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Role of Protein Kinases on NE-$_{\kappa}B$ Activation and Cell Death in Bovine Cerebral Endothelial Cells

  • Ahn, Young-Soo;Kim, Chul-Hoon;Kim, Joo-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • 제3권1호
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    • pp.11-18
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    • 1999
  • Nuclear factor $_{\kappa}B\;(NF-_{\kappa}B)$ activation is modulated by various protein kinases. Activation of $NF-_{\kappa}B$ is known to be important in the regulation of cell viability. The present study investigated the effect of inhibitors of protein tyrosine kinase (PTK), protein kinase C (PKC) and protein kinase A (PKA) on $NF-_{\kappa}B$ activity and the viability of bovine cerebral endothelial cells (BCECs). In serum-deprivation-induced BCEC death, low doses of $TNF{\alpha}$ showed a protective effect. $TNF{\alpha}$ induced $NF-_{\kappa}B$ activation within 4 h in serum-deprivation. PTK inhibitors (herbimycin A and genistein) and PKC inhibitor (calphostin C) prevented $NF-_{\kappa}B$ activation stimulated by $TNF{\alpha}.$ Likewise, these inhibitors prevented the protective effect of $TNF{\alpha}.$ In contrast to $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activity, basal $NF-_{\kappa}B$ activity of BCECs in media containing serum was suppressed only by calphostin C, but not by herbimycin A. As well BCEC death was also induced only by calphostin C in serum-condition. H 89, a PKA inhibitor, did not affect the basal and $TNF{\alpha}-stimulated\;NF-_{\kappa}B$ activities and the protective effect of $TNF{\alpha}$ on cell death. These data suggest that modulation of $NF-_{\kappa}B$ activation could be a possible mechanism for regulating cell viability by protein kinases in BCECs.

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청피의 항염증효과 (Effects of Citri Reticulatae Viride Pericarpium on 4-Hydroxynonenal-Induced Inflammation in PC12 Cells)

  • 예영준;김연섭;강미숙
    • 한방비만학회지
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    • 제16권2호
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    • pp.79-84
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    • 2016
  • Objectives: The purpose of this study was to observe the effects of Citri Reticulatae Viride Pericarpium (CP) on 4-Hydroxynonenal (4-HNE)-induced inflammation in PC12 cells. Methods: 4-HNE was treated in PC12 cell to cause inflammatory response, and then treated with CP water extract at 25, 50, and $100{\mu}g/ml$. The phosphorylation of Jun N-terminal kinase (JNK) and the expression of $NF-{\kappa}B$ in PC12 cells were determined by Western blot, respectively. Results: The phosphorylation of JNK was significantly decreased in 4-HNE-stimulated PC12 cell by the treatment of CP extract at $25{\mu}g/ml$. The 4-HNE-induced expression of nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) p65 in nuclear of the cells was significantly decreased in PC12 cell by treatment with CP extract at 25, 50, and $100{\mu}g/ml$. Conclusions: These results suggest that CP water extract has an anti-inflammatory activity through suppressing the JNK and $NF-{\kappa}B$ activation.

가감양격산(加減凉膈散)이 RBL-2H3 비만세포와 OVA/alum에 감작된 생쥐에 미치는 항알레르기 효과 (Anti-allergic Effects of Gagam-YangGyeokSan on RBL-2H3 Mast Cells and OVA/alum Sensitized Mice)

  • 이윤실;한재경;김윤희
    • 대한한방소아과학회지
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    • 제26권4호
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    • pp.10-23
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    • 2012
  • Objectives: Gagamyanggyeoksan (G-YGS) has been used to suppress allergic reaction, however, the cellular target of G-YGS and its mode of action remain unclear. The present study was designed to investigate the effect of extracted G-YGS on the PMA and lonomycin (PI)-induced activation of RBL-2H3. Methods: For this investigation, We examined IL-4, IL-13 mRNA expression by Real-Time PCR, IL-4, IL-13 production by ELISA analysis and manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting, OVA-specific IgE, IL-4, IL-13 by mouse be sensitive to OVA. Results: Here we showed that treatment of RBL-2H3 mast cells with G-YGS, suppressed PI-induced production of Th2 cytokines including IL-4 and IL-13 in a dose dependent manner. The mRNA expression of IL-4 were completely abolished by G-YGS at the concentration of $100{\mu}g/ml$. Data from a stable cell lines consistently expressing IL-4. And the mRNA expression of IL-13 were abolished by G-YGS at the $200{\mu}g/ml$. But there is no difference between the $50{\mu}g/ml$, the $100{\mu}g/ml$ and the comparison. Results from the western blot analysis of transcription factors involving IL-4 and IL-13 expression indicated that it prominently decreased the expression of mast cell specific transcricption factors including GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65 but not c-Fos. And G-YGS suppressed IgE, IL-4, IL-13 in mouse be sensitive to OVA. Conclusions We suggested the anti-allergic activities of G-YGS might be mediated by down-regulation of Th2 cytokines such as IL-4 and IL-13 through the regulation of transcription factors as GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65.

