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Anti-oxidative and Anti-inflammatory Effect of Fractionated Extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (혈관내피세포에서 토복령(土茯苓)의 항산화 및 항염증 효과)

  • Lee, Chang-Hyun;Yi, Hyo-Seung;Kim, Jae-Eun;Heo, Sook-Kyoung;Cha, Chang-Min;Won, Chan-Wook;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.24 no.3
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    • pp.39-50
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    • 2009
  • Objectives : Smilacis glabrae rhizoma (SG) has been traditionally used as a herbal medication of musculoskeletal disorders like arthritis, pain, convulsions, and syphilis in traditional Korean medicine. This study was investigated anti-oxidative and anti-inflammatory effect of fractionated extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (HUVEC). Methods : SG extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of SG onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymethoxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of ICAM-1 and VCAM-1 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially ethyl acetate (EA) extract, significantly inhibited free radical generation, the TNF-$\alpha$-induced intracellular oxidation. Furthermore, the EA extract protected TNF-$\alpha$-induced adhesion to THP-1, expression of adhesion molecules accompanied by an attenuation of IL-6 and IL-8 formation in HUVEC. Conclusions : These results indicate that EA extract of SG have potential as an agent of atherosclerosis and other chronic inflammatory diseases including diabetes, hypertension, and arthritis.

Effects of Coptidis Rhizoma on the Anti-inflammation and Motor Recovery in Photothrombotic Brain Infarction Model in Rats (광화학적 뇌경색 백서 모델에서 황련의 항염증 및 운동기능 회복에 미치는 효과)

  • Lee, Su-Kyung;Lee, In;Shin, Sun-Ho;Kim, Eun-Young;Shin, Byung-Cheul
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.179-189
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    • 2009
  • Objectives : Coptidis Rhizoma (Coptis japonica MAKINO; CR) is a well known crude drug as antimicrobial, antibacterial, anti-inflammatory, antioxidant activity. However, there is no study of the effect of CR on brain infarction and it's mechanism. The aim of this study was to investigate the effects on ischemic stroke induced by photothrombotic infarction by evaluating the functional & neuronal recovery after brain infarction. Materials & Methods : Male Sprague-Dawley rats (250-300 g) were induced photothrombotic brain infarction on sensorimotor cortex, and brain infarction volume by image J software (NIH, USA) after Nissl stain, also single pellet reaching task as a functional motor recovery were observed. After orally pretreated by CR (500 mg/kg) or normal saline as a sham control before 7 days from the time of photothrombotic infarction, rats were sacrificed. After then we analysed anti-inflammatory cytokines (TNF-$\alpha$, IL-6, IL-1$\beta$), by RT-PCR and ELISA method, and immunohistochemistry (GFAP, connexin-43) as a marker of neural plasticity. Results : CR (100, 250, 500 mg/kg) decreased the infarction volume dose-dependently, however the effect of 500mg/kg of CR (CR 500) showed the best (P=0.051). Also, CR 500 decreased the infarction volume time-dependently, the most effective time was 3-7 days after stroke. Photothrombosis increased inflammatory cytokines after infarction, CR 500 suppressed significantly mRNA expression of IL-1$\beta$, IL-6 and TNF-$\alpha$. In serum, CR 500 decreased the amount of IL-1$\beta$, 12h, 24h and 48h respectively (p < 0.05), also decreased that of IL-6 and TNF-$\alpha$, 12h respectively (p < 0.05) after infarction. The more astrocytes were observed and neural plasticity was facilitated in the rat brain of CR 500 than that of sham control in immunohistochemistry. Conclusions : This results suggest that CR decrease infarction volume and improve functional motor recovery in acute stage in photothrombotic ischemic infarction model in the mechanism of anti-inflammation and promoting neural plasticity.

