Evaluation of ‘TNF-α, IL-6, and MMP-9’ Test Kit for Screening of Meibomian Dysfunction in Patients with Inflammatory Dry Eye Syndrome

Min-Hye Park,Jung-Eun Park,Jang-Won Byun,Min-Ji Choi,Il-Hoon Cho,Myeong-Jin Jeong,Yoon-Jung Choy,Koon-Ja Lee 대한시과학회 2020 대한시과학회지 Vol.22 No.1

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. Purpose : To evaluated the ‘TNF-α, IL-6, MMP-9’ test kit for screening of inflammatory dry eye and IDE (inflammatory dry eye) with MGD (meibomian gland dysfunction). Methods : A total of 118 dry eyes were selected using OSDI (ocular surface disease index) questionnaire among participated 20~30s without ophthalmologic diseases except for dry eye. Small amount of tear obtained from meniscus of the conjunctiva were tested with TNF-α, IL-6, and MMP-9 kit. IDE refers to the criteria which specifies the corneal staining and conjunctival hyperemia more than grade 1 and MGD refers to the criteria which specifies meibomian gland blockage and meibomian orifice obstruction with more than grade 1. Chi-square test was performed to analyze the correlation between the IDE, MGD and the results of ‘TNF-α, IL-6, MMP-9’ tests. and ROC (receiver operate characteristics) curve was used for the sensitivity, specificity and AUC (area under the curve) for the accuracy of ‘TNF-α, IL-6, MMP-9’ tests. Results : TNF-α and IL-6 were significantly correlated with IDE (p<.050) and ‘TNF-α, IL-6, MMP-9’ test kit showed a high agreement of 80.20% with MMP-9 test kit(p<.050) although the accuracy was lower than MMP-9 test kit. The MMP-9 showed no correlation with MGD, however TNF-α, IL-6 were significantly correlated with MGD (p<.050). sensitivity, specificity, and AUC of TNF-α, IL-6 tests for MGD were 85.50%, 34.70%, 0.601, 85.50%, 32.70%, and 0.591. The sensitivity, specificity, and AUC of ‘TNF-α, IL-6, MMP-9’ test kit for IDE with MGD were 100.00%, 34.10%, and 0.670, respectively, which shows higher accuracy than MMP-9. Conclusion : TNF-α and IL-6 tests are useful for the diagnosis of MGD, and ‘TNF-α, IL-6, MMP-9’ test kit is useful for screening IDE with MGD.

Anti-inflammatory Effects of Ethanolic Extracts from Codium fragile on LPS-Stimulated RAW 264.7 Macrophages via Nuclear Factor kappaB Inactivation

Yoon, Ho-Dong,Jeong, Eun-Ji,Choi, Ji-Woong,Lee, Min-Sup,Park, Myoung-Ae,Yoon, Na-Young,Kim, Yeon-Kye,Cho, Deuk-Moon,Kim, Jae-Il,Kim, Hyeung-Rak The Korean Society of Fisheries and Aquatic Scienc 2011 Fisheries and Aquatic Sciences Vol.14 No.4

Bacterial lipopolysaccharide (LPS) induces expression of pro-inflammatory cytokines and enzymes producing nitric oxide (NO) and prostaglandins (PGs) in immune cells. This process is mediated by the activation of nuclear factor kappaB (NF-${\kappa}B$). In this study, we investigated the anti-inflammatory characteristics of Codium fragile ethanolic extract (CFE) mediated by the regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using LPS-stimulated murine macrophage RAW 264.7 cells. CFE significantly inhibited LPS-induced NO and $PGE_2$ production in a dose-dependent manner and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells with no cytotoxicity. Pro-inflammatory cytokines, such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor-${\alpha}$, were significantly reduced by treatment of CFE in LPS-stimulated RAW 264.7 cells. CFE inhibited the promoter activity of (NF)-${\kappa}B$ in LPS-stimulated macrophages. Treatment with CFE suppressed translocation of the NF-${\kappa}B$ p65 subunit by preventing proteolytic degradation of inhibitor of ${\kappa}B-{\alpha}$. These results indicate that the CFE-mediated inhibition of NO and $PGE_2$ production in LPS-stimulated RAW 264.7 cells is mediated through the NF-${\kappa}B$-dependent transcriptional downregulation of iNOS and COX-2, suggesting the potential of CFE as a nutraceutical with anti-inflammatory activity.

