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1.
AIM: To explore the time-dependent change of Ski protein expression in normal and activated astrocytes in rats.METHODS: The astrocytes were obtained from rat cerebral cortex and cultured in vitro. The astrocytes were treated with LPS and scratch injury for activation. Western blot analysis was used to determine glial fibrillary acidic protein (GFAP) and Ski protein levels in activated astrocytes at a series of time points. The indirect immunofluorescence staining method was performed to detect the location of Ski protein in the astrocytes.RESULTS: The protein of GFAP was naturally expressed in the astrocytes, beginning to increase after treated with LPS and scratch injury. Little protein expression of Ski in the normal astrocytes was observed. The Ski protein expression began to increase after treated with 1 mg/L LPS, peaked at 4 d (P<0.05) and then deceased, but was stills higher than that in the normal cells. The protein expression level of Ski after scratch injury was highly consistent with above mentioned. Ski was mainly observed in the nucleus of the normal cells and the cells treated with LPS for 6 d, while it was observed in the cytoplasm 2 and 4 d after treated with LPS.CONCLUSION: The protein of Ski is expressed in the astrocytes, and the expression level is increased in activated astrocytes, mainly located in the nucelus. Ski may plays an essential roles in the processes of activation and proliferation of astrocytes.  相似文献   

2.
AIM: The present study was designed to examine the changes in glial fibrillary acidic protein (GFAP) expression during cerebral ischemia and the effects of ginkgolide B on GFAP expression. METHODS: The focal thrombotic cerebral ischemia was formed by photochemistry-induced in tree shrews. GFAP stained by ABC immunohistochemistry and absorbance were measured with image analyze system. RESULTS: GFAP expression in astrocytes increased significantly (P<0.01) at 24 h and kept in higher level at 72 h (P<0.01) within penumbra after focal cerebral ischemia. GFAP expression declined when the animals were given GB at 6 h after thrombotic cerebral ischemia. CONCLUSIONS: Neuronal necrosis resulted in GFAP expression in astrocytes after local cerebral ischemia and GB protected neurons by antagonizing PAF receptor and inhibiting GFAP expression.  相似文献   

3.
AIM: To observe the inhibitory effect of madecassoside on the LPS-stimulated microglia and to investigate its possible mechanism. METHODS: Microglia cells of neonatal Sprague-Dawley (SD) rats were cultured, isolated and purified. Microglia cells were activated with lipopolysaccharide (LPS). The inhibitory effect of madecassoside on microglia was measured by MTT assay. Tumor necrosis factor alpha (TNF-α), interleukin 1β (IL-1β) were detected by ELISA. Cell cycle and apoptotic rate were evaluated by flow cytometry. The expression of TLR4 was detected by Western blotting. The expression of NF-κB was detected by RT-PCR. RESULTS: LPS induced the proliferation of microglia and release inflammatory cytokines significantly. Compared with LPS group, madecassoside inhibited the proliferation of microglia induced by LPS in a dose dependent manner. The IC50 value of madecassoside was 10.97 nmol/L to microglia after incubation for 48 h. Madecassoside also decreased the levels of TNF-α and IL-6, increased the ratios of microglia at the G2 phase and the apoptotic rate, decreased the expression of TLR4 and NF-κB significantly (P<0.05). CONCLUSION: Madecassoside has inhibitory effects on the proliferation of LPS-stimulated microglia, by which the mechanism may be related to inhibition of the expression of TLR4 and NF-κB, change of cell cycle distribution and induction of microglia apoptosis.  相似文献   

4.
AIM: To investigate the inhibitory effect of saikosaponin a (SSa) on pentylenetetrazole (PTZ)-induced activation of hippocampal astrocytes in mice. METHODS: Hippocampal astrocytes were isolated and cultured. The cells were randomly divided into control group, PTZ group, PTZ+0.625 mg/L SSa group and PTZ+1.25 mg/L SSa group. The cells were identified by detection of glial fibrillary acidic protein (GFAP). The cell viability was measured by MTT assay. The cell cycle was analyzed by flow cytometry. The expression of GFAP and connexin 43 (Cx43) was mea-sured by ELISA. The level of apoptosis was determined by flow cytometry and Hoechst 33258 staining. RESULTS: The primary hippocampal astrocytes grew by adherent culture, and the processes of the astrocytes were obvious. Immunofluorescence showed positive GFAP expression in the astrocytes. Compared with control group, the viability of the cells and the percentage of the cells in G2/M phase in PTZ group were significantly increased (P<0.05), and the expression of GFAP and Cx43 was also markedly increased (P<0.05). Compared with PTZ group, the viability of the cells and the percentage of the cells in G2/M phase were obviously decreased in PTZ+0.625 mg/L SSa group and PTZ+1.25 mg/L SSa group, and the expression of GFAP and Cx43 was also reduced, whereas the percentage of apoptotic cells was significantly increased (P<0.05).CONCLUSION: SSa significantly suppresses PTZ-induced activation of hippocampal astrocytes, inhibits the cell proliferation and induced apoptosis.  相似文献   

