• Title/Summary/Keyword: ex vivo study

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Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

  • Lee, Hyun-Kyoung;Jung, Jun-Yang;Lee, Sang-Hwa;Seo, Su-Yeong;Suh, Duk-Joon;Park, Hwan-Tae
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.3
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    • pp.161-168
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    • 2009
  • In the peripheral nerves, injury-induced cytokines and growth factors perform critical functions in the activation of both the MEK/ERK and JAK/STAT3 pathways. In this study, we determined that nerve injury-induced ERK activation was temporally correlated with STAT3 phosphorylation at the serine 727 residue. In cultured Schwann cells, we noted that ERK activation is required for the serine phosphorylation of STAT3 by neuropoietic cytokine interleukin-6 (IL-6). Serine phosphorylated STAT3 by IL-6 was transported into Schwann cell nuclei, thereby indicating that ERK may regulate the transcriptional activity of STAT3 via the induction of serine phosphorylation of STAT3. Neuregulin-1 (NRG) also induced the serine phosphorylation of STAT3 in an ERK-dependent fashion. In contrast with the IL-6 response, serine phosphorylated STAT3 induced by NRG was not detected in the nucleus, thus indicating the non-nuclear function of serine phosphorylated STAT3 in response to NRG. Finally, we determined that the inhibition of ERK prevented injury-induced serine phosphorylation of STAT3 in an ex-vivo explants culture of the sciatic nerves. Collectively, the results of this study show that ERK may be an upstream kinase for the serine phosphorylation of STAT3 induced by multiple stimuli in Schwann cells after peripheral nerve injury.

The Effect of Homnis Placenta Herbal Acupuncture on Reducing Expression of LPS-induced Arthritis Model as an Anti-inflammatory Agent (자하차약침(紫何車藥針)의 항염증능(亢炎症能)이 LPS 유발 관절염(關節炎) 치료(治療)에 미치는 영향(影響))

  • Park, Ki-Bum;Baek, Seung-Tae;Lee, Seung-Deok;Kim, Kyung-Ho;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.23 no.6
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    • pp.103-115
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    • 2006
  • Objectives : Rheumatoid arthritis(RA) is a systemic & a chronic inflammatory autoimmune disease . A chronic , locally destructive inflammmatory reaction in human is examplified by the synovitis present in some connective tissue disorder. The presence of a number of cytokines, $TNF-{\alpha}$, iNOS & expression of nitric oxide, NF-kB p65 activation implies an important role of cellular immune response in RA inflammatory reaction. This study was designed to evaluate on the effects of the Homnis Placenta herbal acupuncture on EX-LE201 & ST 35 reducing expression of LPS-induced arthritis model in mice. Materials and Methods : Homnis Placenta herbal acupuncture was inserted into 10 rats induced rheumatoid arthritis. The acupunctures were injected into the EX-LE201 and ST35 points. Such indexes were measured the inhibition of inducible nitric oxide synthase(iNOS) expression, nitric oxide(NO) production in vitro experiment and Tumor Necrosis $Factor-{\alpha}(TNF-{\alpha})$ & Nuclear Factor kappa $B(NF-{\kappa}B)$ p65 activation, synovial hyperplasia, angiogenesis and fibrosis in synovial membrane of knee joint of mice in vivo experiment. Results : 1.Homnis Placenta Herbal acupuncture inhibited iNOS mRNA and NO in RAW 264.7 cell of LPS-induced rheumatoid arthritis in a dose dependent manner. 2.Homnis Placenta Herbal acupuncture also showed significant inhibition of $TNF-{\alpha}$ & $NF-{\kappa}B$ p65, activation, synovial hyperplasia, angiogenesis and fibrosis in synovial membrane of knee joint of mice. Conclusion : These results suggest that Homnis Placenta Herbal acupuncture has an therapeutic effects on LPS induced-rheumatoid arthritis by inhibiting $TNF-{\alpha}$ activation.

