• Asian Pacific Journal of Cancer Prevention
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• 제14권4호
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• pp.2295-2299
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• 2013

Background: To explore the role of caveolin-1(CAV-1) gene silencing on MAPK activation in lipopolysaccharide (LPS)-challenged human mammary epithelial cells. Methods: We established a MCF-10ACE of CAV-1 gene silencing from human mammary epithelial cell line MCF-10A by RNAi technology. DNA Microarray were used to detect the expression of inflammation-associated genes in MCF10ACE. Western blotting was used to examine the activation of MAPK in lipopolysaccharide(LPS)-challenged MCF-10A and MCF-10ACE. Moreover, immunofluorescence and Western bloting were performed to detect the co-localization of CAV-1 and toll-like receptor 4 (TLR4) in human mammary epithelial cells. Results: MCF-10ACE exhibited significant increases in inflammation-associated gene expression, especially IL-6 (~7-fold) and IL6R (~17-fold). In addition, LPS-induced p38 MAPK and JNK MAPK activation was significantly increased in MCF-10ACE. Furthermore, CAV-1 co-localized with TLR4 and appeared a negative correlation trend. Conclusion: CAV-1 gene silencing promotes MAPK activation via TLR4 signaling in human mammary epithelial cells response to LPS.

• Biomolecules & Therapeutics
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• 제23권1호
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• pp.84-89
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• 2015

We investigated the question of whether 7-oxygenated cholesterol derivatives could affect inflammatory and/or immune responses in atherosclerosis by examining their effects on expression of IL-23 in monocytic cells. $7{\alpha}$-Hydroxycholesterol ($7{\alpha}OHChol$) induced transcription of the TLR6 gene and elevated the level of cell surface TLR6 protein in THP-1 monocytic cells. Addition of an agonist of TLR6, FSL-1, to TLR6-expressing cells by treatment with $7{\alpha}OHChol$ resulted in enhanced production of IL-23 and transcription of genes encoding the IL-23 subunit ${\alpha}$ (p19) and the IL-12 subunit ${\beta}$ (p40). However, treatment with 7-ketocholesterol (7K) and $7{\beta}$-hydroxycholesterol ($7{\beta}OHChol$) did not affect TLR6 expression, and addition of FSL-1 to cells treated with either 7K or $7{\beta}OHChol$ did not influence transcription of the genes. Pharmacological inhibition of ERK, Akt, or PI3K resulted in attenuated transcription of TLR6 induced by $7{\alpha}OHChol$ as well as secretion of IL-23 enhanced by $7{\alpha}OHChol$ plus FSL-1. Inhibition of p38 MAPK or JNK resulted in attenuated secretion of IL-23. These results indicate that a certain type of 7-oxygenated cholesterol like $7{\alpha}OHChol$ can elicit TLR6-mediated expression of IL-23 by monocytic cells via PI3K/Akt and MAPKs pathways.

• 대한본초학회지
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• 제33권6호
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• pp.79-86
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• 2018

Objectives : Hemorrhoids are one of the most common diseases in humans. Jingyoganghwaltang (JG) and Cheongsimhwan (CS) have been used for treating hemorrhoids in Korean traditional clinical practice. The present study was designed to evaluate the traditional effects of JG and CS on the experimental hemorrhoid model in rats. Methods : Hemorrhoids are closely related to inflammation. Accordingly, we examined the nitric oxide (NO) production in macrophage cell line in order to evaluate the anti-inflammatory effect. The expression levels of inflammation related genes including IL-1 beta, IL-6, INOS, and TNF-alpha were examined via a real-time quantitative PCR. Croton oil-induced hemorrhagic animal model was used to test the in vivo efficacy against hemorrhoids. The rectal tissues were weighed and the inflammatory proteins were measured to confirm the anti-inflammatory effects. Results : JG and CS have a statistically significant effect on inhibition of NO production and on the reduction of inflammatory gene expression such as IL-1 beta, IL-6, INOS, and TNF-alpha. The synergistic effects of co-treatment of JG and CS were found out in the IL-6 gene expression. The in vivo study using croton oil-induced hemorrhoid model in rat was performed to check the co-treatment effects. As a result, the co-treatment reduced the inflammation of the rectal tissue and decrease the inflammation related protein productions including ICAM1, MMP2 and MMP9. Conclusions : These results suggest that JG and CS co-treatment demonstrated anti-inflammatory effects in croton oil-induced hemorrhoid model in rat.