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6.
AIM: To observe the effect of platelet-activating factor (PAF) on cultured neuronal viability and glial fibrillary acidic protein (GFAP) expression in cultured astrocytes. METHODS: Neurons and astrocytes obtained from the brain cortex of the embryo and newborn mice respectively were cultured and purified, and they were divided into the control and experimental groups. PAF was added into the experimental groups at concentrations of 4, 8 and 16 μmol/L. Each group was cultured for 4 h, 24 h and 72 h, respectively. MTT method and immunohistochemistry were used to observe the neuronal viability and GFAP expression in astrocytes, respectively. RESULTS: During different time after adding PAF at different concentrations into cultured neurons and astrocytes, respectively, neuronal viability declined, and the number of astrocytes decreased, but GFAP expression in survival astrocytes increased. The effects were shown to be in a concentration-dependent manner. CONCLUSION: PAF decreases the neuronal viability directly and influences the neuronal survival indirectly by astrocytes.  相似文献   

7.
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8.
AIM: To investigate the effects of propofol on the expression of apoptosis-inducing factor (AIF) and cell apoptosis in brain tissues of rats with lipopolysaccharide(LPS)-induced brain injury. METHODS: Seventy-two male and female SD rats weighing 220~250 g were randomly divided into 3 groups (n=24 each). Cerebral edema was induced by injection of LPS at 1 mg/kg via left internal carotid artery in LPS group and LPS+propofol group. In control group, equal volume of normal saline was administered instead of LPS. The rats in LPS+propofol group received intraperitoneal injection of propofol at 100 mg/kg immediately after LPS administration. Six rats in each group were decapitated 6 h, 12 h, 24 h or 48 h after operation and the frontal lobe cortex were immediately removed for determination of the water content. The apoptotic neurons were detected by Annexin V-PI staining. The protein levels of AIF, NF-κB and caspase-3 were measured by immunohistochemistry. The protein expression of AIF was detected by Western blotting analysis. RESULTS: Compared with control group, the brain water content, the number of neuronal apoptosis and the protein expression levels of AIF, NF-κB and caspase-3 were significantly increased in LPS group and LPS+propofol group. Compared with LPS group, the results mentioned above were markedly reduced in LPS+propofol group. CONCLUSION: Propofol attenuates LPS-induced brain injury by decreasing AIF protein expression and inhibiting apoptosis.  相似文献   

9.
AIM: To explore the expression of CD14 in rat Kupffer cells (KCs). METHODS: In rat KCs induced by LPS or the mediators from KCs induced by LPS, the changes of CD14 expression were measured by RT-PCR and immunohistochemistry.The expressions of TNFα mRNA、IL-6 mRNA or the concentrations of TNFα、IL-6 were estimated by in situ hybridization and radioimmunoassay, respectively. RESULTS: LPS increased the expression of CD14 in KCs in a dose-dependent fashion (LPS, 1 μg/L-10 mg/L) and in a time-dependent fashion(0.5 h-24 h, peaked at 3-6 hours). While the expression of CD14 in KCs stimulated by the active mediators from KCs which had been exposed to LPS 1 hour were obviously increased. CONCLUSIONS: There was a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14. It is implied that the increase in CD14 expression may be induced by LPS and the cytokines produced by KCs, it also reveals that there is a auto-regulated loop in CD14 expression.  相似文献   