Suppressive Effects of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS) on TNCB(trinitrochlorobenzene)-induced Atopic Dermtitis NC/Nga Mice Model (TNCB로 유발한 아토피피부염 생쥐 모델에서 천일염가미방(天日鹽加味方)과 청기해독산(淸肌解毒散)의 병용 효과)

  • Lee, Kyoung-Mee;Gim, Seon-Bin;Choi, Hak-Ju;Choi, Jeong-June;Roh, Seong-Soo;Kim, Dong-Hee
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.215-224
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    • 2009
  • Objectives : Atopic dermatitis is a chronic inflammatory disease characterized by typically distributed eczematous skin lesion with pruritus, lichenification and dry skin. In this study, we performed to assess the therapeutic effects of co-treatment of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS, C&C) on the TNCB(trinitrochlorobenzene)-induced atopic dermatitis in NC/Nga mice, characterized by the onset of atopic dermatitis along with an increase the number of inflammatory cells and dysregulation of Th2 cytokines. Methods : Defined amount of CGB was sprayed on mice skin and CHS was simultaneously orally administrated to TNCB treated NC/Nga mice for 5 weeks. The immune cell types were caracterized by flow cytometry using each specific antibody. The amount of Th2 cytokines in serum and splenocytes culture supernatant were measured by ELISA. Results : Administration of C&C significantly reduced clinical dermatitis severity including pruritus, edema, eczematous and erythema. Histological findings indicated that the thickening of epidermis/dermis and dermal infiltration of inflammatory cells were dramatically reduced. Flow cytometry analysis showed that infiltrated immune cell numbers of CCR3+, B220+/IgE+, Gr-1+/CD11b+, and CD117+ were significantly reduced in C&C-treated dorsal skin lesion. Furthermore, T cell composition rate in PBMC was also dramatically decreased by the treatment. C&C greatly down-regulated production of Th2 cytokines including IL-4, IL-5, IL-13 in the serum. The down- regulatory effects of C&C on these Th2 cytokines production were also detected in CD3/ CD28 activated splenocytes. Conclusions : These results indicated that C&C is a plausible therapeutic agent for treatment of atopic dermatitis through regulating the Th2 skewed immune system.

Anti-inflammatory Effect of Eleutherococcus senticosus Extracts from Gangwon-do by Plant parts or Solvents (강원도 가시오갈피의 식물 부위 또는 추출 용매 조건에 따른 항염증 효과)

  • Junkyu Park;Mina Boo;Soojin An;Sujin Shin;Jinbong Park;Ho-Young Choi;Kyungjin Lee
    • The Korea Journal of Herbology
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    • v.38 no.4
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    • pp.21-29
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    • 2023
  • Background : The purpose of this study was to investigate the anti-inflammatory properties of stems and leaves of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. (ES) from Gangwon-do. Methods and Results : Stems and leaves of ES were collected from two areas in Gangwon-do: Cheorwon-gun and Samcheok-si. Samples were extracted with water by using the pressurized liquid extraction method and with 70% prethanol A by using the heat reflux extraction method. The anti-inflammatory effects of ES were evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT), lactate dehydrogenase(LDH) assay, nitric oxide(NO) assay, enzyme-linked immunosorbent assay(ELISA), and Western blot analysis in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). 1) Results showed that ES leaf extractions were not cytotoxic at a concentration of up to 30 ㎍/㎖. The leaves of 70% prethanol A extractions of ES(30 ㎍/㎖) inhibited NO, interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) production and decreased the protein level of cyclooxygenase 2(COX-2). There was no significant change in the protein level of inducible nitric oxide synthase(iNOS). The stem extractions of ES did not exhibit anti-inflammatory effects. Conclusions : In this study, the leaves of 70% prethanol A extractions of ES demonstrated anti-inflammatory effect on RAW 264.7 macrophages. The 70% prethanol A extractions have a relatively higher anti-inflammatory effect on RAW 264.7 macrophages than water extractions.