• 한국동물학회지
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• 제39권1호
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• pp.115-121
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• 1996

임신중기에서 발기에 이르는 흰쥐의 태반에서 Placental Lactogen I (PL-I), II 그리고 Pit-1 의 유전자 발현 변화를 Northern blot hybridization으로 조사하였다. 그 결과, 임신시기에 따라 PL-I과 PL-II의 mRNA 양과 크기에 변화가 나타났다. 이들 유전자 발현에 미치는 에스트로겐의 영향을 조사하기 위하여, 임신 14일째 쥐의 난소를 제거하고(OVX), 이후 매일 에스트로겐을 투여한 후 (OVX+E), 임신 18일째 태반을 회수하여 PL-I, II 그리고 Pit-1의 유전자 발현을 Northern blot hybridization으로 조사하였다. OVX 군의 경우, PL-I의 mRNA 크기는 1 kb에서 1.3 kb로 PL-II의 mRNA는 0.6kb에서 1 kb로 변화하였다. OVX+E 군에서는 PL-I과 PL-II의 mRNA가 정상대조군과 같은 상태로 환원하였다. 정상대조군에 비하여 OVX와 OVX+E 군에서 PL-I과 PL-II의 mRNA 크기에는 영향을 미치지 못한 반면, mRNA 양은 난소제거시 감소하였다가, 에스트로겐을 투여하면, 부분적으로 회복되는 경향을 보였다. 이러한 본 실험의결과는 에스트로겐이 PL-I과 PL-II의 RNA splicing이나 polyadenylation 등을 포함한 유전자 발현에 영향을 미치며, Pit-1이 이 과정에 개입하는 것을 시사하는 것이다.

• Clinical and Experimental Pediatrics
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• 제48권2호
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• pp.197-203
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• 2005

목 적 : MMP 활동의 변화는 류마치스 관절염과 암 전위를 비롯한 여러 질환에서 보고되고 있다. 최근에 확장성 심근증에서 MMP 활동의 증가가 발표되었다. 아드리아마이신으로 유도된 심근증에서 MMP에 대한 보고는 없는 실정이다. 아드리아마이신으로 유도된 심근증에서 MMP, TIMP 유전자 발현을 연구하고 cytokine과의 관련성을 알아보고자 본 연구를 실시하였다. 방 법 : Sprague Dawley 쥐에 아드리아마이신 5 mg/kg을 1주일에 2번씩 2주간(누적 용량 : 20 mg/kg) 복강내 주사하였고, 정상쥐를 대조군으로 하였다. 2주 후에 쥐를 희생시켜서 혈청과 심장조직을 얻었다. 혈청에서 ELISA 원리로 MMP-2, TIMP-3, IL-6, TNF-${\alpha}$를 측정하였다. 심장에서 total RNA를 추출하였고, MMP-2, TIMP-3 IL-6, TNF-${\alpha}$ primer를 이용하여 PCR로 증폭하였다. 증폭된 DNA는 1% agarose gel에서 전기 영동하였고 UV light 아래에서 필름으로 촬영하였다. 결 과 : 혈청 MMP-2와 TIMP-3는 두 군 간에 유의한 차이가 없었다. 아드리아마이신군에서 IL-6은 $36.8{\pm}2.8pg/mL$, TNF-${\alpha}$은 $2.2{\pm}2.7pg/mL$로 정상군에 비해 유의한 증가를 보였다. 혈청 MMP-2와 TNF-${\alpha}$와는 r=0.41로 유의한 상관관계가 있었다. 심근 조직에서 MMP-2, IL-6, TNF-${\alpha}$는 발현되지 않았고, TIMP-3는 아드리아마이신군에서 대조군에 비해 유전자 발현이 감소되었다. 결 론 : 아드리아마이신으로 유도된 급성 심근증 모델에서는 심근에서 MMP, IL-6, TNF-${\alpha}$가 발현되지 않았고, TIMP 발현이 감소함을 알 수 있었다. 혈청 MMP와 TNF-${\alpha}$와의 상관성이 유의하게 높았으므로 TNF-${\alpha}$가 MMP 발현을 조절함을 시사해 준다. 앞으로 만성 심근증 모델에서 MMP, TIMP 발현에 대하여 연구할 예정이다.

