• Title/Summary/Keyword: Antiinflammatory

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Analysis of Chlorogenic Acid Content and Biological Activities of Aralia elata Ethanol Extract (두릅 에탄올 추출물의 Chlorogenic acid 함량 분석 및 생리활성)

  • Lee, Jeong Ho;Jeong, Kyoung Ok;Im, So Yeon;Jin, Da Mon;Lee, Wang Ro
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.574-585
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    • 2022
  • This study was conducted to quantify chlorogenic acid content and evaluate biological activity, such as antioxidant, antibacterial, anti-inflammatory, and digestive enzyme activity of Aralia elata ethanol extract (AEE). The SC50 of DPPH and ABTS radical scavenging activities of AEE were 4.79±0.05 mg/mL, 5.79±0.05 mg/mL; total polyphenol and total flavonoid contents were 170.0±1.8 mgGAE/g, 105.5±4.1 mgQE/g, respectively. Nitric oxide (NO) was increased in RAW 264.7 cells and Caco-2 cells with treatment of LPS, and production of NO was inhibited by AEE in a concentration-dependent manner. Production of NO was reduced by 60.0±1.1% in RAW 264.7 cells and 50.7±2.8% in Caco-2 cells at of AEE. Similarly, the production of inflammatory cytokines (TNF-α, IL-1β and IL-6) was inhibited in a concentration dependent manner. Antibacterial activity increased as the dose concentration of AEE increased, and the MIC was 75 mg/mL for L. monocytogenes, and 100 mg/mL for S. typhimurium and H. pylori. In addition, amylase and protease enzyme activity was observed in AEE and increased enzyme activity was observed according to the concentration of the extract. AEE contained 7.06±0.01 mg/g of chlorogenic acid. As a result of the experiment, it is judged that it can be used as basic data for the development of health food using Aralia elata.

Anti-inflammatory effect of naringenin-7-O-phosphate in LPS-induced RAW 264.7 cells (LPS로 유도된 RAW 264. 7 대식세포에서 Naringenin-7-O-phosphate의 항염증 활성)

  • Hyehyun Hong;Tae-Jin Park;Byeong Min Choi;Yu-Jung Yi;Seung-Young Kim
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.46-52
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    • 2023
  • The most abundant flavanone of grapefruits, naringenin (NN), is well known for its hepatoprotective, anti-lipid peroxidation and anti-carcinogenic effects. We generated three derivatives from NN using this technique in previous studies. Among them, it was confirmed that naringenin-7-O-phosphate (N7P), whose biological and physicochemical properties were not reported, showed a water solubility 45 times higher than that of NN. Therefore, in this study, the anti-inflammatory activity was evaluated in RAW 264.7 cells to investigate the potential physiological activity of N7P. As a result, N7P showed nitric oxide (NO) inhibitory activity at concentrations that did not show toxicity. In addition, prostaglandin E2 (PGE2) showed significant inhibitory activity from the lowest concentration of 12.5 μM and showed increased inhibitory activity compared to NN. In addition, as a result of western blot, N7P showed increased cyclooxygenase-2 (COX-2) inhibitory activity than NN, and effectively inhibited NO and PGE2 by significantly inhibiting their expression pathways. N7P also inhibited inflammatory cytokines, including tumor necrosis factor-α, interleukin-6. Based on these results, we propose that N7P can be used as a potent antiinflammatory agent.

Study on the Herb Remedies of ENT, Eyes, Teeth and Skin Problems (이비인후, 안, 치아 및 피부증상의 민간요법에 관한 고찰)