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Antiplatelet and Antithrombotic Effects of the Extract of Lindera obtusiloba Leaves

  • Kim, Jun Ho;Lee, Jaemin;Kang, Soouk;Moon, Hongsik;Chung, Kyung Ho;Kim, Kyoung Rak
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.659-664
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    • 2016
  • Lindera obtusiloba has been used in traditional herbal medicine for the treatment of blood stasis and inflammation. The leaves of Lindera obtusiloba have been reported to exhibit various physiological activities. However, there is little information available on their antiplatelet and antithrombotic activities. Thus, the present study aimed to evaluate the effect of Lindera obtusiloba leaf extract (LLE) on platelet activities, coagulation and thromboembolism. In a platelet aggregation study, LLE significantly inhibited various agonist-induced platelet aggregations in vitro and ex vivo. Furthermore, LLE significantly inhibited collagen-induced thromboxane A2 (TXA2) production in rat platelets. In addition, oral administration of LLE was protective in a mouse model of pulmonary thromboembolism induced by intravenous injection of a mixture of collagen and epinephrine. Interestingly, LLE did not significantly alter prothrombin time (PT) and activated partial thromboplastin time (aPTT). This study indicates that the antithrombotic effects of LLE might be due to its antiplatelet activities rather than anticoagulation. Taken together, these results suggest that LLE may be a candidate preventive and therapeutic agent in cardiovascular diseases associated with platelet hyperactivity.

Effect of Codonopsis lanceolatae Extracts on Mouse IL-2, IFN-${\gamma}$, IL-10 Cytokine Production by Peritoneal Macrophage and the Ratio of IFN-${\gamma}$, IL-10 Cytokine (더덕 추출물의 경구 투여가 마우스의 사이토카인 생성과 IFN-${\gamma}$, IL-10 Ratio에 미치는 영향)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.22 no.1
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    • pp.69-74
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    • 2009
  • Codonopsis lanceolatae have been used as one of the traditional remedies as well as food source. We previously reported that in vitro supplementation of Codonopsis lanceolatae water extracts enhanced the splenocytes proliferation compared to the control group. This study, the combined immunomodulative effect of water extract Codonopsis lanceolatae was Seven to eight weeks old mice(balb/c) was fed ad libitum on chow diet and water extract of Codonopsis lanceolatae was orally administrated every other day for four weeks at two different concentrations(50 and 500 mg/kg B.W.). The production of cytokine(IL-2, IL-10 and IFN-${\gamma}$), secreted by macrophages stimulated with LPS or not, were detected by ELISA assay using the cytokine kit. The result of ex vivo study showed that the IL-2, IL-10 and IFN-${\gamma}$ was detected at 500 mg/kg B.W. supplementation group with LPS stimulation in all cases. Also, ratio of IFN-${\gamma}$, IL-10 was the range of 3${\sim}$7 with mitogen stimulation such as Con A and LPS. In conclusion, this study suggests that Codonopsis lanceolatae extracts may enhance the immune function by regulating the cytokine(IL-2, IL-10 and IFN-${\gamma}$) prodution capacity by activated macrophages in mice.

Effect of Corn Extracts on Mouse IL-2 Cytokine Production by Peritoneal Macrophage and the Ratio of IFN-${\gamma}$, IL-10 Cytokine (옥수수 추출물의 경구 투여가 사이토카인 IL-2 생성과 IFN-${\gamma}$와 IL-10 Ratio에 미치는 영향)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.25 no.2
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    • pp.362-367
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    • 2012
  • Corn has been used for a long time as a traditional remedy, as well as a food source. We previously reported that in vitro supplementation of corn water extracts enhanced the proliferation of splenocytes, compared to the control group. In this study, we examined the immunomodulative effect of a water extract of corn. Seven to eight weeks old mice(Balb/c) were fed an ad libitum chow diet, and were orally administrated a water extract of corn every other day, for four weeks, at two different concentrations(50 and 500 mg/kg B.W). Cytokine production(IL-2, IL-10 and IFN-${\gamma}$) by macrophages stimulated with LPS or not stimulated with LPS was detected by ELISA assay using the cytokine kit. In an ex vivo study, the cytokines IL-2, IL-10 and IFN-${\gamma}$ were detected at 500 mg/kg b.w. supplementation group with LPS stimulation in all cases. Also, the ratio of IFN-${\gamma}$ to IL-10 was in the range of 0~3 with mitogen stimulation, such as con A and LPS. In conclusion, this study suggests that in mice, corn extracts may enhance immune function by regulating the cytokine production(IL-2, IL-10 and IFN-${\gamma}$) of the activated macrophages.

