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Objective To explore the effect of SFI in radiation-induced mice brain injury after 20 Gy cranial radiation.Methods The mice were divided into three groups:(1) control group,(2) RT-only group:the whole brain was irradiated with a dose of 20 Gy,(3) RT and SFI group:SFI at 20 ml/kg/d from 4 weeks after 20 Gy cranial radiation theraty(CRT).Results Morris water maze test showed that the latency of the irradiated group was longer than control group and SFI improved the cognitive function of mice (t =6.34,6.70,P ＜0.05).The expression of TNF-α reached to the highest level at 3 h after irradiation,and then it decreased but got the second higher level again at 4 weeks after irradiation.The expression of IL-1 β reached to the highest level at 72 h after irradiation and decreased until 4 weeks after irradiation.SFI decreased both expressions of TNF-α (t =11.34,9.70,6.07,P ＜ 0.05) and IL-1 β (t =12.27,5.70,7.52,P ＜ 0.05).Immune florescence staining showed that SFI reduced the number of activated microglia (t =12.35,8.64,7.82,P ＜ 0.05)and inhibited the translocation of p65 of microglia after irradiation.Conclusions Findings suggest that SFI may decrease microglial activation and suppress the expression of TNF-α and IL-1β by inhibiting the translocation of NF-κB p65 and then attenuate irradiation-induced brain injury.
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Vascular endothelial growth factor 165 (VEGF(165))-mediated autocrine stimulation of tumor cells enhances the progression to a malignant phenotype. VEGF(165)b competes with VEGF(165) and binds to vascular endothelial growth factor receptor (VEGFR), resulting in inhibition of downstream signal transduction pathways. This study was designed to investigate the role of VEGF(165)b in the migration and invasion of human lung adenocarcinoma A549 cells. The full-length of VEGF(165)b was constructed and cloned into an expression plasmid (pVEGF(165)b), and then transfected into A549 cells. Dimethylthiazolyl- 1 -2, 5-diphenyltetrazolium bromide (MTT) assay was used to detect the effect of VEGF(165)b on proliferation of transfected cells. Reverse transcription polymerase chain reaction (RT-PCR) was employed to examine the effect of VEGF(165)b on the expression of VEGF(165) in transfected cells. Wound-healing assays were used to investigate the effect of VEGF(165)b on migration of transfected cells. Matrix metalloproteinase (MMPs) activity assay and in vitro invasion assay were used to determine the role of VEGF(165)b in invasion of transfected cells. There was no significant change in proliferation of A549 cells after transfection of pVEGF(165)b, but the expression of VEGF(165), migration and invasion in A549 cells were inhibited. Furthermore, exogenous VEGF(165)b inhibited the activity of MMP9 in the supernatant of A549 cells and the subsequent invasion capacity of those cells. We therefore conclude that exogenous VEGF(165)b can inhibit the expression of VEGF(165), as well as the migration and invasion of A549 cells, but has no effect on the proliferation of A549 cells.
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Objective To explore the inhibitory effects of Tanshinone Ⅱ A on the radiationinduced microglia activation and the possible mechanism.Methods Microglia cells BV-2 were irradiated with 2,4,8,16,and 32 Gy doses or sham-irradiated in presence or absence of 1.0 μg/ml Tanshinone Ⅱ A for 12 h,respectively.The effects of Tanshinone Ⅱ A on radiation-induced pro-inflammatory cytokines were evaluated using real-time PCR.The expression level of NF-κB p65 in cytoplasm and nucleus was measured by using Western blot.Immunofluorescence staining and confocal microscopy analysis were applied to detect the expression of γ-H2AX and p65 post-irradiation.Results The microglia cells were activated at 16,32 Gy post-irradiation.Radiation-induced release of the pro-inflammatory cytokines in BV-2 cells was detectable after irradiation.Tanshinone Ⅱ A decreased radiation-induced release of proinflammatory cytokines(t=5.56,P ＜ 0.05).Furthermore,western blotting showed that Tanshinone Ⅱ A could attenuate the nuclear translocation of NF-κB p65 submit post-irradiation.Immunofluorescence staining showed that γ-H2AX foci formation while p65 translocation into nucleus post-irradiation.Conclusions Tanshinone Ⅱ A exerts anti-inflammatory properties by suppressing the transcription of proinflammatory cytokine genes that might be associated with NF-κB signaling pathway.It is postulated that irradiation causes immediate cellular reaction and DSB triggers the molecular response which leads to NFκB pathway activation.
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Objective To explore the inhibitory effects of Corilagin on the production of proinflammatory cytokines in microglia induced by radiation. Methods The cytotoxicity of Corilagin was measured by MTT assay. Microglia BV-2 cells were irradiated 0 or 32 Gy after pretreated with Corilagin for 12 hours. Realtime-PCR was used to detect the mRNA levels of inflammatory cytokines, such as IL-1β,TNF-α on several time-points. The content of nitric oxide (NO) was determined with nitrate reductase method. The translocation of NF-κB was measured by Western blot and immunocytochemical stain.Confocal microscopy was used to observe the expression of Iba-1 and Nemo. Results No cytotoxicity was detected on BV-2 cells with 1-10 μg/ml Corilagin. Iba-1 expression in microglia cells was activated by irradiation, the expression levels of inflammatory cytokines, such as IL-1β, TNF-α and NO were also elevated. Whereas, the production of IL-1 β, TNF-α in activated microglia cells was significantly inhibited with 5 μg/mL corilagin ( tIL-1β = 6. 341, tTNF-α = 3.41 1, tNO = 3. 134, P ＜ 0. 05 ). Corilagin significantly inhibited the expression of Nemo and the translocation of NF-κB p65. Conclusion Corilagin could inhibit the activation of irradiated microglia cells and down-regulate the expression of inflammatory cytokines, via inhibition of the NF-κB signaling pathway.