Salsolinol, a tetrahydroisoquinoline-derived neurotoxin, induces oxidative modification of neurofilament-L: protection by histidyl dipeptides

  • Kang, Jung-Hoon
    • BMB Reports
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    • 제45권2호
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    • pp.114-119
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    • 2012
  • Salsolinol (1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) is a compound derived from dopamine metabolism and is capable of causing dopaminergic neurodegeneration. Oxidative modification of neurofilament proteins has been implicated in the pathogenesis of neurodegenerative disorders. In this study, oxidative modification of neurofilament-L (NF-L) by salsolinol and the inhibitory effects of histidyl dipeptides on NF-L modification were investigated. When NF-L was incubated with 0.5 mM salsolinol, the aggregation of protein was increased in a time-dependent manner. We also found that the generation of hydroxyl radicals (${\bullet}OH$) was linear with respect to the concentrations of salsolinol as a function of incubation time. NF-L exposure to salsolinol produced losses of glutamate, lysine and proline residues. These results suggest that the aggregation of NF-L by salsolinol may be due to oxidative damage resulting from free radicals. Carnosine, histidyl dipeptide, is involved in many cellular defense processes, including free radical detoxification. Carnosine, and anserine were shown to significantly prevent salsolinol-mediated NF-L aggregation. Both compounds also inhibited the generation of ${\bullet}OH$ induced by salsolinol. The results indicated that carnosine and related compounds may prevent salsolinol-mediated NF-L modification via free radical scavenging.

USP15 inhibits multiple myeloma cell apoptosis through activating a feedback loop with the transcription factor NF-κBp65

  • Zhou, Lili;Jiang, Hua;Du, Juan;Li, Lu;Li, Rong;Lu, Jing;Fu, Weijun;Hou, Jian
    • Experimental and Molecular Medicine
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    • 제50권11호
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    • pp.11.1-11.12
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    • 2018
  • USP15 has been shown to stabilize transcription factors, to be amplified in many cancers and to mediate cancer cell survival. However, the underlying mechanism by which USP15 regulates multiple myeloma (MM) cell proliferation and apoptosis has not been established. Here, our results showed that USP15 mRNA expression was upregulated in MM patients. USP15 silencing induced MM cell proliferation inhibition, apoptosis, and the expression of nuclear and cytoplasmic NF-${\kappa}Bp65$, while USP15 overexpression exhibited an inverse effect. Moreover, in vivo experiments indicated that USP15 silencing inhibited MM tumor growth and NF-${\kappa}Bp65$ expression. PDTC treatment significantly inhibited USP15 overexpression-induced cell proliferation, apoptosis inhibition, and NF-${\kappa}Bp65$ expression. USP15 overexpression promoted NF-${\kappa}Bp65$ expression through inhibition of its ubiquitination, whereas NF-${\kappa}Bp65$ promoted USP15 expression as a positive regulator. Taken together, the USP15-NF-${\kappa}Bp65$ loop is involved in MM tumorigenesis and may be a potential therapeutic target for MM.

6-Shogaol의 Toll-like receptor 2, 3, 4 agonists에 의해서 유도된 cyclooxygenase-2 발현 억제 (Suppression of the Expression of Cyclooxygenase-2 Induced by Toll-like Receptor 2, 3, and 4 Agonists by 6-Shogaol)

  • 김점지;안상일;이전수;윤새미;이미영;윤형선
    • 한국식품과학회지
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    • 제40권3호
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    • pp.332-336
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    • 2008
  • 선천성 면역은 병원성균의 침입에 대항하기 위한 숙주의 최초 방어체계라 할 수 있다. 이러한 선천성 면역반응은 병원균들이 가지고 있는 독특한 구조를 인식하는 TLRs에 의해서 조절되어 진다고 알려져 있다. 숙주에 침입한 여러 병원성균들이 TLRs를 자극하며 이렇게 자극된 신호들은 아래로 전달되어 전사요소 $NF-{\kappa}B$의 활성화를 유도하고 결국 COX-2와 같은 염증 유발인자를 유도하여 암이나 질병을 유발하게 된다. 우리는 이번 연구를 통하여 생강 추출물중의 하나인 6-shogaol이 어떻게 $NF-{\kappa}B$ 활성화나 COX-2 발현을 조절하여 항염증 효과를 가지고 있는지를 알아보았다. 6-shogaol은 TLR2, TLR3, TLR4 agonists에 의해서 유도된 $NF-{\kappa}B$ 활성화와 COX-2 발현을 억제하였다. 이러한 결과는6-shogaol이 여러 병원균들로부터 유도되는 염증반응이나 만성적인 질병들을 조절할 수 있다는 중요한 결과를 보여주는 것이라 할 수 있다.