Efficacy of Caffeine in Promoting Hair Growth by Enhancing Intracellular Activity of Hair Follicles

  • Kim, Sehyun;Kim, Su Na;Jeong, Gyusang;Hong, Min Jung;Lee, Yonghee;Shin, Seung Hyun;Park, Hyeokgon;Jung, Yu Chul;Kim, Eun Joo;Park, Byung Cheol;Kim, Hyoung-June
    • Korea Journal of Cosmetic Science
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    • v.1 no.1
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    • pp.11-18
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    • 2019
  • Caffeine is widely used in cosmetics and hair care products. Although its efficacy in stimulating hair growth has been confirmed in recent studies, its mechanism of action remains unelucidated. The present study aimed to determine the effects of caffeine on hair growth, with a focus on intracellular hair follicle activity. Experiments included in vitro and ex vivo tests, and a clinical study. Caffeine enhanced the cellular activity and potassium channel opening. It also promoted human hair follicle elongation. Immunohistochemical staining showed that the Ki-67 signal was significantly higher in cells treated with caffeine. These efficacies of caffeine were comprehensively demonstrated in clinical results, wherein caffeine-containing shampoo improved hair density after 24 weeks of testing. Collectively, the results of this study demonstrated that caffeine promoted hair growth and inhibited the progression of hair loss by enhancing intracellular activity of hair follicles.

Hot Water Extract of Scutellaria baicalensis Inhibits Migration, Invasion and Tube Formation in a Human Umbilical Vein Endothelial Cell Model and a Rat Aortic Ring Sprouting Model (혈관내피세포와 흰쥐 대동맥 미세혈관 발아 모델을 이용한 황금 열수추출물의 세포의 이동, 침투 및 관형성 억제 연구)

  • Kim, Eok-Cheon;Bae, Kiho;Kim, Han Sung;Yoo, Yeong-Min;Gelinsky, Michael;Kim, Tack-Joong
    • Journal of Life Science
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    • v.26 no.1
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    • pp.91-100
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    • 2016
  • Angiogenesis is essential for the pathophysiological processes of embryogenesis, tissue growth, diabetic retinopathy, psoriasis, wound healing, rheumatoid arthritis, cardiovascular diseases, and tumor growth. Inhibition of angiogenesis represents an attractive therapeutic approach for the treatment of angiogenic diseases such as cancer. However, uncontrolled angiogenesis is also necessary for tumor development and metastasis. Inhibition of vascular endothelial growth factor (VEGF) signaling, a critical factor in the induction of angiogenesis, cause robust and rapid changes in blood vessels of tumors and therefore VEGF constitutes a target for such anti-angiogenic therapy. Recently, since natural compounds pose significantly less risk of deleterious side effects than synthetic compounds, a great many natural resources have been assessed for useful substance for anti-angiogenic treatment. Here we evaluated the anti-angiogenic effects of a hot water extract of Scutellaria baicalensis (SBHWE) using in vitro assays and ex vivo animal experiments. Our results show that SBHWE dose-dependently abrogated vascular endothelial responses by inhibiting VEGF-stimulated migration and invasion as well as tube formation in a human umbilical vein endothelial cell (HUVEC) model, without cytotoxicity, as determined by a cell viability assay. Further study revealed that SBHWE prevented VEGF-induced neo-vascularization in a rat aortic ring sprouting model. Taken together, our findings reveal an anti-angiogenic activity of Scutellaria baicalensis and suggest that SBHWE is a novel candidate inhibitor of VEGF-induced angiogenesis.

Ex vivo Morphometric Analysis of Coronary Stent using Micro-Computed Tomography (미세단층촬영기법을 이용한 관상동맥 스텐트의 동물 모델 분석)

  • Bae, In-Ho;Koh, Jeong-Tae;Lim, Kyung-Seob;Park, Dae-Sung;Kim, Jong-Min;Jeong, Myung-Ho
    • Journal of the Korean Society of Radiology
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    • v.6 no.2
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    • pp.93-98
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    • 2012
  • Micro-computed tomography (microCT) is an important tool for preclinical vascular imaging, with micron-level resolution. This non-destructive means of imaging allows for rapid collection of 2D and 3D reconstructions to visualize specimens prior to destructive analysis such as pathological analysis. Objectives. The aim of this study was to suggest a method for ex vivo, postmortem examination of stented arterial segments with microCT. And ex vivo evaluation of stents such as bare metal or drug eluting stents on in-stent restenosis (ISR) in rabbit model was performed. The bare metal stent (BMS) and drug eluting stent (DES, paclitaxel) were implanted in the left or right iliac arteries alternatively in eight New Zealand white rabbits. After 4 weeks of post-implantation, the part of iliac arteries surrounding the stent were removed carefully and processed for microCT. Prior to microCT analysis, a contrast medium was loaded to lumen of stents. All samples were subjected to an X-ray source operating at 50 kV and 200 ${\mu}A$ by using a 3D isotropic resolution. The region of interest was traced and measured by CTAN analytical software. Objects being exposed to radiation had different Hounsfield unit each other with values of approximately 1.2 at stent area, 0.12 ~ 0.17 at a contrast medium and 0 ~ 0.06 at outer area of stent. Based on above, further analyses were performed. As a result, the difference of lengths and volumes between expanded stents, which may relate to injury score in pathological analysis, was not different significantly. Moreover, ISR area of BMS was 1.6 times higher than that of DES, indicating that paclitaxel has inhibitory effect on cell proliferation and prevent infiltration of restenosis into lumen of stent. And ISR area of BMS was higher ($1.52{\pm}0.48mm^2$) than that of DES ($0.94{\pm}0.42mm^2$), indicating that paclitaxel has inhibitory effect on cell proliferation and prevent infiltration of restenosis into lumen of stent. Though it was not statistically significant, it showed that the extent of neointema of mid-region of stents was relatively higher than that of anterior and posterior region in parts of BMS as showing cross-sectional 2-D image. suggest that microCT can be utilized as an accessorial tool for pathological analysis.