Antioxidant Activity of Yogurt Fermented at Low Temperature and Its Anti-inflammatory Effect on DSS-induced Colitis in Mice

Ji-Woo Yoon,Sung-Il Ahn,Jin-Woo Jhoo,Gur-Yoo Kim 한국축산식품학회 2019 한국축산식품학회지 Vol.39 No.1

This study was performed to evaluate the antioxidant activity of yogurt fermented at low temperature and the anti-inflammatory effect it has on induced colitis with 2.5% dextran sodium sulfate (DSS) in Balb/c mice. Yogurt premix were fermented with a commercial starter culture containing Lactobacillus acidophilus, Bifidobacterium lactis, Streptococcus thermophilus, and Lactobacillus delbrueckii subsp. bulgaricus at different temperatures: 22℃ (low fermentation temperature) for 27 h and 37℃ (general fermentation temperature) for 12 h. To measure antioxidant activity of yogurt samples, DPPH, ABTS+ and ferric reducing antioxidant potential (FRAP) assays were conducted. For animal experiments, inflammation was induced with 2.5% DSS in Balb/c mice. Yogurt fermented at low temperature showed higher antioxidant activity than that of the yogurt fermented at general temperature. In the inflammatory study, IL-6 (interleukin 6) was decreased and IL-4 and IL-10 increased significantly in DSS group with yogurt fermented at general temperature (DYG) and that with yogurt fermented at low temperature (DYL) compared to that in DSS-induced colitic mice (DC), especially DYL had higher concentration of cytokines IL-4, and IL-10 than DYG. MPO (myeloperoxidase) tended to decrease more in treatments with yogurt than DC. Additionally, yogurt fermented at low temperature had anti-inflammatory activity, although there was no significant difference with general temperature-fermented yogurt (p>0.05).

Original Articles : Anti-inflammatory Effects of Ethanolic Extracts from Codium fragile on LPS-Stimulated RAW 264.7 Macrophages via Nuclear Factor kappaB Inactivation

( Ho Dong Yoon ),( Eun Ji Jeong ),( Ji Woong Choi ),( Min Sup Lee ),( Myoung Ae Park ),( Na Young Yoon ),( Yeon Kye Kim ),( Deuk Moon Cho ),( Jae Il Kim ),( Hyeung Rak Kim ) 한국수산과학회(구 한국수산학회) 2011 Fisheries and Aquatic Sciences Vol.14 No.4

Bacterial lipopolysaccharide (LPS) induces expression of pro-inflammatory cytokines and enzymes producing nitric oxide (NO) and prostaglandins (PGs) in immune cells. This process is mediated by the activation of nuclear factor kappaB (NF-κB). In this study, we investigated the anti-inflammatory characteristics of Codium fragile ethanolic extract (CFE) mediated by the regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using LPS-stimulated murine macrophage RAW 264.7 cells. CFE significantly inhibited LPS-induced NO and PGE2 production in a dose-dependent manner and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells with no cytotoxicity. Pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor-α, were significantly reduced by treatment of CFE in LPS-stimulated RAW 264.7 cells. CFE inhibited the promoter activity of (NF)-κB in LPS-stimulated macrophages. Treatment with CFE suppressed translocation of the NF-κB p65 subunit by preventing proteolytic degradation of inhibitor of κB-α. These results indicate that the CFE-mediated inhibition of NO and PGE2 production in LPS-stimulated RAW 264.7 cells is mediated through the NF-κB-dependent transcriptional downregulation of iNOS and COX-2, suggesting the potential of CFE as a nutraceutical with anti-inflammatory activity.