• 동의생리병리학회지
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• 제17권6호
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• pp.1514-1518
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• 2003

Angelica sinensis radix, Danggui, is a traditional oriental medication, which has been used to modulate immune response. We report here that aqueous extract of Angelica sinensis radix (ASR) can induces NO production, and inhibit LPS-induced NO production in dose-dependent manner in RAW 264.7 macrophage cells. ASR also induces iNOS mRNA and iNOS protein expression, and exhibit inhibitory effect on iNOS mRNA and protein expression in a dose-dependent manner in LPS-stimulated RAW 264.7 macrophage cells. Cytokines involved in the regulation of inflammatory reaction and immune response may play a role in the pathogenesis. ASR induces. pro-inflammatory cytokine gene expression (IL-1α, IL-1β and IL-6 gene) in a dose-dependent manner, and inhibits the expressions of these cytokines in LPS-stimulated RAW 264.7 macrophage cells. These data indicate that (1) ASR may be a potential therapeutic modulator of NO synthesis in various pathological conditions, and (2) the immunomodulatory effects of ASR may be, in part, associated with the inducing or suppression of pro-inflammatory cytokine gene expressions.

• 대한의생명과학회지
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• 제24권2호
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• pp.94-101
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• 2018

Tuberculosis (TB) is infectious disease caused by Mycobacterium tuberculosis (MTB) infection. It is known that not only the property of microorganism but also the genetic susceptibility of infected patients is controlled. Interleukin 2 (IL-2) is a cytokine belonging to type 1 T helper (Th1) activity. In addition, IL-2, when infected with MTB, binds IL-2 receptor and promotes T cell replication and is involved in granuloma formation. The aim of this study was to investigate the genetic polymorphisms of the IL-2 receptor gene in tuberculosis patients and normal individuals. We analyzed 22 SNPs in three genes using the genotype data of 443 tuberculosis cases and 3,228 healthy controls from the Korea Association Resource for their correlation with tuberculosis case. IL2RA, IL2RB, and IL2RG genes were genotyped of 16, 4, and 2 SNPs, respectively. Among three genes, only IL2RA gene polymorphisms showed statistically significant association with tuberculosis case. 6 SNPs with high significance were identified in the IL2RA gene. In addition, the linkage disequilibrium (LD) structure of IL2RA gene was confirmed. SNP imputation of IL2RA gene was performed, it was confirmed that more SNPs were significant between case and control. If we look at the results of IL2RA gene analysis above, we can see that genetic polymorphism in the gene expressing $IL-2R{\alpha}$ will regulate the expression level of $IL-2R{\alpha}$, and the change in the immune system involved in $IL-2R{\alpha}$. In this study, genetic polymorphism that may affect host immunity suggests that susceptibility to tuberculosis may be controlled.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.837-845
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• 2014

