• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.7
• /
• pp.912-919
• /
• 2017

Objective: Identification of the candidate genes that play key roles in phenotypic variations can provide new information about evolution and positive selection. Interleukin (IL)-32 is involved in many biological processes, however, its role for the immune response against various diseases in mammals is poorly understood. Therefore, the current investigation was performed for the better understanding of the molecular evolution and the positive selection of single nucleotide polymorphisms in IL-32 gene. Methods: By using fixation index ($F_{ST}$) based method, IL-32 (9375) gene was found to be outlier and under significant positive selection with the provisional combined allocation of mean heterozygosity and $F_{ST}$. Using nucleotide sequences of 11 mammalian species from National Center for Biotechnology Information database, the evolutionary selection of IL-32 gene was determined using Maximum likelihood model method, through four models (M1a, M2a, M7, and M8) in Codeml program of phylogenetic analysis by maximum liklihood. Results: IL-32 is detected under positive selection using the $F_{ST}$ simulations method. The phylogenetic tree revealed that goat IL-32 was in close resemblance with sheep IL-32. The coding nucleotide sequences were compared among 11 species and it was found that the goat IL-32 gene shared identity with sheep (96.54%), bison (91.97%), camel (58.39%), cat (56.59%), buffalo (56.50%), human (56.13%), dog (50.97%), horse (54.04%), and rabbit (53.41%) respectively. Conclusion: This study provides evidence for IL-32 gene as under significant positive selection in goat.

• Journal of Acupuncture Research
• /
• v.22 no.2
• /
• pp.1-12
• /
• 2005

Objective: With the onset of stroke, white blood cells release several proinflammatory cytokines, including interleukin (IL)-6, IL-10, and tumor necrosis factor $(TNF)-{\alpha}$. It has been proven in previous studies that the release of these cytokines is related to the extent of damage to the brain and to overall prognosis. However, no studies have yet been performed to determine the connection with IL-6 and IL-10. Thus, this study is performed to see whether polymorphisms of IL-6, IL-10, and $TNF-{\alpha}$ genes that show increased serum concentration with the onset of stroke are related to stroke attack in Koreans. Methods : Peripheral blood samples derived from patients with stroke (n=100) and healthy controls (n=100) were taken under informed consent. In subjects with stroke, blood samples were obtained within 24 hours of stroke onset. Genomic DNA was isolated using the Wizard DNA Purification Kit (Promega, Madison, WI). Results : 1. Subjects with Heterozygote (GA) and Homozygote (AA) $TNF-{\alpha}$ gene types showed 2.433 and 20.457 times higher risks of being attacked by stroke, respectively, compared to subjects with wild type (GG) $TNF-{\alpha}$ gene type. The data was still statistically significant after adjusting for age, sex, history of smoking, and history of alcohol drinking. 2. Subjects with Homozygote (CC) IL-6 gene type showed 182.033 times higher risk of being attacked by stroke, compared to subjects with wild type (GG) IL-6 genes. This data was statistically insignificant (p=0.700). The data was still statistically insignificant after adjusting for age, sex, history of smoking, and history of alcohol drinking. 3. Subjects with Heterozygote (GA) and Homozygote (GG) IL-10 gene types showed 8.785 and 3.303 times higher risks of being attacked by stroke, respectively, compared to subjects with wild type (AA) IL-10 genes. The data was still statistically insignificant after adjusting for age, sex, history of smoking, and history of alcohol drinking. Conclusion : Our results suggest that the investigated $TNF-{\alpha}$ and IL-10 gene polymorphisms play an important role in stroke attack, but IL-6 gene polymorphism has not been found to associated with stroke.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2003.11a
• /
• pp.96-96
• /
• 2003