창출도인탕가미방(蒼朮桃仁湯加味方)이 류마토이드 관절염에 미치는 실험적 연구

박은숙 ( Eun Sook Park ),윤일지 ( Il Ji Yoon ) 한방재활의학과학회 2009 한방재활의학과학회지 Vol.19 No.4

Objectives: This study was carried out to understand the immune responses of the Changchuldointanggami-bang(after referred to CDIT) on antioxidants, THP-1 cell and rheumatoid arthritis in Collagen-induced Arthritis(CIA) mice. Methods: The antioxidative effect of CDIT on DPPH free radical and SOD-like activity was measured. CDIT was administered to THP-1 cell, cytokine and mRNA associated with inflammation were measured. CDIT was orallyadministered to mice with arthritis by collagen Ⅱ and then cytokine, PBMC in the serum were measured. Results: 1. The scavenging activity of CDIT on DPPH free radical was dose-dependent. 2. The effect of CDIT on SOD-like activity was dose-dependent. 3. The production of IL-1β, IL-6, TNF-α was decreased significantly in THP-1 cell. 4. The expression of IL-1β mRNA, IL-6mRNA, TNF-α mRNA were decreased significantly in THP-1 cell. 5. IL-1β, IL-6, TNF-α were decreased significantly in the serum of CIA mice. 6. CD3+, CD4+, CD8+ were increased significantly, butCD3+/CD69+, CD3+/CD49b+, B220+/CD23+ were decreased significantly in PBMC of CIA mice. Conclusions: Taking all these observations into account, CDIT is considered to be effective in rheumatoid arthritis. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.

다탐색(多探索) 법을 통한 저자극성 액체 세정제 조성물 개발

김배환 ( Peter Kim ),현기안 ( Ki-an Hyeon ),정지윤 ( Ji-youn Chung ),윤삼숙 ( Sanm-sook Yoon ),강한철 ( Han Chyul Kang ),박선희 ( Sun Hee Park ),고강일 ( Kang Il Ko ),김기호 ( Ki Ho Kim ) 대한화장품학회 2005 대한화장품학회지 Vol.31 No.1

세정제에 널리 상용화되어 있는 알킬에톡시설페이트 계의 음이온 계면활성제는 피부 흡착의 특성 때문에 충분히 헹구어 내지 않으면 피부에 잔존되어 자극의 원인이 되어 염증 반응을 일으키게 된다. 따라서 기존의 음이온 계면활성제를 대체 또는 보완하기 위해 기존의 계면활성제들을 단백질 변성 실험, 세포 독성 그리고 IL-1α 측정을 통해 선별하였다. 본 연구는 저자극성의 음이온 또는 비이온, 양쪽성 이온의 14종의 계면활성제와 민감성 피부를 위해 제조된 13종의 기존 세정제 제품에 대한 세포독성을 실시하여, 가장 독성이 낮은 계면활성제로 sodium laureth sulfate (음이온), sodium cocoyl isethionate (음이온), sodium lauroamphoacetate (양성이온), cocamidopropyl betaine (양성이온), alkyl polyglycoside (비이온)가 선택되었고 2종의 기존 세정제 제품을 비교 제형으로 선택하였다. 5종의 계면활성제를 20종의 formulation으로 제조하여 단백질 변성(<3M SLS (13.2%)), 세포독성 실험 및 폐첩포 실험을 통하여 다시 5종을 선별하였다. 이들 제형을 진피 배양으로 세포독성 및 IL-1α 방춘량을 조사하여 가장 자극이 낮은 계면활성제 제형을 선택하였으며, 첨가된 계면활성제의 자극을 완화하기 위하여 항염과 보습 효과가 우수한 마치현 추출물(3%, 5%)과 fructan (3%, 5%)을 농도별로 첨가한 제형을 제조하여 가장 안전성이 뛰어난 농도를 선택하였다. 최종적으로 선택된 제형 5번을 3차원 세포 배양을 통한 인공피부를 이용하여 가장 자극이 낮게 나타난 기존의 제품들과 세포 독성 및 IL-1α 방출량을 비교 조사하여 저자극성의 액체 세정제를 개발하였다. Alkyl ethoxy sulfate type surfactants, widely used in commercial cleansers, are easily adsorbed to skin to often cause skin irritation and inflammation if not thoroughly rinsed nut. In order to replace or complement existing surfactants, we screened the existing surfactants through protein denaturation method, cell cytotoxicity assay and human IL-1α assay, etc. Fourteen surfactants have been chosen from among too irritant anionic, cationic and/or zwitter-ionic ones and investigated for cell cytotoxicity in human fibroblast cell lines using monolayer culture with the thirteen commercially available cleansers for sensitive skin. From these results, we selected 5 surfactants and 2 commercial cleansers (names not shown), such as sodium laureth sulfate (anionic), sodium cocoyl isethionate (anionic), sodium lauroamphoacetate (zwitter-ionic), and cocamidopropyl betaine (zwitter-ionic), alkyl polyglycoside (non-ionic). 20 formulations were made out of 5 surfactants and five of them were chosen through a protein denaturation method (lower than 3 M sodium dodecyl sulfate solution (13.2%)), cell cytotoxicity and human patch test. These five selected formulations containing preservatives were compared to two selected commercial cleansers by cell cytotoxicity and human IL-1α ELISA assay using dermal equivalent. Finally, we selected the best formulation. To this formulation, fructan (3% or 5%) or/and portulaca extract (3% or 5%) well known for its anti-inflammatory and moisturizing effects were added and investigated for cell cytotoxicity using dermal equivalent. In cytotoxicity assay using dermal equivalent, two formulations containing 5% fructan and 3% or 5% portulaca extract were less toxic than the others. In cytotoxicity assay and human IL-1α ELISA using 3D culture, the selected formulation containing 5% fructan and 5% portulaca extract showed better efficiency than those of the others and 2 commercial cleansers. As a result, we could develop a low irritant and safe liquid cleanser.

