scholarly journals Cannabinoid CB2 Receptor Mediates Nicotine-Induced Anti-Inflammation in N9 Microglial Cells Exposed toβAmyloid via Protein Kinase C

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Ji Jia ◽  
Jie Peng ◽  
Zhaoju Li ◽  
Youping Wu ◽  
Qunlin Wu ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Microglial Activation ◽  
Microglial Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cb2 Receptor ◽  
Arginase 1 ◽  
Kinase C ◽  
Activated State ◽  
Anti Inflammation

Background. Reducingβamyloid- (Aβ-) induced microglial activation is considered to be effective in treating Alzheimer’s disease (AD). Nicotine attenuates Aβ-induced microglial activation; the mechanism, however, is still elusive. Microglia could be activated into classic activated state (M1 state) or alternative activated state (M2 state); the former is cytotoxic and the latter is neurotrophic. In this investigation, we hypothesized that nicotine attenuates Aβ-induced microglial activation by shifting microglial M1 to M2 state, and cannabinoid CB2 receptor and protein kinase C mediate the process.Methods. We used Aβ1–42 to activate N9 microglial cells and observed nicotine-induced effects on microglial M1 and M2 biomarkers by using western blot, immunocytochemistry, and enzyme-linked immunosorbent assay (ELISA).Results. We found that nicotine reduced the levels of M1 state markers, including inducible nitric oxide synthase (iNOS) expression and tumor necrosis factorα(TNF-α) and interleukin- (IL-) 6 releases; meanwhile, it increased the levels of M2 state markers, including arginase-1 (Arg-1) expression and brain-derived neurotrophic factor (BDNF) release, in the Aβ-stimulated microglia. Coadministration of cannabinoid CB2 receptor antagonist or protein kinase C (PKC) inhibitor partially abolished the nicotine-induced effects.Conclusion. These findings indicated that cannabinoid CB2 receptor mediates nicotine-induced anti-inflammation in microglia exposed to Aβvia PKC.

scholarly journals SOCS1 Mediates Berberine-Induced Amelioration of Microglial Activated States in N9 Microglia Exposed to β Amyloid

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Qi Guo ◽  
Chen Wang ◽  
Xiaorong Xue ◽  
Bin Hu ◽  
He Bao
Keyword(s):  
Neurotrophic Factor ◽  
Microglial Activation ◽  
Microglial Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cytokine Signaling ◽  
Arginase 1 ◽  
Inos Protein Expression ◽  
Activated State ◽  
Β Amyloid ◽  
Factor Α

Attenuating β amyloid- (Aβ-) induced microglial activation is considered to be effective in treating Alzheimer’s disease (AD). Berberine (BBR) can reduce microglial activation in Aβ-treated microglial cells; the mechanism, however, is still illusive. Silencing of cytokine signaling factor 1 (SOCS1) is the primary regulator of many cytokines involved in immune reactions, whose upregulation can reverse the activation of microglial cells. Microglia could be activated into two different statuses, classic activated state (M1 state) and alternative activated state (M2 state), and M1 state is harmful, but M2 is beneficial. In the present study, N9 microglial cells were exposed to Aβ to imitate microglial activation in AD. And Western blot and immunocytochemistry were taken to observe inducible nitric oxide synthase (iNOS), Arginase-1 (Arg-1), and SOCS1 expressions, and the enzyme-linked immunosorbent assay (ELISA) was used to measure inflammatory and neurotrophic factor release. Compared with the normal cultured control cells, Aβ exposure markedly increased the level of microglial M1 state markers ( P < 0.05 ), including iNOS protein expression, tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 releases, and BBR administration upregulated SOSC1 expression and the level of microglial M2 state markers ( P < 0.05 ), such as Arg-1 expression, brain-derived neurotrophic factor (BDNF), and glial cell-derived neurotrophic factor (GDNF) releases, downregulating the SOCS1 expression by using siRNA, however, significantly reversed the BBR-induced effects on microglial M1 and M2 state markers and SOCS1 expression ( P < 0.05 ). These findings indicated that BBR can inhibit Aβ-induced microglial activation via modulating the microglial M1/M2 activated state, and SOCS1 mediates the process.

scholarly journals Delayed Treatment with Isoflurane Attenuates Lipopolysaccharide and Interferon γ–induced Activation and Injury of Mouse Microglial Cells

2009 ◽  
Vol 111 (3) ◽  
pp. 566-573 ◽  
Author(s):  
Jie-Ae Kim ◽  
Liaoliao Li ◽  
Zhiyi Zuo
Keyword(s):  
Protein Kinase C ◽  
Lactate Dehydrogenase ◽  
Protein Kinase ◽  
Cell Viability ◽  
Inos Expression ◽  
Microglial Cells ◽  
Lactate Dehydrogenase Release ◽  
Kinase C ◽  
Nitrite Production ◽  
Isoflurane Treatment