10.
AIM:To observe the activity and distribution of astrocytes and glial fibrillary acid protein(GFAP) after middle cerebral artery occlusion (MCAO). METHODS:The rat MCAO model was made by two-kidney, tow clip renovascular hypertensive rat stroke prone(RHRSP). Rats were killed and brain samples were collected at the end of 1,3,6 and 9 weeks after MCAO, respectively. The ultrastructure of astrocytes was determined at broder of infarct (A area); distant of infarct (B area) and opposite of hemisphere (C area) by electron microscope. The number and optical density of GFAP-positive cells were also observed. RESULTS:The astrocyte proliferation distributed in the whole brain after MCAO. The highest numbers of GFAP-positive cells were observed at A area, then B area. The lowest numbers of GFAP positive cells were found in C area. The time course of GFAP-positive cell change was that the highest number was observed at 1 week after MCAO, then decreased by time from 3, 6 weeks to 9 weeks. The optical density of GFAP-positive cells showed the same patterns. CONCLUSION:The correlation between astrocyte proliferation and tissue damage after MCAO can be estimated by GFAP expression. The astrocyte proliferation plays an important role in healing process after MCAO.  相似文献   

11.
AIM: To investigate the activity of astrocytes and autophagy-related changes after radiation-induced brain injury (RBI) in rats.METHODS: A total of 36 Sprague-Dawley rats, weighing 180~200 g, were trained for 4 d in the Morris water maze. They were randomly divided into sham group, model group and 3-methyladenine (3-MA) group. The rats in model group and 3-MA group were given single whole-brain X-ray irradiation at a dose of 20 Gy after intraperitoneal anesthesia. After the irradiation was completed, the rats in model group was given 5 μL of NaCl into the lateral ventricle, and the rats in 3-MA group was injected with 3-MA at 600 nmol into the lateral ventricle. After 8 weeks of feeding, Morris water maze was used for measuring the learning and memory abilities. The brain tissues were taken and HE staining was used to observe the pathological changes of the hippocampus. The protein level of GFAP was determined by immunohistochemistry and Western blot for evaluating astrocyte activity. Dual fluorescence staining of GFAP and LC3 was performed for evaluating the changes of autophagy in the astrocytes. The protein level of cleaved caspase-3 detected by Western blot and TUNEL staining in the ipsilateral hippocampus were used to evaluate the apoptosis. The contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were examined by ELISA to assess the inflammatory response in the hippocampus.RESULTS: Radiation inhibited astrocyte activity, activated autophagy in astrocytes, and aggravated brain damage. 3-MA promoted the activation of astrocytes and promoted the repair of brain tissue damage.CONCLUSION: The injury of rat hippocampus after radiation is obvious, and the number of astrocytes is significantly reduced. 3-MA significantly attenuates the damage. This finding may provide a new approach for the treatment of radiation-induced brain injury.  相似文献   

12.
AIM: To investigate the effects of dexmedetomidine on astrocytes in rats with focal cerebral ischemia-reperfusion. METHODS: Sixty female SD rats, weighing 230~250 g, were randomly divided into sham operation group, ischemia-reperfusion group, dexmedetomidine preconditioning group 1 and dexmedetomidine preconditioning group 2. The model of middle cerebral artery occlusion (MCAO) was established by thread embolism of middle cerebral artery. In sham operation group, the carotid arteries were exposed without performing MCAO. In ischemia-reperfusion group, NS was injected intraperitoneally 30 min before focal cerebral ischemia-reperfusion. The rats in dexmedetomidine preconditioning group 1 and dexmedetomidine preconditioning group 2 received intraperitoneal injection of dexmedetomidine at doses of 20 μg/kg and 40 μg/kg, respectively. The neurological scores were studied, and the pathological changes were observed under microscope with HE staining. The expression of glial fibrillary acidic protein (GFAP) and tumor necrosis factor α (TNF-α) in astrocytes was detected by the methods of immunohistochemistry and immunoblotting 24 h after cerebral ischemia-reperfusion. RESULTS: No neurological change was observed in sham operation group. The neurological deficiency scores in ischemia-reperfusion group were markedly higher than those in dexmedetomidine preconditioning group 1 and group 2 (P<0.05). Compared with sham operation group, the expression of GFAP and TNF-α in astrocytes and the level of GFAP increased significantly 24 h after focal cerebral ischemia-reperfusion. Pretreatment with dexmedetomidine significantly attenuated the expression of GFAP and reduced the infarct size and inflammation. CONCLUSION: Dexmedetomidine has a neuroprotective effect on focal cerebral ischemia-reperfusion injury by inhibiting the astrocytes.  相似文献   

