Inflammation is a protective mechanism against pathogens, but if maintained continuously, it destroys tissue structures. Aggregatibacter actinomycetemcomitans is a gram-negative, facultative anaerobic bacterium often found in severe periodontitis. A. actinomycetemcomitans invades epithelial cells and triggers inflammatory response in the immune cells. In this study, we investigated the effect of water-soluble rosehip extract on A. actinomycetemcomitansinduced inflammatory responses. A human monocytic cell line (THP-1) was differentiated to macrophages by phorbol 12-mystristate 13-acetate treatment. The cytotoxic effect of extract was determined using the 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide assay. The effects of extract on bacterial growth were examined by measuring the optical densities using a spectrophotometer. THP-1-derived macrophages were infected A. actinomycetemcomitans after extract treatment, and culture supernatants were analyzed for cytokine production using enzyme-linked immunosorbent assay. Protein expression was measured by western blotting. Extract was not toxic to THP-1- derived macrophages. A. actinomycetemcomitans growth was inhibited by 1% extract. The extract suppressed A. actinomycetemcomitans-induced tumor necrosis factor-α, interleukin (IL)-1β, and IL-8 production. It also decreased mitogen-activated protein kinase (MAP kinase) and nuclear factor-κB (NF-κB) phosphorylation. Moreover, the extract inhibited the expression of inflammasome components, including nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3, Absent in Melanoma 2, and apoptosis associated speck-like protein containing a CARD. And cysteine-aspartic proteases-1 and IL-1β expression were decreased by the extract. In summary, extract suppressed A. actinomycetemcomitans growth and decreased inflammatory cytokine production by inhibiting activation of MAP kinase, NF-κB, and inflammasome signaling. Rosehip extract could be effective in the treatment of periodontal inflammation induced by A. actinomycetemcomitans infection.

목적 : 마이봄샘기능저하증(meibomian gland dysfunction, MGD)을 수반하는 염증성 건성안의 감별진단에 대한 ‘TNF-α, IL-6, MMP-9’ 검사키트의 유용성을 평가하였다. 방법 : 건성안 이외의 안질환이 없는 20~30대 중 OSDI 설문 검사에 따른 건성안 총 118안을 대상하였고, 결막낭 메니스커스로부터 소량의 눈물을 채취하여 TNF-α, IL-6 및 MMP-9 검사를 하였다. 각막염색과 결막충혈 이 모두 Grade 1 이상인 경우는 염증성 건성안으로, 마이봄샘폐쇄와 마이봄샘구멍막힘이 모두 grade 1 이상인 경우는 MGD 관련 건성안으로 평가하였다. 염증성 건성안 및 MGD와 TNF-α, IL-6, MMP-9과의 상관성은 카 이제곱검정(Chi-square test)으로 분석하였고, ‘TNF-α, IL-6, MMP-9’ 검사키트의 염증성 건성안과 MGD를 수반하는 염증성 건성안 감별능력은 ROC 커브를 이용하여 민감도, 특이도 및 AUC(Area under the curve)를 구하고 정확도를 평가하였다. 결과 : 염증성 건성안은 TNF-α와 IL-6와 유의한 상관성을 보였고(p＜.050), ‘TNF-α, IL-6, MMP-9’ 검사 키트는 MMP-9 검사키트와 80.20%의 높은 일치도를 나타냈으나(p＜.050), 염증성 건성안 감별에 대한 민감도, 특이도, 정확도는 MMP-9 검사키트보다 낮았다. MGD는 MMP-9 검사와 상관성을 보이지 않았고, TNF-α와 IL-6 검사와는 유의한 상관성을 보였으며, MGD 감별에 대한 민감도, 특이도, 정확도는 각각 85.50%, 34.70%, 0.601, 85.50%, 32.70%, 0.591로 나타났다. MGD 수반한 염증성 건성안 감별에 대한 ‘TNF-α, IL-6, MMP-9’ 검사키트의 민감도, 특이도 및 정확도는 100.00%, 34.10%, 0.670로 MMP-9 검사키트보다 더 높았다. 결론 : MGD 진단에는 TNF-α, IL-6 검사가 유용하며, ‘TNF-α, IL-6, MMP-9’ 검사키트는 MGD를 수반한 염증성 건성안 평가에 유용할 것으로 사료된다.

