• Title/Summary/Keyword: BV2 microglial cells

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Inhibitory Effect of Chan-Su on the Secretion of PGE2 and NO in LPS-stimulated BV2 Microglial Cells

  • Kim, Min-Hee;Lyu, Ji-Hyo;Lyu, Sun-Ae;Hong, Sang-Hoon;Kim, Won-Il;Yoon, Hwa-Jung;Ko, Woo-Shin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.5
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    • pp.1315-1321
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    • 2008
  • Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulated BV2 microglial cells, Chan-Su inhibited the secretion of NO in BV2 microglial cells, without affecting cell viability, The protein level of inducible nitric oxide synthase (iNOS) was decreased by Chan-Su, And Chan-Su also inhibited production of prostaglandin E2 (PGE2) and expression of cyclooxygenase (COX)-2. Proinflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-12, were inhibited by Chan-Su in a dose-dependent manner. And Chan-Su inhibited the degradation of ${IkB-\alpha}$, which was considered to be inhibitor of nuclear factor $(NF)-{\kappa}B$, one of a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. These results suggest that Chan-Su could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $I{\kappa}B-{\alpha}$ degradation.

Inhibitory effects of Sam-Myo-San on the LPS-induced production of nitric oxide and $TNF-{\alpha}$ in RAW 264.7 cells and BV-2 Microglia cells (삼묘환(三妙丸)의 LPS에 의해 활성화된 RAW 264.7 cells과 BV-2 Microglia cells로부터 생성되는 nitric oxide 및 $TNF-{\alpha}$의 생성억제효과)

  • Lee, Jae-Hyun;Jung, Hyo-Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.59-67
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    • 2006
  • Objectives : Sam-Myo-Whan(SMW) has been known traditional prescription with anti- anthritis activities. We investigated inhibitory effects of SMW on lipopolysaccharide (LPS)-induced nitric oxide(NO), $TNF-{\alpha}$ and inducible nitric oxide synthase(iNOS) production from RAW264.7 cells and BV-2 Microglia cells. Methods : SMW, which had been extracted with 70% MeOH, concentrated and freeze-dried was used for this experiment. After BV2 mouse brain macrophages and RAW264.7 mouse peritoneal macrophages were pretreated with increasing concentrations of SMW extract for 30min, and then activated with LPS. To investigate cytotoxicity of SMW extract, cell viability was measured by MTT assay. NO production was measured in each culture supernatant by Griess reaction. mRNA expression of iNOS in two type cells was investigated by RT-PCR. $TNF-{\alpha}$ production was measured in each culture supernatant by ELISA. Results : SMW extract significantly inhibited LPS-induced NO and $TNF-{\alpha}$ production in BV2 cells and RAW264.7 cells dose-dependently. SMW extract also greatly suppressed mRNA expression of iNOS in both type cells activated with LPS. Conclusion : These data suggests that SMW extract may have an anti-inflammatory effect through the inhibition of iNOS expression.

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Inhibitory Effects of Asparagus cochinchinensis in LPS-Stimulated BV-2 Microglial Cells through Regulation of Neuroinflammatory Mediators, the MAP Kinase Pathway, and the Cell Cycle (Lipopolysaccharide로 자극된 BV-2 미세교세포에서 신경염증 매개체, MAP kinase경로, 세포주기의 조절에 의한 천문동(Asparagus cochinchinensis)의 저해효과)