13.
AIM:To study the effect of activated protein C (APC) at different concentrations on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS).METHODS:The HUVECs were induced by LPS (1.0 mg/L) as apoptotic model that was administered by different concentration of APC (10 μg/L or 50 μg/L). Meanwhile, the control group and induced apoptosis group induced by LPS (1.0 mg/L) stimulation were also set up. The changes of cellular ultrastructures were observed under electron microscope. The DNA ladder and TUNEL fluorescent staining were measured in cells. Annexin-Ⅴ/PI double staining was used to measure the cell apoptosis rate by flow cytometry. Cell survival rate was measured by MTT assay. The proliferating cell nuclear antigen (PCNA) expression levels in cells were also measured by Western blotting to reflect the proliferation of the cells.RESULTS:There were significant apoptotic changes in the cells induced by LPS, but the apoptotic changes were reduced and apoptosis rates were decreased in the cells treated with APC. Meanwhile, cell survival rate and the protein levels of PCNA were increased after APC treatment, particularly at the concentration of 50 μg/L, which showed difference when compared with those induced apoptosis group by LPS (P<0.05).CONCLUSION:APC can inhibit HUVECs apoptosis induced by LPS and promote cell proliferation, thus protect the cells from injury.  相似文献   

14.
AIM: To investigate the differentiation-inducing effect of cell differentiation agent-2 (CDA-2) in human SWO-38 glioma cell line in vitro.METHODS: The inhibitory effect of CDA-2 on cell proliferation was assessed by MTT assay and colony formation assay.Cell morphology was determinded by light microscopy observation,and the expression of GFAP (glial fibrillary acidic protein) was detected by immunohistochemistry and Western blotting.Western blotting was also applied to explore the expression of PPARγ and COX-2.RESULTS: The data showed that CDA-2 inhibited proliferation and induced differentiation of SWO-38 cells.The inhibition efficiency was time-dependent and dose-dependent .The IC50 of CDA-2 was (2.33±0.37) g/L and (0.51±0.01) g/L,respectively when cells were treated for 72 h and 10 days.CDA-2 caused differentiation of human glioma cells as indicated by outgrowth of long processes and expression of astrocyte marker GFAP.Simultaneously,the expression of PPARγ increased after 3 h of CDA-2 treatment，while the expression of COX-2 decreased after 48 h of CDA-2 treatment.CONCLUSION: CDA-2 inhibits proliferation and induces differentiation of SWO-38 cells.These effects may be through increasing cellular GFAP,PPARγ level and decreasing COX-2 expression induced by CDA-2.  相似文献   

15.
AIM: To investigate the mechanism and the effect of glycogen synthase kinase 3β (GSK-3β) inhibitor (2’Z,3'E)-6-bromoindirubin-3'-oxime (BIO) on the protein expression of β-catenin and Bcl-2, and proliferation and apoptosis in colon carcinoma SW480 cells.METHODS: The immunohistochemical staining and Western blotting were performed to detect the protein expression of β-catenin, cyclin D1 and Bcl-2. The cell cycle distribution and apoptotic rate were detected by flow cytometry. The morphologic features of SW480 cells before and 24 h after BIO exposure at different concentrations were observed under microscope with HE staining.RESULTS: Compared with the untreated SW480 cells, the protein expression of β-catenin significantly increased and some β-catenin positive nuclear staining positive cells appeared in BIO treated cells. and The cells exposed to BIO showed that the cyclin D1 protein and the cells in S stage and G2/M stage moderately increased, the protein level of Bcl-2 moderately decreased, and the cell apoptosis rate was significantly lower than those in control cells. Furthermore, the morphological changes of the SW480 cells were observed 24 h after BIO treatment. CONCLUSION: Our results indicate that GSK-3β inhibitor BIO participates in the cellular processes of promoting proliferation and inhibiting apoptosis in colon carcinoma cells. The mechanisms are mainly associated with activating the β-catenin pathway and regulating the balance of Bcl-2 pathway, and the up-regulation of β-catenin is most likely the possible factor for SW480 cell regression.  相似文献   