  • Cho, Kyoul-Ja;Kang, Hyun-Sook
    • Journal of East-West Nursing Research
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    • v.1 no.1
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    • pp.50-71
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    • 1997
  • The intention of this study is to apprehend the con. tents and methods of herb remedies that are commonly conducted when there are health-problem cases of ENT, eyes, teeth and skin. Methods of this study are divided into two stages : 1) For a period of six months from December 1994 to June 1995, some 40 persons who are believed to be well versed with herb remedies have been randomly chosen, and we made a survey on herb remedies by symptoms: and 2) we have endeavored to make their grounds evident through the studies on literatures with the focus on the basic data collected. Their results are as follows: 1) When one feels a pain in ears, such herb remedies are employed as pouring the vapor into ears, which is made by steaming Alaska pollack, or as applying or wiping with the juices of radish or the ginkgo, or' alum. Applying the radish juice is effective for sterilization and fever removal: and applying the ginkgo juice is effective for cleanliness. But, plastering alum, sesame oil or castor oil, or pouring the vapor of Alaska pollack into ears are perhaps effective but do not have any pharmacologic grounds. 2) When one bleeds at the nose, such kinds of herb remedies are applied as stimulating nose or head with cold water, pressing nose or ridge of nose, or filling up nares with mashed mugworts. In addition, they have utilized garlic or leeks. Such methods as stimulating with cold water or just pressing nose and ridge of nose is based on reasonable grounds, i.e. vasoconstriction and vascular compression ; and applying mashed garlic on the sole of foot is good for the circulation of Qui ; and the use of mugworts and leeks is based upon the pharmacological function of hemostasis. 3) When one feels a sore throat such kinds of herb remedies are employed as gargling or rinsing throat with brine, drinking hot gruel or water, or drinking the juice of mugwort, radish, ginger or Chinese quince. Gargling with brine or drinking the juice of mugwort, radish or ginger is based upon the pharmacological function of pain alleviation, fever removal, and detoxication. 4) When a boil is formed in mouth, such herb remedies are applied as spreading honey, brine or alum water, and taking gall nut, Chinese matrimony vine, lotus root, etc, for drugs. Spreading honey, brine or water that is made by infusing gallnut, Chinese matrimony vine, lotus root is based upon such functions as hematosis, astriction, antibacterial, and antiphlogistic, Alum, eggplant and licorice are said to be effective, but their pharmacological effects have no grounds. 5) When one has conjunctivitis such herb remedies are commonly applied as irrigation with brine and dropping breast milk in eyes. Moreover, such other drugs are used as plantain. shepherd's purse, and purslane, etc. The use of brine, breast milk, plantain, shepherd's purse and purslane is based upon such functions as sterilization, antiphlogistic, disinfection and pain relieving. Eriocaulon sieboldianum, bean stem, bean pod and narcissus leaves are said to be effective, but their pharmacological action have no basis. When one has a stye, such herb remedies are applied as extracting eyelashes, stimulating by a massage of middle finger, third finger or big toe, as well as sear ing with a heated bamboo comb that is fine-toothed. Other than these, plantain and nightshade's nuts are used as drugs for it. Extracting eyelashes corresponds with exclusing suppurative node and draining the stye of pus ; and the use of plantain is based upon disinfection: and nightshade's nuts are said to be effective, however, their pharmacological action has no grounds. 6) For a treatment of toothache, such herb remedies are commonly employed as rinsing mouth with brine and holding cold water or gasoline in the mouth ; and as the drugs that are believed to be effective have been Welsh onion, ginger and castor-oil, plant, etc. The use of Welsh onion is based upon pain killing, antiinflammatory actions, and the use of ginger is based upon detoxication and disinfection ; and seeds of castor-oil plants are said to be effective, but they have no pharmacological basis. 