  • Lee, Hyun Ah;Kim, Ji Eun;Choi, Jun Young;Sung, Ji Eun;Youn, Woo Bin;Son, Hong Joo;Lee, Hee Seob;Kang, Hyun-Gu;Hwang, Dae Youn
    • Journal of Life Science
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    • v.30 no.4
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    • pp.331-342
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    • 2020
  • The suppression of neuroinflammatory responses in microglial cells can be considered a key target for improving the progression of neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). Asparagus cochinchinensis has traditionally been used as a medicine to treat fever, cough, kidney disease, breast cancer, inflammatory diseases, and brain diseases. In this study, we investigated the neuroprotective mechanism of an aqueous extract from A. cochinchinensis root (AEAC), particularly its anti-inflammatory effects on lipopolysaccharide (LPS)-activated BV-2 microglial cells. BV-2 cells were treated with four different concentrations of AEAC. No significant toxicity was detected in BV-2 cells treated with AEAC. Nitric oxide (NO), cyclooxygenase-2 (COX-2) mRNA, and inducible nitric oxide synthase (iNOS) mRNA levels were 21% lower in the AEAC+LPS group than in the Vehicle+LPS group. Lower proinflammatory (TNF-α and IL-1β) and anti-inflammatory cytokine (IL-6 and IL-10) levels were also detected in the AEAC+LPS group than in the Vehicle+LPS group, albeit at varying rates. Moreover, the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group compared to the Vehicle+LPS group, enhancement of the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group, while cell cycle arrest at the G2/M phase caused by LPS treatment was less severe in the AEAC+LPS group. The increase in reactive oxygen species (ROS) generation induced by LPS treatment was also lower in the AEAC-pretreated group than in the Vehicle+LPS group. This is the first study to show that AEAC exerts anti-neuroinflammatory activity against LPS stimulation by regulating the MAPK signaling pathway, the cell cycle, and ROS production.

Anti-inflammatory activity of jakyakgamcho-tang on Lipopolysaccharide-Stimulated BV-2 Microglia Cells (LPS로 유도된 미세아교세포에서 작약감초탕의 항염증 효과)

  • Mun, Yeun-Ja
    • The Korea Journal of Herbology
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    • v.37 no.5
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    • pp.83-88
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    • 2022
  • Objectives : Jakyakgamcho-tang (JGT) has been traditionally used to treat muscular convulsion and pain in South Korea. According to recent studies, JGT has been reported to have anti-depression, anti-inflammation, anti-oxidative, anti-diabetics, anti-spasm and analgesic effects, but studies on its anti-neuroinflammatory and neuroprotective effect have not been deeply conducted. Thus, we investigated the anti-neuroinflammatory activity of JGT on lipopolysaccharide (LPS)-stimulated mouse microglia cells. Methods : To investigate the anti-neuroinflammatory effects of JGT on BV2 microglial cells, we examined the production of nitric oxide (NO) using griess assay, and mRNA expressions of pro-inflammatory cytokines such as interleukin (IL)-1𝛽, IL-6, and tumor necrosis factor (TNF)-𝛼 using real time RT-PCR. Furthermore, to determine the regulating mechanisms of JGT, we investigated the heme oxygenase (HO)-1 by real time RT-PCR. Results : Pre-treatment of JGT effectively decreased NO production in LPS-stimulated BV2 cells at concentrations without cytotoxicity. Additionally, JGT significantly suppressed the production of IL-1𝛽, IL-6, and TNF-𝛼 in LPS-stimulated BV2 cells. Furthermore, JGT activated the HO-1 expression, which is one of the immunomodulatory signaling molecules. And the abolishment of HO-1 by tin protoporphyrin IX (SnPP, the HO-1 inhibitor) reversed the anti- inflammatory activity of JGT in LPS-stimulated BV2 cells. Conclusions : Our results suggest that the JGT has anti-neuroinflammatory effect through the activation of HO-1 in LPS-stimulated BV2 cells. Thereby, JGT could expected to be used for the prevention and treatment of neurodegenerative disease related to neuroinflammation.

Effects of Armeniacae Semen and Amygdalin on Prostaglandin E2 Synthesis and Nitric Oxide Production (행인(杏仁)과 Amygdalin이 Prostaglandin E2 합성과 NO생성에 미치는 영향)