Background: Overweight and obesity are recognized as major drivers of cancers including breast cancer. Several cytokines, including interleukin-6 (IL-6), IL-10 and lipocalin 2 (LCN2), as well as dysregulated cell cycle proteins are implicated in breast carcinogenesis. The nuclear, casein kinase and cyclin-dependent kinase substrate-1 (NUCKS-1), is a nuclear DNA-binding protein that has been implicated in several human cancers, including breast cancer. Objectives: The present study was conducted to evaluate NUCKS-1 mRNA expression in breast tissue from obese patients with and without breast cancer and lean controls. NUCKS-1 expression was correlated to cytokine profiles as prognostic and monitoring tools for breast cancer, providing a molecular basis for a causal link between obesity and risk. Materials and Methods: This study included 39 females with breast cancer (G III) that was furtherly subdivided into two subgroups according to cancer grading (G IIIa and G IIIb) and 10 control obese females (G II) in addition to 10 age-matched healthy lean controls (G I). NUCKS-1 expression was studied in breast tissue biopsies by means of real-time PCR (RT-PCR). Serum cytokine profiles were determined by immunoassay. Lipid profiles and glycemic status as well as anthropometric measures were also recorded for all participants. Results: IL-6, IL-12 and LCN2 were significantly higher in control obese and breast cancer group than their relevant lean controls (p<0.05), while NUCKS-1 mRNA expression was significantly higher in the breast cancer group compared to the other groups (p<0.05). Significant higher levels of IL-6, IL-12, and LCN2 as well as NUCKS-1 mRNA levels were reported in G IIIb than G IIIa, and positively correlated with obesity markers in all obese patients. Conclusions: Evaluation of cytokine levels as well as related gene expression may provide a new tool for understanding interactions for three axes of carcinogenesis, innate immunity, inflammation and cell cycling, and hope for new strategies of management.

• Journal of Animal Science and Technology
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• 제62권3호
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• pp.385-395
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• 2020

Polyinosinic:polycytidylic acid (poly[I:C]) can stimulate Toll-like receptor 3 (TLR3) signaling pathways. In this study, DF-1 cells were treated with poly(I:C) at various concentrations and time points to examine the comparative expression patterns of innate immune response genes. The viability of DF-1 cells decreased from 77.41% to 38.68% when cells were treated different dose of poly(I:C) from 0.1 ㎍/mL to 100 ㎍/mL for 24 h respectively. The expressions of TLR3, TLR4, TLR7, TLR15, TLR21, IL1B, and IL10 were increased in dose- and time-dependent manners by poly(I:C) treatment. On the contrary, the expression patterns of interferon regulatory factors 7 (IRF7), Jun proto-oncogene, AP-1 transcription factor subunit (JUN), Nuclear Factor Kappa B Subunit 1 (NF-κB1), and IL8L2 were varied; IRF7 and IL8L2 were increasingly expressed whereas the expressions of JUN and NF-κB1 were decreased in a dose-dependent manner after they were early induced. In time-dependent analysis, IRF7 expression was significantly upregulated from 3 h to 24 h, whereas JUN and NF-κB1 expressions settled down from 6 h to 24 h after poly(I:C) treatment although they were induced at early time from 1 h to 3 h. Poly(I:C) treatment rapidly increased the expression of IL8L2 from 3 h to 6 h with a plateau at 6 h and then the expression of IL8L2 was dramatically decreased until 24 h after poly(I:C) treatment although the expression level was still higher than the non-treated control. These results may provide the basis for understanding host response to viral infection and its mimicry system in chickens.

• 동의생리병리학회지
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• 제17권3호
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• pp.780-786
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• 2003

This study was carried out to know the immunity responses of the Spatholubus suberectus Dunn(hereinafter referred to STSD) to the synovial cells isolated from patients with rheumatoid arthritis. Various experiments were performed in vitro to analyse the immunity effects of STSD. Gene expression and production of pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, iNOS and COX-2 were determined by RT-PCR and ELISA kit. And also the binding activity of NF-kB and AP-1 were measured by Electromobility shift assay (EMSA) and the production of ROS was measured by flow cytometry. The results were obtained as follows 1. The gene expression and production of pro-inflammatory cytokines IL-1β, IL-6, TNF-α were reduced significantly. 2. The gene expression of iNOS and COX-2 were reduced. 3. The binding activity of NF-kB and AP-1 were inhibited. 4. The production of ROS in human synovial cells was reduced significantly. Comparison of the results for this study showed that STSD had immunomodulatory effects of suppressing or enhancing. So we expect that STSD should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.