Resveratrol pretreatment alleviates NLRP3 inflammasome-mediated cardiomyocyte pyroptosis by targeting TLR4/MyD88/NF-κB signaling cascade in coronary microembolization-induced myocardial damage

  • Chang-Jun Luo;Tao Li;Hao-Liang Li;You Zhou;Lang Li
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.2
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    • pp.143-155
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    • 2023
  • Percutaneous coronary intervention and acute coronary syndrome are both closely tied to the frequently occurring complication of coronary microembolization (CME). Resveratrol (RES) has been shown to have a substantial cardioprotective influence in a variety of cardiac diseases, though its function and potential mechanistic involvement in CME are still unclear. The forty Sprague-Dawley rats were divided into four groups randomly: CME, CME + RES (25 mg/kg), CME + RES (50 mg/kg), and sham (10 rats per group). The CME model was developed. Echocardiography, levels of myocardial injury markers in the serum, and histopathology of the myocardium were used to assess the function of the cardiac muscle. For the detection of the signaling of TLR4/MyD88/NF-κB along with the expression of pyroptosis-related molecules, ELISA, qRT-PCR, immunofluorescence, and Western blotting were used, among other techniques. The findings revealed that myocardial injury and pyroptosis occurred in the myocardium following CME, with a decreased function of cardiac, increased levels of serum myocardial injury markers, increased area of microinfarct, as well as a rise in the expression levels of pyroptosis-related molecules. In addition to this, pretreatment with resveratrol reduced the severity of myocardial injury after CME by improving cardiac dysfunction, decreasing serum myocardial injury markers, decreasing microinfarct area, and decreasing cardiomyocyte pyroptosis, primarily by blocking the signaling of TLR4/MyD88/NF-κB and also reducing the NLRP3 inflammasome activation. Resveratrol may be able to alleviate CME-induced myocardial pyroptosis and cardiac dysfunction by impeding the activation of NLRP3 inflammasome and the signaling pathway of TLR4/MyD88/NF-κB.

Risk Assessment and Evaluation of Bt-transgenic Rice : Responses of Misgurnus anguillicaudatus and Cyprinus carpio fed on Bt-transgenic Rice Variety (해충저항성 Bt벼의 환경위해성 평가 : 해충저항성 Bt벼가 미꾸리(Misgurnus anguillicaudatus) 및 잉어(Cyprinus carpio)에 미치는 영향)

  • Oh, Sung-Dug;Lee, Dae-Yong;Sohn, Soo-In;Lee, Ki-Jong;Ryu, Tae-Hun;Lee, Jang-Yong;Park, Beom-Seok;Kweon, Soon-Jong;Suh, Seok-Cheol;Park, Jong-Sug
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.570-577
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    • 2011
  • We developed insect-resistant GM rice(Bt transgenic rice) by inserting the mCry1Ac1 a modified gene from the soil bacterium, Bacillus thuringiensis. The Bt transgenic rice expressing the Bttoxin mCry1Ac1 was tested for the effects on survival of Misgurnus anguillicaudatus and Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. M. anguillicaudatus and C. carpio fed 100% ground rice in suspension, using either Bt rice or non-GM counterpart rice(Nakdong). The Bt rice used for the test were confirmed to have the mCry1Ac1 gene expression by the immuno-strip and ELISA analysis. Feeding test showed that no significant differences in cumulative immobility and abnormal response of M. anguillicaudatus and C. carpio fed on between Bt rice and non-GM counterpart rice. The 96hr-LC50 values showed no difference between Bt rice(>1,000mg/L) and non-GM rice(>1,000mg/L). We concluded that there was no significant difference in toxicity for non-target organisms(M. anguillicaudatus and C. carpio) between Bt rice and non-GM counterparts.

Thymus quinquecostatus Extracts Suppress IFN-γ/TNF-α -induced Inflammatory Cytokine in Lung Fibroblast MRC-5 Cells