  • Jung, Hyung-Jin;Kim, Youn-Sub;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.32 no.3
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    • pp.13-22
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    • 2019
  • Objectives : Armeniacae semen is the seed of Prunus armenica L. var. ansu MAXIM, and this is classified into Rosaceae. Armeniacae semen has been used for centuries in traditional oriental medicine for the treatment of pain and inflammatory diseases. Amygdalin is the major compound of Armeniacae semen, and it is now being used for the treatment of pain and cancer. Methods : In the present study, we compared the effects of an aqueous extract of Armeniacae semen and a solution of amygdalin extracted from Armeniacae semen on lipopolysaccharide(LPS)-stimulated prostaglandin E2 synthesis and nitric oxide production in mouse BV-2 microglial cells. For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, reverse transcription-polymerase chain reaction(RT-PCR), prostaglandin E2 immunoassay and nitric oxide detection were performed on mouse BV-2 microglial cells. Results : In the present study, an aqueous extract of Armeniacae semen and an amygdalin solution extracted from Armeniacae semen suppressed prostaglandin E2 synthesis and nitric oxide production by inhibiting the LPS-induced enhancement of cyclooxygenase-2(COX-2) mRNA and the inducible nitric oxide synthase mRNA in mouse BV-2 cells. For the cyclooxygenase-1(COX-1) expression, an aqueous extract of Armeniacae semen showed a more potent suppression effect compared to the amygdalin solution. However, the amygdalin solution more potently suppressed the LPS-induced COX-2 mRNA expression compared to the aqueous extract of Armeniacae semen. Conclusions : As a result, aqueous extract of Armeniacae semen and amygdalin exert anti-inflammatory and analgesic effects.

The Effects of Bee Venom Pharmacopuncture on Middle Cerebral Artery Occlusion Ischemic Cerebral Damage in Mice

  • Lee, Ji-In;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.36 no.4
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    • pp.220-229
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    • 2019
  • Background: The therapeutic potential of Bee Venom Pharmacopuncture (BVP) on acute ischemic cerebral infraction was determined in mice in vivo and in vitro. Methods: Analysis of acute ischemic cerebral infraction was performed using 7 week old male ICR mice (n = 20) and microglial BV-2 cells. Bee venom ($5{\mu}g/kg$) was injected into the caudal vein of middle cerebral artery occlusion (MCAo) mice (1 hour after reperfusion, 3 hours after MCAo probe insertion), and also used to treat LPS-stimulated microglial BV-2 cells (1, 2, $5{\mu}g/mL$). Markers of inflammation were monitored. Results: NO declined statistically significantly in BVP treated MCAo mice compared to the untreated MCAo group (p < 0.05). Compared to the MCAo group, the BVP-treated MCAo group showed a decreased production volume of malondialdehyde, but an increased glutathione/oxidized glutathione ratio. Compared to the untreated MCAo group, the BVP treated MCAo group showed a statistically significant decline in TNF and $IL-1{\beta}$ levels (p < 0.05). BVP inhibited the levels of p65, p50, $p-I{\kappa}B-{\alpha}$, and levels of p-ERK1/2, p-JNK2, p-P38 declined. Conclusion: BVP is effective at dampening the inflammatory response in vivo and in vitro and may supplement rt-PA treatment.

Effects of Lesser Yang Person-Hyungbangpaedok-san on Lipopolysaccharide-induced Expressions of cyclooxygenase-2 and Inducible Nitric Oxide Synthase in Mouse BV2 Microglial Cells (소양인형방패독산의 생쥐 소교세포에서 지질다당질에 의한 cyclooxygenage-2와 inducible nitric oxide synthase 발현에 미치는 영향)

  • Koh, Ji-Eun;Kim, Jullia;Kim, Hong;Kim, Chang-Ju;Lee, Choong-Yeol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.4
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    • pp.961-966
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    • 2007
  • Lesser yang person-Hyungbangpaedok-san (HBPDS) is a prescription originated in the

Effects of the Aqueous Extract of Rehmanniae Radix Preparata on Lipopolysaccharide-induced Expressions of Cyclooxygenase-2 and Inducible Nitric Oxide Synthase in Mouse BV2 Microglial Cells