Previously, wogonin (5,7-dihydroxy-8-methoxyflavone) was found to suppress proinflammatory enzyme expression including cyclooxygenase-2 (COX-2), contributing to in vivo anti-inflammatory activity against skin inflammation. However, the detailed effect on each skin cell type has not been understood. Therefore, present investigation was carried out to find the effect of wogonin on inflammation-associated gene expression from skin fibroblasts in culture using reverse transcriptase-polymerase chain reaction. As a result, it was found that wogonin (10 - 100 ${\mu}$M) clearly down-regulated COX -2 expression from NIH/3T3 cells treated with 12-O-tetradecanoylphorbol 13-acetate, interleukin-1${\beta}$ or tumor necrosis factor-a. But, the expression levels of COX-1, interleukin-1${\beta}$ and fibronectin were not significantly affected. This finding was well correlated with significant reduction of prostaglandin E$_2$(PGE$_2$) production by wogonin. As a comparison, NS-398 (selective cyclooxygenase-2 inhibitor) did not suppress COX -2 expression and other gene levels, while PGE$_2$production was potently reduced at 0.1 - 10 ${\mu}$M. All these results suggest that COX -2 down-regulation of skin fibroblasts may be, at least in part, one of anti-inflammatory mechanisms of wogonin against skin inflammation.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.7
• /
• pp.2951-2957
• /
• 2014

Background: Gastric carcinogenesis is a complicated process that involves environmental and genetic factors like interleukin-4 (IL-4) and IL-8. Single nucleotide polymorphisms in their genes are associated with changed levels of gene expression. Here, we investigated the association between IL4-590 C>T and IL8-251T>A and gastric cancer (GC) risk in Sichuan of Southwestern China. Materials and Methods: We surveyed the research subjects using a self-designed questionnaire with questions on demographic factors and putative risk factors. Approximately 2-5ml of whole blood was collected after field survey to analyze IL4-590 C>T and IL8-251T>A genotypes using MALDI-TOF MS. Results: Our study recruited 308 pairs of GC patients and controls, including 224 (72.7%) men and 84 (27.3%) women in each group. There were 99 cardia and 176 noncardia GC patients in the case group. The case and control groups had an average age of $57.7{\pm}10.6$ ($mean{\pm}SD$) and $57.6{\pm}11.1$ years. GC patients reported a significantly greater proportion of family history of cancer (29.9% vs 10.7%, p<0.01) and drinking (54.6% vs 43.2%, p<0.01) than did controls. Variant genotypes of IL-4-590 C>T and IL-8-251 T>A were not associated with overall GC risk (adjusted OR, 0.89; 95%CI, 0.61-1.28 for CT or CC vs TT; adjusted OR, 1.14; 95%CI, 0.86-1.79 for TA or AA vs TT). Stratification analysis of two SNPs for risk by subsites only found that variant IL-8-251 TA or AA genotype was associated with increased noncardia GC risk (adjusted OR, 2.58; 95%CI, 1.19-5.57). We did not observe interactions between the IL-8-251 T>A genotype and smoking (adjusted OR, 0.38; 95%CI, 0.08-1.79) or drinking (adjusted OR, 0.36; 95%CI, 0.08-1.65) for risk of noncardia GC. Conclusions: Our data indicate no association between the two SNPs of IL-4-590 and IL-8-251 with overall GC risk, while the IL-8-251 TA or AA genotype conferred risk of cardia GC. Our findings contribute to the evidence body for risk of SNPs associated with the development of gastric cancer in this region.

• BMB Reports
• /
• v.35 no.6
• /
• pp.623-628
• /
• 2002

The Th1 vs. Th2 balance is critical for the maintenance of immune homeostasis. Therefore, the genes that are selectively-regulated by the Th1 and Th2 cytokines are likely to play an important role in the Th1 and Th2 immune responses. In order to search for and identify the novel target genes that are differentially regulated by the Th1/Th2 cytokines, the human PBMC mRNAs differentially expressed upon the stimulation with IL-4 or IL-12, were screened by employing the differential display-polymerase chain reaction. Among a number of clones selected, DC21 was identified as a novel target gene that is regulated by IL-4 and IL-12. The DC21 gene expression was up-regulated either by IL-4 or IL-12, yet counter-regulated by co-treatment with IL-4 and IL-12. DC21 is a dendritic cell protein with an unknown function. The sequence analysis and conserved-domain search revealed that it has two AU-rich motifs in the 3'UTR, which is a target site for the regulation of mRNA stability by cytokines, and that it belongs to the N-acetyltransferase family. The induction of DC21 by IL-12 peaked around 8-12 h, and lasted until 24 h. LY294002 and SB203580 significantly suppressed the IL-12-induced DC21 gene expression, which implies that PI3K and p38/JNK are involved in the IL-12 signal transduction pathway that leads to the DC21 expression. Furthermore, tissue blot data indicated that DC21 is highly expressed in tissues with specialized-resident macrophages, such as the lung, liver, kidney, and placenta. Together, these data suggest a possible role for DC21 in the differentiation and maturation of dendritic cells regulated by IL-4 and IL-12.