7) When one has hives, such herb remedies are commonly applied as rubbing burned straw in affected parts, exposing to its smoke, rubbing with salt, sweeping down with a broom, and spreading and drinking boiled water of trifoliate orange. The use of cassia tora seeds, walnut, aloe and radish is said to be effective. The use of cassia tor a seeds has the functions of intestinal order, anti-paralysis, etc. The use of walnut has resulted in an increase of blood by invigorating spirits ; and the use of aloe is based upon disinfection, antibiotic, anti-salt, antihistamine and detoxication action. But, the effects of radish juice and straw's smoke have no pharmacological grounds. 8) When one gets a boil, such herb remedies are commonly used as applying a plaster, paste of flour mixed with yolk, soy sauce or honey, as well as spreading pounded elm tree. Other remedies that have been said to be effective are ; heating with mugwort, brine, wild rocambole, aloe, onion, squid's bone, etc. The use of mugwort is based upon pain killing, astringent antiinflammatory and tranquility. Wild rocambole is based upon the generation and maintenance functions of cell-joining textures ; elm tree upon antiphlogistic ; aloe upon fever removal and antiphlogistic ; onion on pain killing, fever removal, antiphlogistic and tranquility ; squid's bone on astriction: and brine or vinegar on sterilization. Pine resin and gardenia seed are said to be effective, but they have no pharmacological basis. 9) When one cuts his skin, such herb remedies are commonly employed as spreading mugwort's juice or squid's bone powder, or pressing the wounds. In addition, kalopanax, onion and fine soil are employed. The use of mugwort, kalopanax and squid's bone is based upon such functions as hemostasis, sedation, pain killing, antibacterial ; and fine soil is said to be effective, but it has no pharmacological basis. 10) When one suffers from whitlow, such herb remedies are commonly utilized as heating with boiled soy sauce, spreading soybean paste, or dipping into eggs, etc. Other drugs that have been employed are onion root, brine, eggplant, potato, loach, etc. The use of onion is based upon pain killing and antiphlogistic functions ; and that of brine upon antiphlogistic function. The use of soy sauce or soybean paste, fomentation, eggplant, potato and loach is said to be effective, but it has no pharmacblogic ground. 11) For the treatment of frostbite, such herb remedies are commonly used as dipping the affected part into frozen soybean sack, using boiled water of eggplant stem, garlic caulis, onion, hot pepper, caulis. Onion is based upon antiphlogistic and tranquility actions garlic upon disintection, metabolic exacerbation, tonic and aphrodisiac actions and the use of eggplant and hot pepper is based upon help blood circulation, dissolution and excretion of waste matters in vein. 12) For the treatment of burn, such herb remedies or drugs are commonly used as cleansing with Korean gin, spreading eggs, cleansing with cold water and soap water ; and as brine, cactus, moss, soybean paste, oil, etc. The cleansing with Korean gin, cold water, soap water, brine, vinegar is based upon cleaning and sterilizing functions ; and the use of cucumber is based upon nu. trition provision, and strengthening of resisting power by adjustment of metabolism. The use of potato, cactus, moss, oil and eggs is said to be effective, but their phamacological functions are not clarified. In view of the above results, we can realize that the drugs that have been employed in herb remedies are quite diverse. However, in regard to majority of herb remedies that have been employed by symptoms, the pharmacological functions of their drugs have not been clarified, and they are merely known as effective. Furthermore, they have not been recorded in the literature as yet ; and we confirm that there have been many herb remedies that were executed without the proper knowlege of their effects. It is now our view that the results of this survey may be utilized for consulting data in regard to the use of herb remedies.

