EAE （experimental autoimmune encephalomyelitis） is an established, inducible animal model employed in the study of MS （multiple sclerosis） characterized by inflammation, BBB （blood brain barrier） malfunction, de...EAE （experimental autoimmune encephalomyelitis） is an established, inducible animal model employed in the study of MS （multiple sclerosis） characterized by inflammation, BBB （blood brain barrier） malfunction, demyelination and neuronal disruption. CRF （corticotropin releasing factor） is a neuropeptide critically associated with immune function, BBB permeability, and the hypothalamic-pituitary-adrenal axis. Potential CRF targets in the brain include astrocytes, as well as endothelial cells of cerebral microvessels, since they have been reported to express CRFR （CRF receptors）. Further, both of these cell types function critically in regulating BBB permeability. CRF-BP （CRF binding protein） is also expressed in both neurons and glial cells. Changes in the cortical CRF system could be a contributing factor to the BBB disruption associated with MS/EAE and has been suggested to play a protective role against cytokine-induced inflammation. The current study assessed alterations associated with the C57BL/6 mouse model of EAE in the cortical CRF system and correlated these events with changes to the microvascular unit. Immunohistochemical confocal microscopy was used to analyze the distribution of CRF, CRF-BP, and CRFR in the mouse cerebral cortex. The authors observed a reduction in detectable CRF immunofluorescence in the EAE motor cortex, an increase in CRFBP immunoreactivity in EAE astrocytes and a concurrent reduction in astrocytic CRFR immunofluorescence. Staining techniques were used to visualize astrocytes/microvessels to document alterations in BBB integrity. Changes in the CRF system were associated with a modification of the blood brain barrier as manifested by a poorly defined astrocytic barrier in EAE microvessels. Evidence suggests that manipulation of CRF signaling pathways offers an intriguing target for interventional therapies designed to modify BBB permeability that may be beneficial for treating disease states such as MS.展开更多
OBJECTIVE: To explore the effect of Tongluojiunao injection(TLJN) prepared with Sanqi(Radix Notoginseng) and Zhizi(Fructus Gardeniae) on the interaction between brain microvascular endothelial cells(BMECs) and astrocy...OBJECTIVE: To explore the effect of Tongluojiunao injection(TLJN) prepared with Sanqi(Radix Notoginseng) and Zhizi(Fructus Gardeniae) on the interaction between brain microvascular endothelial cells(BMECs) and astrocytes in an in vitro ischemic model.METHODS: First, an in vitro model of cerebral ischemia in BMECs or astrocytes was established by oxygen-glucose deprivation(OGD). TLJN was used as a medicine of intervention. The OGD-injuredBMECs were cultured in various astrocyte-conditioned media. Cell activity, alkaline phosphatase(AKP) and γ-glutamyl transpeptidase(γ-GT) activity,interleukin-1 beta(IL-1β), and tumor necrosis factor alpha(TNF-α) content in BMECs were determined.Additionally, OGD-injured astrocytes were cultured in various BMEC-conditioned media. Cell activity, as well as expression of brain-derived neurotrophic factor(BDNF) and glial cell-derived neurotrophic factor(GDNF) in astrocytes, were detected.RESULTS: The results of paracrine signaling of normal BMECs or astrocytes showed a protective effect on each other: conditioned media from normal astrocytes improved cell viability, AKP, and γ-GT activity, and reduced IL-1β and TNF-α content of injured BMECs; conditioned media from normal BMECs improved cell viability and expression of BDNF and GDNF in injured astrocytes. However, once the BMECs or astrocytes were injured by OGD, the protective effect decreased or disappeared. The above-mentioned protective induction was effectively recovered by TLJN intervention.CONCLUSION: The therapeutic benefit of TLJN was achieved by recovering two-way induction between BMECs and astrocytes, enhancing activity of injured BMECs and astrocytes, stabilizing enzymatic barriers, promoting expression of neurotrophic factors, and inhibiting inflammatory cytokines.展开更多
