1
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• pp.61-66
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• 2014

The purpose of this study was to investigate the immunomodulatory effects of Alpina officinarum (AO) ethanol extract on immunocompromised mice. The mice were injected intraperitoneally with an immunosuppressive drug, cyclophosphamide, and then administrated orally with 30, 100, and 300 mg/kg of ethanol extract of AO (AO 30, AO 100, and AO 300, respectively). The concentrations of cytokines and immunoglobulins (IgM, IgA, IgG) in serum were measured. The body weight of the mice and spleen cell number of the AO-fed group showed no significant difference compared to a control group. The concentrations of several cytokines, including IL-2, IFN-${\gamma}$, and TGF-${\beta}$, in serum showed a significant increase in the AO 100 group compared to the control and other groups (p<0.05). The IL-4 level showed no significant difference in the experimental groups. The supplementation of AO (30, 100, 300 mg/kg) significantly increased the concentration of IgM (p<0.05). The concentration of IgA was significantly increased in the AO 100 group (p<0.05) compared to the control group. It can be concluded that AO ethanol extract enhances immune function by promoting the production of cytokines and immunoglobulins.