  • Jung, Chang-Young;Sung, Yun-Hee;Kim, Sung-Eun;Kim, Chang-Ju;Han, Seung-Ho;Lee, Choong-Yeol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.4
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    • pp.1051-1056
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    • 2006
  • Rehmanniae radix preparata is the root of Rehmanniae glutinosa Liboschitz var. purpurea Makino which has been classified into Scrophulariaceae. Rehmanniae radix preparata has been used for the treatment of diabetes, for the relief of the pain, and for the anti-oxidative action. In this study, the effect of the aqueous extract of Rehmanniae radix preparata on lipopolysaccharide-induced inflammation was investigated by using 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, prostaglandin E2 immunoassay, and nitric oxide (NO) detection in mouse BV2 microglial cells. In the present results, the aqueous extract of Rehmanniae radix preparata suppressed prostaglandin E2 (PGE2) synthesis and nitric oxide production by inhibiting the lipopolysaccharide-stimulated expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) mRNA and protein in mouse BV2 cells. These results show that Rehmanniae radix preparata exerts anti-inflammatory effect probably by suppressing of COX-2 and iNOS expressions.

Solanum nigrum L. Extract Inhibits Inflammation in Lipopolysaccharide-stimulated Raw 264.7 and BV2 Cells

  • Lee, Jin Wook;Jung, Hyuk-Sang;Sohn, Youngjoo;Kang, Yoon Joong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.92-92
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    • 2018
  • Solanum nigrum L. (SNL), generally known as black nightshade, is traditionally used as medicine to reduce inflammation caused by several diseases like asthma, chronic bronchitis and liver cirrhosis. In this study, anti-inflammatory effects of SNL extract were examined and possible molecular mechanisms of the anti-inflammatory effects were investigated. The inhibitory effects of SNL extract on nitric oxide (NO), pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6) and Matrix metallopeptidase 9 (MMP-9) productions were dissected using lipopolysaccharide (LPS) stimulated murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. We further investigated whether SNL extract could suppress the phosphorylation of ERK1/2, JNK, and p38 and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 in LPS-stimulated Raw264.7 cells and BV2 cells. As a result, we showed that the SNL extract significantly decreased the production of pro-inflammatory cytokines, NO, and MMP-9. In addition, the SNL strongly inhibited the phosphorylation of ERK1/2, JNK, p38 and nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. We confirmed that the extracts of SNL effectively inhibits the anti-inflammatory and may be used as a therapeutic to various inflammatory diseases.

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Chloroform Fraction of Zingiberis Rhizoma Recens Modulates the Production of Inflammatory Mediators in LPS-stimulated BV2 Microglial Cells (생강 클로로포름 분획의 활성화된 뇌신경교세포(腦神經膠細胞)에서 염증반응 억제효과)

  • Seo, Un-Kyo;Jung, Hyo-Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.73-83
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    • 2008
  • Objectives : The root of Zingiber officinale ROSC. (Zingiberis Rhizoma Recens; Ginger) has been widely used as one of folk remedies and food materials in many traditional preparations. Ginger is known as an effective appetite enhancer and anti-inflammatory agent. This study was performed to investigate the effect of ginger chloroform fraction (GCF) in microglia which play a central role on brain inflammation in neurodegenerative diseases. Methods : Dried ginger was extracted with 80% methanol, and then fractionated with chloroform. BV2 mouse microglial cells were cultured with different concentrations of GCF and then stimulated with LPS (1 ${\mu}g/m{\ell}$) at indicated times. The cell toxicity of GCF was determined by MTT assay. The concentrations of NO, PGE2 and cytokines were measured by Griess assay and enzyme-linked immunosorbant assay. The mRNA and protein expressions of iNOS, COX-2 and cytokines were determined by RT-PCR and Western blotting. The phosphorylation of three MAPKs (p38 MAPK, ERK1/2 and JNK) and $NF-{\kappa}B$ activation were determined by Western blotting. Results : GCF significantly inhibited LPS-induced production of inflammatory mediators, NO, $PGE_2$ and proinflammatory cytokines ($TNF-{\alpha}$ and $IL-1{\beta}$) in a dose-dependent manner. GCF attenuated LPS-induced expression of mRNA and protein of inflammatory enzymes, iNOS, COX-2 and proinflammatory cytokines through suppressing the phosphorylation of ERK1/2 and p38 MAPK and the activation of p65 $NF-{\kappa}B$ in BV2 cells. Conclusions : This study suggests that GCF may have an anti-inflammatory property through suppressing the inflammatory mediator production released by activated microglia after the brain injury.

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