文摘EAE （experimental autoimmune encephalomyelitis） is an established, inducible animal model employed in the study of MS （multiple sclerosis） characterized by inflammation, BBB （blood brain barrier） malfunction, demyelination and neuronal disruption. CRF （corticotropin releasing factor） is a neuropeptide critically associated with immune function, BBB permeability, and the hypothalamic-pituitary-adrenal axis. Potential CRF targets in the brain include astrocytes, as well as endothelial cells of cerebral microvessels, since they have been reported to express CRFR （CRF receptors）. Further, both of these cell types function critically in regulating BBB permeability. CRF-BP （CRF binding protein） is also expressed in both neurons and glial cells. Changes in the cortical CRF system could be a contributing factor to the BBB disruption associated with MS/EAE and has been suggested to play a protective role against cytokine-induced inflammation. The current study assessed alterations associated with the C57BL/6 mouse model of EAE in the cortical CRF system and correlated these events with changes to the microvascular unit. Immunohistochemical confocal microscopy was used to analyze the distribution of CRF, CRF-BP, and CRFR in the mouse cerebral cortex. The authors observed a reduction in detectable CRF immunofluorescence in the EAE motor cortex, an increase in CRFBP immunoreactivity in EAE astrocytes and a concurrent reduction in astrocytic CRFR immunofluorescence. Staining techniques were used to visualize astrocytes/microvessels to document alterations in BBB integrity. Changes in the CRF system were associated with a modification of the blood brain barrier as manifested by a poorly defined astrocytic barrier in EAE microvessels. Evidence suggests that manipulation of CRF signaling pathways offers an intriguing target for interventional therapies designed to modify BBB permeability that may be beneficial for treating disease states such as MS.
基金the National Natural Science Foundation of China(No.81273885):the Vascular-protecting Molecular Mechanism of Composition Compatibility in Gardenia and Panax Notoginseng Could be Explained by Integration ofCell Signaling Pathway NetworkCollaborative Innovation Project of the Beijing University of Chinese Medicine:"Nautical Traditional Chinese Medicine"Collaborative Innovation Center(No.522/0100604299)
文摘OBJECTIVE: To explore the effect of Tongluojiunao injection(TLJN) prepared with Sanqi(Radix Notoginseng) and Zhizi(Fructus Gardeniae) on the interaction between brain microvascular endothelial cells(BMECs) and astrocytes in an in vitro ischemic model.METHODS: First, an in vitro model of cerebral ischemia in BMECs or astrocytes was established by oxygen-glucose deprivation(OGD). TLJN was used as a medicine of intervention. The OGD-injuredBMECs were cultured in various astrocyte-conditioned media. Cell activity, alkaline phosphatase(AKP) and γ-glutamyl transpeptidase(γ-GT) activity,interleukin-1 beta(IL-1β), and tumor necrosis factor alpha(TNF-α) content in BMECs were determined.Additionally, OGD-injured astrocytes were cultured in various BMEC-conditioned media. Cell activity, as well as expression of brain-derived neurotrophic factor(BDNF) and glial cell-derived neurotrophic factor(GDNF) in astrocytes, were detected.RESULTS: The results of paracrine signaling of normal BMECs or astrocytes showed a protective effect on each other: conditioned media from normal astrocytes improved cell viability, AKP, and γ-GT activity, and reduced IL-1β and TNF-α content of injured BMECs; conditioned media from normal BMECs improved cell viability and expression of BDNF and GDNF in injured astrocytes. However, once the BMECs or astrocytes were injured by OGD, the protective effect decreased or disappeared. The above-mentioned protective induction was effectively recovered by TLJN intervention.CONCLUSION: The therapeutic benefit of TLJN was achieved by recovering two-way induction between BMECs and astrocytes, enhancing activity of injured BMECs and astrocytes, stabilizing enzymatic barriers, promoting expression of neurotrophic factors, and inhibiting inflammatory cytokines.