Toll-like receptor 4/nuclear factor-kappa B pathway is involved in radicular pain by encouraging spinal microglia activation and inflammatory response in a rat model of lumbar disc herniation

  • Zhu, Lirong;Huang, Yangliang;Hu, Yuming;Tang, Qian;Zhong, Yi
    • The Korean Journal of Pain
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    • 제34권1호
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    • pp.47-57
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    • 2021
  • Background: Lumbar disc herniation (LDH) is a common cause of radicular pain, but the mechanism is not clear. In this study, we investigated the engagement of toll-like receptor 4 (TLR4) and the nuclear factor-kappa B (NF-κB) in radicular pain and its possible mechanisms. Methods: An LDH model was induced by autologous nucleus pulposus (NP) implantation, which was obtained from coccygeal vertebra, then relocated in the lumbar 4/5 spinal nerve roots of rats. Mechanical and thermal pain behaviors were assessed by using von Frey filaments and hotplate test respectively. The protein level of TLR4 and phosphorylated-p65 (p-p65) was evaluated by western blotting analysis and immunofluorescence staining. Spinal microglia activation was evaluated by immunofluorescence staining of specific relevant markers. The expression of proand anti-inflammatory cytokines in the spinal dorsal horn was measured by enzyme linked immunosorbent assay. Results: Spinal expression of TLR4 and p-NF-κB (p-p65) was significantly increased after NP implantation, lasting up to 14 days. TLR4 was mainly expressed in spinal microglia, but not astrocytes or neurons. TLR4 antagonist TAK242 decreased spinal expression of p-p65. TAK242 or NF-κB inhibitor pyrrolidinedithiocarbamic acid alleviated mechanical and thermal pain behaviors, inhibited spinal microglia activation, moderated spinal inflammatory response manifested by decreasing interleukin (IL)-1β, IL-6, tumor necrosis factor-α expression and increasing IL-10 expression in the spinal dorsal horn. Conclusions: The study revealed that TLR4/NF-κB pathway participated in radicular pain by encouraging spinal microglia activation and inflammatory response.

한국산 꾸구리속, Gobiobotia (Pisces: Cyprinidae) 3종의 핵형 (Karyotypes of Three Species of Gobiobotia (Pisces: Cyprinidae) in Korea)

  • 송호복;박갑만
    • 한국어류학회지
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    • 제17권3호
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    • pp.159-166
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    • 2005
  • 한국산 꾸구리속 (Gobiobotia) 어류인 꾸구리 (G. macrocephala), 돌상어 (G. brevibarba) 및 흰수마자 (G. nakdongensis)의 핵형을 조사하였다. 연구 결과, 각각 25쌍의 배수체 염색체가 관찰되었 다. 핵형은 꾸구리가 2n = 50 (9M+7SM+9ST), NF = 100, 염색체의 길이는 $3.3{\sim}7.5{\mu}m$ 크기였으며, 돌상어는 2n = 50 (10M+7SM+4ST+4T), NF = 92, 염색체의 길이는 $2.7{\sim}6.3{\mu}m$의 크기였고, 흰수마자는 2n = 50 (5M+9SM+9ST+2T), NF = 96, 염색체의 길이는 $3.5{\sim}7.3{\mu}m$였다. 본 연구결과 꾸구리와 흰수마자의 핵형은 처음으로 보고되었다.

Licochalcone B Exhibits Anti-inflammatory Effects via Modulation of NF-κB and AP-1

  • Kim, Jin-Kyung;Jun, Jong-Gab
    • 대한의생명과학회지
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    • 제21권4호
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    • pp.218-226
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    • 2015
  • The present study investigated the mechanisms of licochalcone B (LicB)-mediated inhibition of the inflammatory response in murine macrophages. RAW264.7 murine macrophages were cultured in the absence or presence of lipopolysacharide (LPS) with LicB. LicB suppressed the generation of nitric oxide and the pro-inflammatory cytokines interleukin (IL)-$1{\beta}$, IL-6 and tumor necrosis factor-${\alpha}$. LicB also inhibited the expression of mRNA for inducible nitric oxide synthase and pro-inflammatory cytokines induced by LPS. Moreover, LicB inhibited nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and activator protein-1 translocation into the nucleus in a dose-dependent manner. Thus, LicB mainly exerts its anti-inflammatory effects by inhibiting the LPS-induced NF-${\kappa}B$ and activator protein-1 signaling pathways in macrophages, which subsequently diminishes the expression and release of various inflammatory mediators. LicB shows promise as a therapeutic agent in inflammatory diseases.

Application of nanofiltration membrane for the River Nile water treatment in Egypt: Case study

  • Jamil, Tarek S.;Shaban, Ahmad M.;Mansor, Eman S.;Karim, Ahmed A.;El-Aty, Azza M. Abd
    • Membrane and Water Treatment
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    • 제9권4호
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    • pp.233-243
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    • 2018
  • In this manuscript, $35m^3/d$ NF unit was designed and applied for surface water treatment of the River Nile water. Intake of Embaba drinking water treatment plant was selected to install that unit at since; it has the lowest water quality index value through the examined 6 sites in greater Cairo area. The optimized operating conditions were feed and permeate flow, 40 and $7m^3/d$, feed pressure 2.68 bar and flux rate $37.7l/m^2h$. The permeate water was drinkable according to Egyptian Ministerial decree 458/2007 for the tested parameters (physic-chemical, heavy metals, organic, algal, bacteriological and parasitological). Single and double sand filters were used as pretreatment for NF membranes but continuous clogging for sand filters moved us to use UF membrane as pretreatment for NF membrane.