관절1호방이 Collagen Ⅱ로 유발된 관절염 억제에 관한 연구

한규진 ( Kyu Jin Han ),진광선 ( Kwang Seon Jin ),신혜란 ( Hye Ran Shin ),윤일지 ( Il Ji Yoon ),최승훈 ( Seung Hoon Choi ),오민석 ( Min Seok Oh ) 한방재활의학과학회 2005 한방재활의학과학회지 Vol.15 No.4

Objectives : The aim of this study was to know the immunity responses of Gwanjulbang-1(Quanjiefang-1)(hereinafter referred to GJB-1) to on Rheumatoid Arthritis in Collagen-induced Arthritis(CIA) Mice Methods : For this purpose, experiments were performed to measure the cytotoxicity against mLFC, hFLSs and the production of proinflammatory cytokines IL-1β, IL-6, TNF-α, IFN-γ in hFLSs and the production of pro-inflammatory cytokines IL-4, IL-10 and the value of CD3+(T), CD19+(B), CD3+/CD69+, CD4, γδ T cells, CD4+/CD25+ Results : 1. The cytotoxicity against mLFC, hFLSs were not measured. 2. The production of pro-inflammatory cytokines IFN-γ were reduced in hFLSs. 3. The production of pro-inflammatory cytokines IL-4, IL-10 were increased. 4. Statistically significant value of CD3+(T), CD19+(B), CD3+/CD69+, CD4, γδ T cells, CD4+/CD25+ compared with control group, wild type, MTX. Conclusions : Comparison of the results for this study showed that GJB-1 had immunomodulatory effects of suppressing or enhancing. So we expect that GJB-1 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.

Indomethacin으로 유발된 흰쥐의 위장장애에 ChondroT가 미치는 영향

김주일 ( Joo-il Kim ),김선길 ( Sun-gil Kim ),김지훈 ( Ji-hoon Kim ),윤찬석 ( Chan-suk Yoon ),최지민 ( Ji-min Choi ),최찬헌 ( Chan-hun Choi ),김선종 ( Seon-jong Kim ) 한방재활의학과학회 2020 한방재활의학과학회지 Vol.30 No.3

Objectives The aim of this study was to investigate the inhibitory effect of ChondroT in indomethacin-induced gastric mucosal injury rat model. Methods Sprague-Dawley rats were randomly assigned to intact, control Joins, Celebrex, ChondroT50 and ChondroT200. Indomethacin (25 mg/kg) was used to induce damage to the gastric mucosal injury. ChondroT was administered by orally to inhibit the indomethacin-induced gastric mucosal injury. At the end of the experiment, pH level in stomach, stomach contents volume, tumor necrosis factor-α (TNF- α) level, interleukin-1β (IL-1β) level, prostaglandin E2 (PGE2) level, myeloperoxidase (MPO) activity, erythrocytes, and thrombocytes were measured. Ophthalmologic and histopathological examination was also analyzed. Results pH level in stomach and Stomach contents volume had no difference between Control, PC-Joins, PC-Cele, ChondroT50 and ChondroT200 group. TNF-α level was decreased in PC-Joins, PC-Cele, ChondroT50 and ChondroT200 group and there were no significant difference. IL-1β level was decreased in PC-Joins group and ChondroT200 group compared to control group. PGE2 level had no significant difference between Control, PC-Joins, PC-Cele, ChondroT50 and ChondroT200 group. MPO level and complete blood count level were decreased in PC-Joins, PC-Cele, ChondroT50 and ChondroT200. Symptom score of ophthalmologic examination was decreased in ChondroT50 and ChondroT200 group compared to control group. Conclusion Based on these results, It could be suggested that ChondroT was effective in reducing damage to the gastric mucosal injury. And further study is needed to conduct a rigorous clinical research. (J Korean Med Rehabil 2020;30(3):57-69)