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Effect of Gamijipaesan Extracts against Mastitis Induced by Staphylococcus aureus Infection in a Rat Model through Anti-inflammatory and Antibacterial Effects (가미지패산(加味芷貝散)의 포도상구균 감염 유방염에 대한 항균활성 및 항염 효과)

  • Kwon, Ji-Myung;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.1
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    • pp.1-24
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    • 2013
  • Objectives: The object of this study was to observe the protective effect of Gamijipaesan aqueous extracts(GJS), which has been traditionally used in Korean medicine in obstetrics & gynecological fields as anti-infectious and anti-inflammatory agents, against mastitis induced by Staphylococcus aureus infection in a rat model through antibacterial, antiinflammatory, immunomodulatory, and anti-oxidant effects. Methods: Antibacterial activities of GJS against S. aureus were detected using standard agar microdilution methods, with the effects on the bacterial invasion and intracellular killing of individual test materials in human mammary gland carcinoma cell(MCF-7) and murine macrophages(Raw 264.7) at MIC1/2, MIC and MIC2 concentration levels. In addition, the effects on the cell viability, nitric oxide(NO), tumor necrosis factor(TNF)-${\alpha}$ and interleukin (IL)-6 productions of LPS activated Raw 264.7 cells. The changes on the mammary tissue viable bacterial numbers, myeloperoxidae(MPO), inducible nitric oxide synthetase(iNOS), TNF-${\alpha}$ and IL-6 contents were observed in the S. aureus in vivo intramammary infectious rat model. The anti-bacterial and anti-inflammatory effects were compared with ciprofloxacin and piroxicam, respectively in the present study. Results: MIC of GJS and ciprofloxacin against S. aureus were detected as $0.860{\pm}0.428$ (0.391-1.563) mg/ml and $0.371{\pm}0.262$(0.098-0.782) ${\mu}g/ml$, respectively. In addition, GJS and ciprofloxacin were also showed marked dosage-dependent inhibition of the both bacterial invasion and intracellular killing assays using MCF-7 and Raw 264.7 cells at MIC1/2, MIC and $MIC{\times}2$ concentrations, respectively. $ED_{50}$ against LPS-induced cell viabilities and NO, TNF-${\alpha}$ and IL-6 releases of GJS were detected as 0.72, 0.04, 0.08 and 0.11 mg/ml, and as 19.04, 4.18, 5.37 and 4.27 ${\mu}g/ml$ in piroxicam, respectively. 250 and 500 mg/kg of GJS also inhibit the intramammary bacterial growth, MPO, iNOS, TNF-${\alpha}$ and IL-6 contents in S. aureus in vivo intramammary infected rats, respectively. GJS 500 mg/kg showed quite similar antibacterial and anti-infectious effects as compared with ciprofloxacin 40 mg/kg and also showed similar anti-inflammatory effects as piroxicam 10 mg/kg, in S. aureus in vivo intramammary infectious models. Conclusions: The results obtained in this study suggest that over 250 mg/kg of GJS showed favorable anti-infectious effects against S. aureus infection in a rat model through their antibacterial, anti-inflammatory, immunomodulatory and anti-oxidant effects and therefore expected that GJS can be used as alternative therapies, having both anti-inflammatory and anti-infectious activities. However, more detail mechanism studies should be conducted in future with the efficacy tests of individual herbal composition of GJS and the screening of the biological active compounds in individual herbs. In the present study, GJS 500 mg/kg showed quite similar anti-infectious effects were detected as compared with ciprofloxacin 40 mg/kg treated rats, and also GJS shows quite similar anti-inflammatory effects as compared with piroxicam 10 mg/kg in S. aureus in vivo intramammary infectious rats, but ciprofloxacin did not showed any anti-inflammatory effects, and piroxicam did not showed anti-infectious effects in this study.

Effects of Cyclobuxine E on Two Distinct Types of Potassium-Activated Calcium Channels in an Intestinal Smooth Muscle (흰쥐 장관에 있어 칼륨에 의해 활성화되는 칼슘 채널에 대한 Cyclobuxine E의 영향)

  • Lee, Jong-Hwoa;Kwon, Jun-Tack;Cho, Byung-Heon;Choi, Kyu-Hong;Kim, Yu-Jae;Kim, Jong-Bae;Kim, Chun-Sook;Cha, Young-Deog;Kim, Young-Suk
    • The Korean Journal of Pharmacology
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    • v.25 no.1
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    • pp.53-58
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    • 1989
  • For several years, we investigated the pharmacological action of several substances isolated from Buxus microphylla var koreana Nakai, which had been used as folk remedies of malaria and venereal disease. Cyclobuxine $D(C_{25}H_{42}ON_2)$, a steroidal alkaloid, exerted an antiinflammatory action, hypotensive and bradycardic effects in rats. In the present study, we isolated alkaloid from the acetone-insoluble fraction of the strong bases of this plants. This alkaloid $(C_{25}H_{38}ON_2)$ was identified as a steroidal alkaloid contained a cyclopropane ring by physical and chemical methods. It is a derivative of cyclobuxine D and named cyclobuxine E. We examined the effect of cyclobuxine E on the contractile response induced by acetylcholine and two distinct types of potassium-activated calcium channels in an intestinal smooth muscle of the rat. Cyclobuxine E inhibited significantly the Ach-induced contraction. The isolated longitudinal muscle from the rat duodenum was immersed calcium-depleted potassium depolarizing solution. Ten minutes after, 1.8 mM $CaCl_2$ was added to muscle bath and elicited a biphasic increase in muscle tension. Cyclobuxine E produced an appreciable inhibition of both components of the mechanical response. In addition, Cyclobuxine E introduced at a point when the tonic response had reached its maximum level, caused the muscle to exhibit a rapid loss of tension. Based on these experimental results, we proposed the possibility that the inhibitory action of cyclobuxine E on the isolated rat duodenum may be due to inhibiting the transmembrane fluxes of calcium ion in potassium-activated calcium channels.