  • Young-Jae Song;Sa-Haeng Kang;Jae-Bin Seo;Se-Woong Ko;Ju-Ryoun Soh;Jeong-Hyang Park;Dong-Keun Kim;Tae-Hyun Kim;Hyuck-Se Kwon;Na Young Lee;Jong-Sik Jin
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.81-81
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    • 2020
  • Pneumonia is infection of air sacs in lungs by bacteria, viruses, fungi and abnormal atmosphere condition. That causes to inflammation and fills up with pus and fulid in air sacs. The symptoms of pneumonia are heavy cough, fever, shortness and rapid respiration and sputum with blood. Also, that remains sequela after the recovery. Thymus quinquecostatus (TQ) has a strong antibacterial effect. That has been used a natural medicine for bronchitis, asthma and nervous inflammation. In this study evaluated anti-inflammatory effects of TQ on IFN (interferon)-γ/TNF (tumor necrosis factor)-α (20 ng/ml) stimulation. MRC-5 cells were seeded at 1 × 105 cells/well in a 24-well plate and stabilized overnight at 37℃. The cells were treated with various concentrations of TQ extracts (DW, 30, 50, 70, 95% EtOH, 0.1~100 mg/ml) for 4h, Subsequently IFN-γ/TNF-α (20 ng/ml) was added to each well and incubated over 12h. the production of inflammatory cytokines were measured by Enzyme-linked immunosorbentassay(ELISA) method. The absorbance was measured at 450 nm using microplate reader. The treatment with TQ extract at dose of 0.1 to 100 mg/ml did not show cytotoxicity in MRC-5 cells (showed cytotoxicity at 95% EtOH 100 mg/ml). The results demonstrated that stimulation of TNF-α/IFN-γ significantly increased the production of inflammatory mediators in the MRC-5 cells compared with unstimulated cells. By contrast, treatment with TQ extreats doses of 0.001 to 10 mM significantly suppressed the production of IL-6 and IL-8. Especially. The higher percent of ethyl alcohol extracts suppressed the production of inflammatory cytokines. This results are considered that essential oil of TQ is extracted according to ethyl alcohol percent. Therefore, TQ has potential to mitigate pulmonary disease that can be used a medicine for pneumonia

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Antioxidant and anti-inflammatory effects and mechanism of Abeliophyllum distichum leaf extract in RAW264.7 macrophages (RAW264.7 대식세포에서 미선나무 잎 추출물의 항산화, 항염증 효능 및 기전연구)

  • Juhee Yoo;Kyung-Ah Kim
    • Journal of Nutrition and Health
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    • v.56 no.5
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    • pp.455-468
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    • 2023
  • Purpose: Abeliophyllum distichum (A.distichum) is a plant native to Korea. In this study, we investigated the mechanism of antioxidant and anti-inflammatory effects of the leaf extract of A.distichum. Methods: The antioxidant capacity of the A.distichum leaf extract was determined based on the total polyphenol content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay, and the ferric reducing antioxidant power (FRAP) assay. The anti-inflammatory effects of the A.distichum leaf extract were evaluated by measuring the production of nitric oxide (NO) and the expression levels of proinflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 using the enzyme-linked immunosorbent assay (ELISA) and reverse transcription quantitative real-time PCR (RT-qPCR). In addition, the expression of heme oxygenase-1 (HO-1), nuclear transcription factor-erythroid 2 related factor (Nrf2), inducible nitric oxide synthase (iNOS), and cyclooxygenase 2 (COX-2), as well as the activation of nuclear factorkappa B (NF-ĸB) were examined using the western blot analysis. Results: The total polyphenol content of the A.distichum leaf extract was 329.89 ± 30.17 gallic acid equivalents mg/g and the DPPH and ABTS scavenging activities were 55% and 70%, respectively. Additionally, the FRAP value of the extract was 743.68 ± 116.59 mg/mL. After 12-hour treatment with the A.distichum leaf extract, there was a tendency for the Nrf2 expression to increase, and the expression of HO-1 was significantly elevated in the RAW264.7 cells. The A.distichum leaf extract treatment resulted in decreased levels of NO, TNF-α, IL-6, and IL-1β, as well as reduced expression of iNOS and COX-2, along with inhibition of NF-κB activation in lipopolysaccharide-stimulated RAW264.7 cells. Conclusion: These results suggest that the A.distichum leaf extract exerts antioxidative and anti-inflammatory effects by upregulating the expression of HO-1 and downregulating NF-κB activation.