16.
AIM To investigate the effects of ketamine at subanesthetic dose on α-synuclein and astrocyte in mice with Parkinson disease (PD) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). METHODS Thirty-six healthy male C57BL/6 mice were randomly divided into 3 groups (12 mice per group): NaCl group (intraperitoneal injection of saline), MPTP group (intraperitoneal injection of MPTP) and ketamine group (intraperitoneal injection of MPTP and ketamine). The behavioral differences among the mice in the 3 groups were examined by tail suspension test and gait analysis test. Immunofluorescence staining and Western blot were used to detect the expression of α-synuclein and glial fibrillary acidic protein (GFAP) in the substantia nigra (SN), caudate putamen (CP) and visual cortex (CX). RESULTS According to the results of tail suspension test and gait analysis test, the mice in MPTP group showed increased duration of immobility and shortened step length compared with NaCl group, while those in ketamine group showed decreased duration of immobility and expanded step length compared with MPTP group (P<0.05). According to the results of immunofluorescence staining and Western blot, the mice in MPTP group showed significantly increased expression of α-synuclein and GFAP in SN, CP and CX compared with NaCl group, while those in ketamine group showed significantly decreased expression of a-synuclein and GFAP in SN, CP and CX compared with MPTP group (P<0.05). CONCLUSION Ketamine at subanesthetic dose inhibits the expression of α-synuclein and the proliferation of astrocytes in MPTP-induced PD mice.  相似文献   

17.
AIM: To investigate the effects of eicosapentaenoic acid(EPA) on the expression of nuclear factor kappa B(NF-κB) and release of vascular endothelial growth factor(VEGF), IL-1α and IL-6 in cultured human umbilical vein endothelial cells(HUVECs) stimulated by lipopolysaccharide(LPS).METHODS: HUVECs were obtained from cell strain and cultured in vitro. HUVECs were divided into 4 groups: control group, LPS group, 0.030 g/L EPA treatment group and 0.050 g/L EPA treatment group. The cells were cultured with LPS alone in LPS group and incubated with EPA for 1 h in the EPA pretreatment groups at the concentrations of 0.030 g/L and 0.050 g/L before LPS stimulation. Twenty-four hours after stimulated by LPS, the protein expression of NF-κB p65 in HUVECs were assessed by Western blotting analysis at different time points. The production of VEGF, IL-1α and IL-6 in cultured HUVECs was evaluated by ELISA. The effects of EPA on the protein expression of NF-κB p65 and the production of VEGF, IL-1α and IL-6 in HUVECs challenged by LPS were also determined.RESULTS: Compared with control group, the protein expression of NF-κB p65 was significantly increased in HUVECs induced by LPS and was inhibited by EPA. Compared with control group, the protein expression of VEGF, IL-1α and IL-6 was dramatically increased in HUVECs induced by LPS and most of the increase was inhibited by EPA.CONCLUSION: LPS enhances the protein expression of NF-κB and the release of VEGF, IL-1α and IL-6. EPA inhibits the protein expression of NF-κB, and the production of VEGF and the inflammatory cytokines in cultured HUVECs stimulated by LPS, indicating that EPA may be useful for preventing and treating neovascular and inflammatory diseases.  相似文献   

18.
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19.
WANG Shi-meng 《园艺学报》2017,33(2):289-296
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20.
XU Yan  SHU Shi-yu 《园艺学报》2020,36(7):1170-1177
AIM To study the effects of HET0016 on the proliferation and migration of microglia stimulated by lipopolysaccharide (LPS). METHODS Primary microglia from neonatal SD rats were isolated, purified and cultured. CCK-8 assay was performed to detect the effect of HET0016 on the viability of microglia after treatment with LPS. The levels of 20-hydroxyeicosatetraenoic acid （20-HETE）, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured by ELISA. The proportion of S phase was evaluated by flow cytometry. The cell migration ability was detected by Transwell assay and scratch wound healing assay. The protein expression of NF-κB p50 and p65 was determined by Western blot. RESULTS LPS induced the increases in the proliferation and migration of microglia and the release of inflammatory cytokines (P<0.05). Compared with LPS group, HET0016 inhibited the cell proliferation and migration (P<0.05), decreased the levels of TNF-α and IL-1β (P<0.05), and reduced the expression of NF-κB p50 and p65 (P<0.05). CONCLUSION HET0016 has inhibitory effects on the proliferation and migration of microglia induced by LPS, and reduces the release of inflammatory cytokines. The mechanism may be related to NF-κB signaling pathway.  相似文献   

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