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Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats (신생 백서의 저 산소 허혈 뇌손상에서 항세포사멸사를 통한 resveratrol의 신경보호 효과)

  • Shin, Jin Young;Seo, Min Ae;Choi, Eun Jin;Kim, Jin Kyung;Seo, Eok Su;Lee, Jun Hwa;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1102-1111
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    • 2008
  • Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.

TNF-α stimulated IL-8 and IL-10 expression in monocytes from patients with chronic granulomatous disease (만성육아종질환 환자 단핵구에서 TNF-α 자극에 의한 IL-8과 IL-10의 발현 양상)

  • Shin, Kyung-Sue
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1096-1101
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    • 2008
  • Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

A Comparative Study of Therapeutic Effect of Aspirin Alone and Intravenous Gammaglobulin Plus Aspirin in Kawasaki Disease (Kawasaki병에 있어 아스피린 단독 투여시와 정맥내 감마 글로불린 병용시에 치료효과에 대한 연구)

  • Hwang, Young-Hee;Jun, Jin-Gon;Kim, Chun-Dong
    • Journal of Yeungnam Medical Science
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    • v.9 no.1
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    • pp.44-53
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    • 1992
  • We compared the efficacy of each modality of treatment group in reducing the frequency of coronary artery abnormalities and changes of clinical courses in children with kawasaki disease in the children of 81 cases who were admitted in pediatric department of Yeungnam University Hospital from September 1985 to August 1990, Group A(37 cases)-aspirin alone, Group B(44 cases)-intravenous gammaglobulin(400mmg/kg/day) for 5 consecutive days, plus aspirin. We studied the frequency of echocardiographic abnormalities, the duration of fever, and changes in the total white blood cell counts, platelet counts, ESR and CRP value at 1, 2 and 3 weeks of the illness and compared the results between the two groups. The results were as follows, 1) There as no significant intergroup difference in age and sex ratio. 2) The duration of the febrile period after the initiation of the therapy was significant shorter in group B($2.5{\pm}1.2$ days) than in group A:($5.2{\pm}3.5$ days)(p<0.01). 3) No significant difference was noticed in the WBC and platelet counts in two groups as measured at admission day, 1 and 2 weeks of the illness. However, at 3weeks of illness significant difference was noted. 4) The CRP values measured at 1, 2 and 3 weeks after treatment were significantly lower in group B($2.42{\pm}1.8$, $2.00{\pm}1.2$, $1.16{\pm}1.0$) than in group A($7.22{\pm}5.3$, $5.25{\pm}3.9$, $1.85{\pm}1.2$) respectively(p<0.01). 5) In 2D- Echocardiogram, coronary artery dilataton was more frequent in Group A than in Group B at 6month of illness(p<0.01). In conclusion, intravenous gammaglobulin therapy was effective in the shortening of the duration of fever and in the antiinflammatory action and somewhat effective in prevention of coronary artery aneurysm.