Previous studies have investigated the potential relationship between promoter polymorphism (-308, G/ A) of tumor necrosis factor (TNF)-α and various autoimmune diseases. However, results from published data were inconclusive. To verify relationship between TNF-α polymorphism (-308, G/A) and susceptibility to autoimmune diseases such as vitiligo, celiac disease, and rheumatoid arthritis, we have performed a metaanalysis with all relevant articles before October 2016. The electronic search of PubMed, google, and Embase databases was performed to identify eligible studies investigating the relationship of TNF-α polymorphism with autoimmune diseases including vitiligo, celiac disease, and rheumatoid arthritis. Genotype frequency data of TNF-α polymorphism (-308, G/A) were extracted and the meta analysis was performed by Comprehensive meta-analysis program with odds ratio (OR) and 95% confidence intervals (95% CI). Genotype models were applied with dominant and recessive models and allele model analyzed. The final analysis included 37 publication papers with a total of 6,102 autoimmune disease patients and 6,987 control subjects. In result, a statistical significant correlation between TNF-α polymorphism (-308, G/A) and susceptibility to autoimmune disease was not detected in our meta-analysis (p>0.05 in all models). Our results suggest that the TNF-α polymorphism might not be related to the development of autoimmune disease. If further results in larger studies would be accumulated in the future, this relationship would be clarified.

Mushroom is known for anti-inflammatory and anti-oxidative potential. This study provides evidence that theinhibitory effect of mushroom on the expression of pro-inflammatory cytokines in human keratinocytes, HaCaT cells. To definethe underlying mechanisms of action, tumor necrosis factorα/IFNγ-activated human keratinocytes model was used. Mushroomsignificantly inhibited the expression of cytokines in HaCaT cells. Taken together, the results demonstrate that mushroom inhibitedinflammtion, suggesting that mushroom (DW extract: Grifola frondosa Cordyceps militaris), (Ethanol extract: Ganoderma lucidum,Lentinus edodes, Cordyceps militaris, Flammulina velutipes) might be a candidate for the treatment of skin inflammation.

Aggregatibacter actinomycetemcomitans is an important pathogen in the development of localized aggressive periodontitis. Lipopolysaccharide (LPS) is a virulent factor of periodontal pathogens that contributes to alveolar bone loss and connective tissue degradation in periodontal disease. Our present study was designed to investigate the cytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine gene expression profiling in RAW 264.7 cells was performed by microarray analyses. The cytokine mRNA and protein levels and related signaling pathways induced by Aa LPS were measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly induced the expression of NF-κB, NF-κB-related genes, inflammatory cytokines, TNF-α and IL-1β in RAW 264.7 cells. NF-κB inhibitor pretreatment significantly reduced the levels of TNF-α and IL-1β mRNA and protein. In addition, the Aa LPS-induced TNF-α and IL-1β expression was inhibited by p38/JNK MAP kinase inhibitor pretreatment. These results show that Aa LPS stimulates TNF-α and IL-1β expression through NF-κB and p38/JNK activation in RAW 264.7 cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.

생체 내 실험에서 발효 인삼꽃 추출물(FM), 발효하지 않은 인삼꽃 추출물(FD)과 대조군으로 생리식염수를 2주간 마우스 체중 ㎏ 당 100, 200 ㎎/㎏ B.W.의 농도로 마우스에 경구 투여한 후 LPS에 의해 활성화된 복강 대식세포가 분비하는 염증성 사이토카인 IL-6, TNF-α의 생성량을 측정하였다. 그 결과, IL-6는 발효 LPS로 자극한 경우, 두 가지 농도에서 모두 처리한 군에 비해 높은 증식능을 나타내었고, LPS로 자극한 결과, 특히 발효 인삼꽃 추출물 200 ㎎/㎏ B.W. 농도에서 유의적으로 낮은 IL-6 분비량을 보였다. TNF-α의 경우, 100 ㎎/㎏ B.W.와 200 ㎎/㎏ B.W. 두 농도 모두에서 LPS로 자극하지 않은 경우, 낮은 증식 효과를 보여주었고, 자극한 경우, 인삼꽃 시료를 첨가한 군이 대조군보다 낮은 TNF-α 분비량을 보였으며, FM에서 FD보다 더 TNF-α를 억제하는 효과가 큰 것을 볼 수 있었다. 이상의 결과에 따르면 FD의 사이토카인 IL-6, TNF-α 생성 효과는 200 ㎎/㎏ B.W. 농도 투여 시 효과적으로 면역 세포와 면역 기관의 주요 기능을 증진시킬 가능성이 있을 것으로 사료된다. 이러한 연구결과를 토대로 앞으로 인삼꽃 발효를 이용한 기능성 사료 개발과 더불어 산업적 측면에서 보다 긍정적인 효과를 얻을 수 있을 것으로 판단된다.