Original Articles : Ethanolic Extract of Chondria crassicaulis Inhibits the Expression of Inducible Nitric Oxide Synthase and Cyclooxygenase-2 in LPS-Stimulated RAW 264.7 Macrophages

( Yeon Kye Kim ),( Eun Ji Jeong ),( Min Sup Lee ),( Na Young Yoon ),( Ho Dong Yoon ),( Jae Il Kim ),( Hyeung Rak Kim ) 한국수산과학회(구 한국수산학회) 2011 Fisheries and Aquatic Sciences Vol.14 No.4

Inflammatory mediators such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) have been implicated in various inflammatory diseases. In this study, we investigated the anti-inflammatory activities of Chondria crassicaulis ethanolic extract (CCE) by measuring its effects on the expression of iNOS and COX-2 proteins in lipopolysaccharide (LPS)-treated RAW 264.7 murine macrophages. CCE significantly and dose-dependently inhibited the LPS-induced release of nitric oxide and prostaglandin E2, and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells, without causing any cytotoxicity. It also inhibited the production of the pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in LPS-stimulated RAW 264.7 cells. Moreover, treatment with CCE strongly suppressed nuclear factor-κB (NF-κB) promoter-driven expression in LPS-treated RAW 264.7 cells. CCE treatment blocked nuclear translocation of the p65 subunit of NF-κB by preventing proteolytic degradation of inhibitor of κB-α. These results indicate that CCE regulates iNOS and COX-2 expression through NF-κB-dependent transcriptional control, and identifies potential candidates for the treatment or prevention of inflammatory diseases.

Reversal of serologic, immunologic, and histologic dysfunction in mice with systemic lupus erythematosus by long‐term serial adipose tissue–derived mesenchymal stem cell transplantation

Choi, Eun Wha,Shin, Il Seob,Park, So Young,Park, Ji Hyun,Kim, Jong Sung,Yoon, Eun Ji,Kang, Sung Keun,Ra, Jeong Chan,Hong, Sung Hwa Wiley Subscription Services, Inc., A Wiley Company 2012 Vol.64 No.1

<P><B>Abstract</B></P><P><B>Objective</B></P><P>To investigate the efficacy of human adipose tissue–derived mesenchymal stem cell (AD‐MSC) transplantation in systemic lupus erythematosus (SLE) and to determine the optimal transplantation window for stem cells either before or after disease onset.</P><P><B>Methods</B></P><P>(NZB × NZW)F<SUB>1</SUB> mice with SLE were administered human AD‐MSCs (5 × 10<SUP>5</SUP>) intravenously every 2 weeks from age 6 weeks until age 60 weeks, while the control group received saline vehicle on the same schedule. Another experiment was carried out with a different initiation time point for serial transplantation (age 6 weeks or age 32 weeks).</P><P><B>Results</B></P><P>Long‐term serial administration (total of 28 times) of human AD‐MSCs ameliorated SLE without any adverse effects. Compared with the control group, the human AD‐MSC–treated group had a significantly higher survival rate with improvement of histologic and serologic abnormalities and immunologic function, and also had a decreased incidence of proteinuria. Anti–double‐stranded DNA antibodies and blood urea nitrogen levels decreased significantly with transplantation of human AD‐MSCs, and serum levels of granulocyte–macrophage colony‐stimulating factor, interleukin‐4 (IL‐4), and IL‐10 increased significantly. A significant increase in the proportion of CD4+FoxP3+ cells and a marked restoration of capacity for cytokine production were observed in spleens from the human AD‐MSC–treated group. In the second experiment, an early stage treatment group showed better results (higher survival rates and lower incidence of proteinuria) than an advanced stage treatment group.</P><P><B>Conclusion</B></P><P>Serial human AD‐MSC transplantation had beneficial effects in the treatment of SLE, without adverse effects. Transplantation of human AD‐MSCs before disease onset was preferable for amelioration of SLE and restoration of immune homeostasis.</P>