Background Isoflurane pretreatment can induce protection against lipopolysaccharide and interferon gamma (IFNgamma)-induced injury and activation of mouse microglial cells. This study's goal was to determine whether delayed isoflurane treatment is protective. Methods Mouse microglial cells were exposed to various concentrations of isoflurane for 1 h immediately after the initiation of lipopolysaccharide (10 or 1000 ng/ml) and IFNgamma (10 U/ml) stimulation or to 2% isoflurane for 1 h at various times after initiation of the stimulation. Nitrite production, lactate dehydrogenase release, and cell viability measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were assessed after stimulation with lipopolysaccharide and IFNgamma for 24 h. Inducible nitric oxide synthase (iNOS) protein expression was quantified by Western blotting. The iNOS expression in mouse brain was also studied. Results Isoflurane applied 0 and 2 h after the initiation of lipopolysaccharide and IFNgamma stimulation improved cell viability. Isoflurane at 2%, but not at 1 or 3%, reduced the lipopolysaccharide and IFNgamma-induced nitrite production and decreased cell viability. Aminoguanidine, an iNOS inhibitor, also attenuated this decreased cell viability. Chelerythrine and bisindolylmalemide IX, protein kinase C inhibitors, abolished isoflurane effects on cell viability and iNOS expression after lipopolysaccharide and IFNgamma application. Isoflurane also decreased lipopolysaccharide-induced iNOS expression in mouse brain. Late isoflurane application to microglial cells reduced lipopolysaccharide and IFNgamma-induced lactate dehydrogenase release that was not inhibited by aminoguanidine. Conclusions These results suggest that delayed isoflurane treatment can reduce lipopolysaccharide and IFNgamma-induced activation and injury of microglial cells. These effects may be mediated by protein kinase C.

scholarly journals Properties and distribution of the protein inhibitor (Mr 17,000) of protein kinase C

1987 ◽  
Vol 242 (3) ◽  
pp. 695-705 ◽  
Author(s):  
J R McDonald ◽  
U Gröschel-Stewart ◽  
M P Walsh
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Hydrophobic Interaction ◽  
Binding Proteins ◽  
Proteinase Inhibitors ◽  
Hydrophobic Interaction Chromatography ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Proteolytic Fragment ◽  
Kinase C

Ca2+-dependent hydrophobic-interaction chromatography is a powerful tool for the identification and isolation of a variety of Ca2+-binding proteins which expose a hydrophobic site(s) in the presence of Ca2+ [Gopalakrishna & Anderson (1982) Biochem. Biophys. Res. Commun. 104, 830-836; Walsh, Valentine, Ngai, Carruthers & Hollenberg (1984) Biochem. J. 224, 117-127; McDonald & Walsh (1985) Biochem. J. 232, 559-567]. Using this approach, we isolated two potent and specific protein inhibitors of protein kinase C, of 17 kDa [McDonald & Walsh (1985) Biochem. J. 232, 559-567] and 12 kDa [McDonald & Walsh (1986) Biochem. Soc. Trans. 14, 585-586]. Although these inhibitors were purified by Ca2+-dependent hydrophobic-interaction chromatography and exhibit properties similar to those of calmodulin and related Ca2+-binding proteins, we were unable to demonstrate high-affinity Ca2+ binding to these inhibitors, using equilibrium dialysis. Protein kinase C exhibited half-maximal activity at 0.6 microM-Ca2+ in the presence of phospholipid and diacylglycerol, and complete inhibition by both inhibitors was observed over the range of Ca2+ concentrations examined (10 nM-10 microM). These observations suggest that the inhibitory action of these proteins does not require Ca2+. The inclusion of proteinase inhibitors during isolation of the kinase C inhibitors, as well as two-dimensional peptide mapping and amino acid analysis of the isolated proteins, suggested that the 12 kDa inhibitor is a proteolytic fragment of the 17 kDa protein which is generated during purification. Antibodies raised in rabbits against the bovine brain 17 kDa inhibitor were shown to be specific by Western immunoblotting and the competitive enzyme-linked immunosorbent assay method and were used to study the tissue and species distribution of this protein. The inhibitor was found to be present in several bovine, murine, avian and human tissues, consistent with a role in the regulation of a variety of physiological functions involving the widely distributed protein kinase C.

scholarly journals Gypenoside AttenuatesβAmyloid-Induced Inflammation in N9 Microglial Cells via SOCS1 Signaling

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Hui Cai ◽  
Qianlei Liang ◽  
Guanqun Ge
Keyword(s):  
Neurotrophic Factor ◽  
Microglial Activation ◽  
Interleukin 1 ◽  
Chinese Herb ◽  
Microglial Cells ◽  
Amyloid A ◽  
Arginase 1 ◽  
Traditional Chinese Herb ◽  
Activated State ◽  
Necrosis Factor

Reducingβamyloid- (Aβ-) induced microglial activation is believed to be effective in treating Alzheimer’s disease (AD). Microglia can be activated into classic activated state (M1 state) or alternative activated state (M2 state), and the former is harmful; in contrast, the latter is beneficial. Gypenoside (GP) is the major bioactive constituent ofGynostemma pentaphyllum, a traditional Chinese herb medicine. In this study, we hypothesized that GP attenuates Aβ-induced microglial activation by ameliorating microglial M1/M2 states, and the process may be mediated by suppressor of cell signaling protein 1 (SOCS1). In this study, we found that Aβexposure increased the levels of microglial M1 markers, including iNOS expression, tumor necrosis factorα(TNF-α), interleukin 1β(IL-1β), and IL-6 releases, and coadministration of GP reversed the increase of M1 markers and enhanced the levels of M2 markers, including arginase-1 (Arg-1) expression, IL-10, brain-derived neurotrophic factor (BDNF), and glial cell-derived neurotrophic factor (GDNF) releases in the Aβ-treated microglial cells. SOCS1-siRNA, however, significantly abolished the GP-induced effects on the levels of microglial M1 and M2 markers. These findings indicated that GP attenuates Aβ-induced microglial activation by ameliorating M1/M2 states, and the process may be mediated by SOCS1.

Sign in / Sign up

Export Citation Format

Share Document