• The Journal of Korean Medicine
• /
• v.25 no.2
• /
• pp.207-219
• /
• 2004

Objectives : We investigated to find the relationship between single-nucleotide polymorphism (SNP) of IL4R, IL-10 and bee venom therapy efficacy in patients with RA treated with bee venom for 8 weeks. Methods : Korean RA patients (n=114) and healthy subjects (n=109) were included in this prospective study. Korean bee venom was dissolved in saline (diluted 1:3000) and administrated into acupuncture points. Bee venom therapy was applied twice a week and continued for 8 weeks. The clinical response was evaluated using various assessments before and after treatment. Disease severity was measured by determining the number of tender joints and swollen joints. Laboratory studies included ESR, CRP, and rheumatoid factor. Genotyping for IL-4R and IL-10 polymorphism was done by pyrosequencing analysis. Results : 1. In IL4R genotypes, there was significant difference between RA ptitients tind controls group. 2. In IL4R genotypes, there was significant difference among Good, Mild and Bad responders to in RA patients, but in the frequency of alleles and carriers, there were no significant difference. 3. There was no significant difference between RA patients and controls group in IL-10 gene genotypes. 4. In IL-10 genotypes, there was no significant difference among Good, Mild and Bad responders to in RA patients. 5. There was no significant difference in the improvement of ESR, CRP and KHAQ scores after bee venom therapy in RA patients among the IL4R or IL-10 genotypes. Conclusions : In IL-4R genotypes, there was significant difference between RA patients and control group, and among Good, Mild and Bad responders in RA patients. However, in IL-10 genotypes, there was no significant difference between RA patients and controls group and among Good, Mild and Bad responders in RA patients.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.20
• /
• pp.8775-8778
• /
• 2014

Background: To explore the relationship between polymorphisms of interleukin17 (IL-17) gene(A-832G 7488A/G) and the susceptibility to silicosis, a risk factor for lung cancer. Materials and Methods: A total of 113 silicosis patients and 116 workers without silicosis were enrolled in the case-control study. IL-17A A-832G and IL-17F 7488A/G polymorphisms were evaluated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequencies of AA,GG and AG of IL-17A A-832G locus in the case and control groups were 46.9%, 8.0%, 45.1%, and 49.2%, 7.6%, 43.2%, respectively, with no significant differences (p>0.05).The GG genotype in the IL-17F (7488A/G) locus was not found. The frequencies of AA and GA of IL-17F 7488A/G locus in the case and control groups were 84.1%, 15.9% and 66.4%, 33.6%, respectively (p<0.05). Analysis of combined effects showed that the individuals with GG+AG genotype of IL-17A and GG+GA genotype of IL-17F are protected against silicosis (OR=0.469). Conclusions: IL-17F 7488A/G is associated with susceptibility to silicosis, and G allele may have a protective effect. No relationship was found between IL-17A gene polymorphisms at A-832G and silicosis.

• Tuberculosis and Respiratory Diseases
• /
• v.41 no.5
• /
• pp.462-474
• /
• 1994

Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

• The Journal of Pediatrics of Korean Medicine
• /
• v.14 no.1
• /
• pp.79-116
• /
• 2000