Effect of Tuberculin Skin Test on Ex-vivo Interferon-gamma Assay for Latent Tuberculosis Infection (투베르쿨린 검사가 결핵에 대한 체외 IFN-γ 검사 결과에 미치는 영향)

  • Lee, Jung Yeon;Choi, Hee Jin;Cho, Sang-Nae;Park, I-Nae;Oh, Yeon-Mok;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.4
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    • pp.406-412
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    • 2005
  • Background : Recently, two commercialized whole-blood assays, $QuantiFERON^{(R)}-TB$ Gold (QFT) and T $SPOT-TB^{(R)}$ (SPOT), which measure the $IFN-{\gamma}$ released in the whole blood after being incubation with mycobacterial antigens, were approved for the diagnosis of a latent tuberculosis infection (LTBI). However, there is data on whether or not the previously used PPD skin tests (TST) have any influence on the diagnostic ability of these ex-vivo $IFN-{\gamma}$ assays. Methods : Forty-six 15 year-old students who did not appear to be infected with Mycobacterium tuberculosis were enrolled in this study. The peripheral blood was collected and used for two $IFN-{\gamma}$ assays. The $IFN-{\gamma}$ assays and TST were performed at the baseline ($1^{st}$). The TST was repeated two months later ($2^{nd}$), and the $IFN-{\gamma}$ assays were repeated two ($2^{nd}$) and four months ($3^{rd}$) later only in those subjects who had negative results at the baseline in both the $IFN-{\gamma}$ assays and TST. An induration size > 10 mm was considered to be positive in the TST. Results : The mean TST value was $3.1{\pm}5.4mm$ (range: 0-20). Of the 46 subjects examined, 13 subjects (28.3%) showed positive results in the two-step TST. Nine (19.6%) were SPOT-positive and only one (2.2%) was QFT-positive. The $2^{nd}$ and $3^{rd}$ QFT were carried out in 23 and 25 all-negative subjects, respectively, and all showed negative results. The $2^{nd}$ SPOT was performed in 23 subjects and only one (4.3%) showed a weak-positive result. Conclusion : Even though there were some discrepancies in the results of the two ex-vivo $IFN-{\gamma}$ assays, it appears that their results were not influenced by a previous TST carried out in two or four months earlier.

Effect of onion peel extract supplementation on the lipid profile and antioxidative status of healthy young women: a randomized, placebo-controlled, double-blind, crossover trial

  • Kim, Jungmi;Cha, Yong-Jun;Lee, Kyung-Hea;Park, Eunju
    • Nutrition Research and Practice
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    • v.7 no.5
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    • pp.373-379
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    • 2013
  • The consumption of fruits and vegetables that have high polyphenol content has been previously associated with a reduced risk for cardiovascular disease. We investigated the effects of onion peel extract on plasma total antioxidant capacity, lipid peroxidation, and leukocyte DNA damage. This study was a randomized, double-blind, placebo-controlled, crossover trial. Healthy female subjects received either onion peel extract or placebo (dextrin) for two weeks, underwent a 1-week washout period, and then received the other treatment for an additional two weeks. After two weeks of onion peel extract supplementation, the total cholesterol level, low-density lipoprotein cholesterol level, and atherogenic index significantly decreased (P < 0.05). No changes were observed in activities of erythrocyte antioxidant enzymes or levels of lipid peroxidation markers following onion peel extract supplementation. Additionally, no significant difference was found in plasma antioxidant vitamin (retinol, tocopherols, carotenoids, and coenzyme Q10) levels or ex vivo $H_2O_2$-provoked oxidative DNA damage after onion peel extract supplementation. The present interventional study provides evidence of the health benefits of onion peel extract and demonstrates its effects in modulating lipid profiles in healthy young Korean women.