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The Effects of Phenethyl Isothiocyanate on Nuclear Factor-κB Activation and Cyclooxygenase-2 and Inducible Nitric Oxide Synthase Expression Induced by Toll-like Receptor Agonists (Phenethyl Isothiocyanate가 Toll-like Receptor Agonists에 의해 유도된 Nuclear Factor-κB 활성과 Cyclooxygenase-2, Inducible Nitric Oxide Synthase 발현에 미치는 효과)

  • Kim, Soo-Jung;Park, Hye-Jeong;Shin, Hwa-Jeong;Kim, Ji-Soo;Ahn, Hee-Jin;Min, In-Soon;Youn, Hyung-Sun
    • Journal of Applied Biological Chemistry
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    • v.54 no.4
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    • pp.279-283
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    • 2011
  • Toll-like receptors (TLRs) play an important role in induction of innate immune responses. The activation of TLRs triggers inflammatory responses that are essential for host defense against invading pathogens. Phenethyl isothiocyanate (PEITC) extracted from cruciferous vegetables has an effect on anti-inflammatory therapy. Dysregulated activation of nuclear factor-${\kappa}$B (NF-${\kappa}$B), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) has been shown to play important roles in the development of certain inflammatory disease. To evaluate the therapeutic potential of PEITC, NF-${\kappa}$B activation and COX-2 and iNOS expression induced by lipopolysaccharide (LPS, TLR4 agonist), polyinosinic-polycytidylic acid (Poly[I:C], TLR3 agonist), 2 kDa macrophageactivating lipopeptide (MALP-2, TLR2 and TLR6 agonist) or oligodeoxynucleotide 1668 (ODN1668, TLR9 agonist) were examined. PEITC inhibits the activation of NF-${\kappa}$B induced by LPS or Poly[I:C] but not by MALP-2 or ODN1668. PEITC also suppressed the iNOS expression induced by LPS or Poly[I:C]. However, PEITC did not suppress COX-2 expression induced by LPS, Poly[I:C], MALP-2, or ODN1668. These results suggest that PEITC has the specific mechanism for antiinflammatory responses.

Combination Treatment with Arsenic Trioxide and Sulindac Induces Apoptosis of NCI-H157 Human Lung Carcinoma Cells via ROS Generation with Mitochondrial Dysfunction (NCI-H157 폐암 세포주에서 활성산소종의 생성과 미토콘드리아 기능변화를 한 Arsenic Trioxide와 Sulindac 병합요법의 세포고사효과)

  • Kim, Hak-Ryul;Yang, Sei-Hoon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.1
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    • pp.30-38
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    • 2005
  • Background : Arsenic trioxide ($As_2O_3$) has been used to treat acute promyelocytic leukemia, and it induces apoptosis in a variety of solid tumor cell lines including non-small cell lung cancer cells. However, nonsteroidal antiinflammatory drugs (NSAID) can enhance tumor response to chemotherapeutic drugs or radiation. It was previously demonstrated that a combination treatment with $As_2O_3$ and sulindac induces the apoptosis of NCI-H157 human lung carcinoma cells by activating the caspase cascade. This study aimed to determine if a combination treatment augmented its apoptotic potential through other pathways except for the activation of the caspase cascade. Material and Methods : The NCI-H157 cells were treated with $As_2O_3$, sulindac and antioxidants such as glutathione (GSH) and N-acetylcysteine (NAC). The cell viability was measured by a MTT assay, and the level of intracellular hydrogen peroxide ($H_2O_2$) generation was monitored fluorimetrically using a scopoletin-horse radish peroxidase (HRP) assay. Western blotting and mitochondrial membrane potential transition analysis were performed in order to define the mechanical basis of apoptosis. Results : The viability of the cells was decreased by a combination treatment of $As_2O_3$ and sulindac, and the cells were protected using antioxidants in a dose-dependent manner. The increased $H_2O_2$ generation by the combination treatment was inhibited by antioxidants. The combination treatment induced changes in the mitochondrial transmembrane potential as well as the expression of the Bcl-2 family proteins, and increased cytochrome c release into the cytosol. However, the antioxidants inhibited the effects of the combination treatment. Conclusion : Combination treatment with $As_2O_3$ and sulindac induces apoptosis in NCI-H157 human lung carcinoma cells via ROS generation with a mitochondrial dysfunction.