The KGBT has been used to weak children with anorexia, fatigue, and growth retardation. This study was carried out to prove the effects of the hematopoiesis and the immune proliferation by KGBT. Previously, C57BL/6 mice was treated with cyclophosphamide(100mg/kg) for leukopenia, and then administered KGBT (concentration is 1.37 g/kg, 504 mg/kg, and 137 mg/kg) to the treated mice. The mice was analyzed expression of thrombopoietin(TPO), stem cell factor(SCF) and interleukin-3 from bone marrow cell, interleukin-10 (IL-10), and interferon-$ {\gamma}$(INF-${\gamma}$) from splenic cell, and NOSⅡ gene from macrophage using by RT-PCR. Also proliferation of immune cell was analyzed using 3H-thymidine uptake and flow cytometery in splenic cells. The results were obtained as follows ; 1. The total number of WBC, RBC and PLT was increased in the KGBT treated group than in the control group. 2. In vitro, the proliferation of splenic cells was increased in normal, control, and KGBT treated group. And Administration of KGBT was reduced the cytotoxicity by CTX. 3. In bone marrow cell, the gene expression of immune regulatory factor that associated with hematopoiesis, such as TPO, SCF, and IL-13 was increased in the KGBT treated group than control. 4 The titer of hemagglutinin and hemolysin was increased in the KGBT treated group than control. 5. In analysis of positive leucocytes from splenic cell of BALB/c mice, the subpopulation percent of CD4+, CD8+,and CD19+ was increased in the KGBT treated group than control. The KGBT has been used to weak children with anorexia, fatigue, and growth retardation. This study was carried out to prove the effects of the hematopoiesis and the immune proliferation by KGBT. Previously, C57BL/6 mice was treated with cyclophosphamide(100mg/kg) for leukopenia, and then administered KGBT (concentration is 1.37 g/kg, 504 mg/kg, and 137 mg/kg) to the treated mice. The mice was analyzed expression of thrombopoietin(TPO), stem cell factor(SCF) and interleukin-3 from bone marrow cell, interleukin-10 (IL-10), and interferon-$ {\gamma}$(INF-${\gamma}$) from splenic cell, and NOSⅡ gene from macrophage using by RT-PCR. Also proliferation of immune cell was analyzed using 3H-thymidine uptake and flow cytometery in splenic cells. The results were obtained as follows ; 1. The total number of WBC, RBC and PLT was increased in the KGBT treated group than in the control group. 2. In vitro, the proliferation of splenic cells was increased in normal, control, and KGBT treated group. And Administration of KGBT was reduced the cytotoxicity by CTX. 3. In bone marrow cell, the gene expression of immune regulatory factor that associated with hematopoiesis, such as TPO, SCF, and IL-13 was increased in the KGBT treated group than control. 4 The titer of hemagglutinin and hemolysin was increased in the KGBT treated group than control. 5. In analysis of positive leucocytes from splenic cell of BALB/c mice, the subpopulation percent of CD4+, CD8+,and CD19+ was increased in the KGBT treated group than control. 6. The expression of IL-10 gene was reduced in the KGBT treated group than control, whereas the expression of INF-${\gamma}$ was increased in the KGBT treated group. 7. In macrophage, the production of NO and gene expression of NOSH was increased in the KGBT treated group than control. 8. After infection of EMC virus, the survival time of infected mice was longer in the KGBT treated group than control.

• The Korea Journal of Herbology
• /
• v.23 no.2
• /
• pp.1-8
• /
• 2008

Objectives : The purpose of this study was to evaluate on the antimicrobial effect on the periodontal pathogens and anti-inflammatory effect of Artemisiae Iwayomogii Herba. Artemisiae Iwayomogii Herba has been used for treating as Artemisiae Capilaris Herba in Korea. Methods : Artemisiae Iwayomogii Herba was prepared by extracting medicinal herb with water. We investigated antimicrobial activity by the minimun inhibitory concentration (MIC) test. We also investigated inhibition of IL-$1{\beta}$-induced collagenase-l(MMP-l), stromelysin-1(MMP-3), interleukin-6 gene expression in human gingival fibroblasts. Results : The antimicrobial effect of Artemisiae Iwayomogii Herba was evaluated with MIC against periodontopathogens; Porphyromonas gingivalis 2561, W50, A7A1-28, 9-14K-1, Prevotella intermedia28, and Actinobacillus actinomycetemcomitans Y4, MICs of Artemisiae Iwayomogii Herba were 0.156 mg/ml, 0.625 mg/ml, 0.313 mg/ml, 1.25 mg/ml, 10 mg/ml and 10 mg/ml. The anti-inflammatory effect of Artemisiae Iwayomogii Herba was evaluated with Influence of herbs on the IL-$1{\beta}$-induced expression of MMP-1, MMP-3, interleukin-6, IL-$1{\beta}$ increased MMP-1, MMP-3, interleukin-6 mRNA levels. Artemisiae Iwayomogii Herba significantly inhibited IL-$1{\beta}$-induced MMP-1, MMP-3, interleukin-6 gene expressions in a dose-dependent manner. Conclusions : These results suggested that Artemisiae Iwayomogii Herba might reduce the excessive proteolytic capacity of the gingival fibroblast during inflammation and could be developed a new drug in periodontitis.