Tumor necrosis factor alpha (TNFα) is a multifunctional inflammatory cytokine that regulates various cellular and bio-logical processes. Increased levels of TNFα have been im-plicated in a number of human diseases including diabetes and arthritis. Sympathetic nervous system stimulation via the beta2-adrenergic receptor (β2AR) in osteoblasts suppresses osteogenic activity. We previously reported that TNFα up- regulates β2AR expression in murine osteoblastic cells and that this modulation is associated with TNFα inhibition of osteoblast differentiation. In our present study, we explored whether TNFα induces β2AR expression in human osteo-blasts and then identified the downstream signaling path-way. Our results indicated that β2AR expression was increa-sed in Saos-2 and C2C12 cells by TNFα treatment, and that this increase was blocked by the inhibition of NF-κB acti-vation. Chromatin immunoprecipitation and luciferase reporter assay results indicated that NF-κB directly binds to its cog-nate elements on the β2AR promoter and thereby stimulates β2AR expression. These findings suggest that the activation of TNFα signaling in osteoblastic cells leads to an upregu-lation of β2AR and also that TNFα induces β2AR exp-ression in an NF-κB-dependent manner.

Tumor necrosis factor alpha (TNFα) is a multifunctional cytokine that is elevated in inflammatory diseases such as atherosclerosis, diabetes and rheumatoid arthritis. Recent evidence has suggested that β2 adrenergic receptor(β2AR) activation in osteoblasts suppresses osteogenic activity. In the present study, we explored whether TNFα modulates βAR expression in osteoblastic cells and whether this regulation is associated with the inhibition of osteoblast differentiation by TNFα. In the experiments, we used C2C12 cells, MC3T3- E1 cells and primary cultured mouse bone marrow stromal cells. Among the three subtypes of βAR, β2 and β3AR were found in our analysis to be upregulated by TNFα. Moreover, isoproterenol-induced cAMP production was observed to be significantly enhanced in TNFα-primed C2C12 cells, indicating that TNFα enhances β2AR signaling in osteoblasts. TNFα was further found in C2C12 cells to suppress bone morphogenetic protein 2-induced alkaline phosphatase (ALP) activity and the expression of osteogenic marker genes including Runx2, ALP and osteocalcin. Propranolol, a β2AR antagonist, attenuated this TNFα suppression of osteogenic differentiation. TNFα increased the expression of receptor activator of NF-κB ligand (RANKL), an essential osteoclastogenic factor, in C2C12 cells which was again blocked by propranolol. In summary, our data show that TNFα increases β2AR expression in osteoblasts and that a blockade of β2AR attenuates the suppression of osteogenic differentiation and stimulation of RANKL expression by TNFα. These findings imply that a crosstalk between TNFα and β2AR signaling pathways might occur in osteoblasts to modulate their function.

Chronic inflammatory diseases such as Crohn′s disease and ulcerative colitis are associated with increased risk of colon adenocarcinoma. Apoptic induction of colon cancer cells by cytokines and death receptors is an important anti-cancer therapy. We observed that co-administration of TNFα and IFNγ in human colon cancer cell line, HCT116, resulted in cell death and expression of IL-32. Cleavage forms of caspase-3, caspase-9, and PARP were increased in TNFα / IFNγ-treated HCT116. mRNA expression of death receptors, including TNFR1 and Fas were not changed and NO generation was not induced by combination of TNFα and IFNγ. However, mRNA expression of IL-32α, β, and γ was increased in TNFα / IFNγ-treated HCT116. To determine the effect of IL-32 in HCT116 cell apoptosis by TNFα / IFNγ stimulation, IL-32 siRNA-transfected HCT116 cells were cultured with TNFα / IFNγ and cell proliferation was measured. IL-32 siRNA induced slight recovery of cell viability of TNFα / IFNγ-stimulated HCT116. These results suggest that IL-32 is not directly related to apoptosis of HCT116 by TNFα / IFNγ stimulation. However, IL-32 expression by TNFα or TNFα / IFNγ in a colon cancer cell line is very interesting because of the unknown effect of IL-32 in colon cancer. Our study will contribute to development of studies for IL-32 function in human colon cancer and anti-cancer therapies using cytokines.