Cellular responses to 3D printed dental resins produced using a manufacturer recommended printer versus a third party printer

  • Beatriz Sona Cardoso;Mariana Brito da Cruz;Joana Faria Marques;Joao Carlos Roque;Joao Paulo Martins;Rodrigo Cordeiro Malheiro;Antonio Duarte da Mata
    • The Journal of Advanced Prosthodontics
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    • v.16 no.2
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    • pp.126-138
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    • 2024
  • PURPOSE. The aim of this study was to evaluate the influence of different 3D dental resins, using a manufacturer recommended printer and a third-party printer, on cellular responses of human gingival cells. MATERIALS AND METHODS. Three NextDent resins (Denture 3D+, C&B MFH and Crowntec) were used to produce specimens on printers NextDent 5100 (groups ND, NC and NT, respectively) and Phrozen Sonic Mini 4K (groups PD, PC and PT, respectively). Human gingival fibroblasts were cultured and biocompatibility was evaluated on days 1, 3 and 7. IL-6 and IL-8 concentrations were evaluated at 3 days using ELISA. Surface roughness was evaluated by a contact profilometer. SEM and fluorescence micrographs were analyzed at days 1 and 7. Statistical analyses were performed using SPSS and mean differences were tested using ANOVA and post-hoc Tukey tests (P < .05). RESULTS. There was an increase in cellular viability after 7 days in groups PC and PT, when compared to group PD. ND group resulted in higher concentration of IL-6 when compared to PT group. SEM and fluorescence micrographs showed less adhesion and thinner morphology of fibroblasts from group PD. No significant differences were found regarding surface roughness. CONCLUSION. The use of different printers or resins did not seem to influence surface roughness. NextDent 5100 and Phrozen Sonic Mini 4K produced resins with similar cellular responses in human gingival fibroblasts. However, Denture 3D+ resin resulted in significantly lower biocompatibility, when compared to C&B MFH and Crowntec resins. Further testing is required to support its long-term use, required for complete dentures.

Cellular Aging Inhibitory Effect of Perilla Leaf Extract on D-Galactose Induced C2C12 Myoblasts (D-갈락토스 유도 C2C12 근원세포에 대한 자소엽 추출물의 세포 노화 억제 효과)

  • Song-Mi Park;Sung-Woo Cho;Yung-Hyun Choi
    • Journal of Korean Medicine Rehabilitation
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    • v.34 no.2
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    • pp.15-28
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    • 2024
  • Objectives We used the D-galactose (D-gal) induced C2C12 myoblast senescence model to investigate whether ethanol extract of Perilla. fructescens leaves (EEPF) could delay cellular senescence and regulate related mechanisms. Methods C2C12 myogenic cells were cultured in an incubator under 37 ℃ and 5% CO2 conditions. EEPF, dried perilla leaves were pulverized and extracted at 1:10 (v/v) at 50 ℃ for 4 hours. Cell counting kit-8 and western blot analysis was performed. Annexin V-FITC apoptosis detection kit and DAPI staining was applied. Catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and malondialdehyde analysis kits were used. To measure the level of reactive oxygen species generation, staining and flow cytometry was used. To analyze the mitochondrial activity, membrane potential changes were measured using JC-1. 𝛽-gal activity was analyzed using SA-𝛽-gal staining solution, and DNA damage was analyzed by using 𝛾-H2AX. Quantikine ELISA kit was used to analyze inflammatory cytokine production. Results According to the results of this study, EEPF significantly alleviated the decrease in cell viability in C2C12 cells treated with D-gal and suppressed the decrease in the expression of proliferating cell nuclear antigen. EEPF also markedly blocked D-gal-induced C2C12 cell apoptosis and restored reduced activity of CAT, GSH-Px, T-AOC, SOD. In addition, EEPF suppressed the decrease in 𝛽-galactosidase activity, the induction of DNA damage and the increase in expression of senescence-associated secretory phenotype proteins such as p16, p53 and p21 in D-gal-treated C2C12 cells. Furthermore, EEPF significantly attenuated D-gal-induced production and expression of inflammatory cytokines such as interleukin (IL)-6 and IL-18. Conclusions The results of this study indicate that EEPF can be used as a potential candidate for the prevention and treatment of muscle aging.