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Objective:To investigate any effect of repeated transcranial magnetic stimulation (rTMS) on the expression of P2X7 receptor (P2X7R) and glial fibrillary acid protein (GFAP) in the prefrontal cortex and hippocampus of mice modeling depression.Methods:Thirty C57BL/6 mice were divided into a control group ( n=10) and a depression group ( n=20). The mice of the control group were raised in group (five mice per cage), while those of the depression group were kept alone for six weeks to induce depression. Among them, 16 were successfully modeled and randomly divided into a model group ( n=8) and an rTMS group ( n=8). The rTMS group received five sessions per week of 10Hz rTMS for 4 weeks. Any changes in depression-like behavior were observed and the expression of P2X7R and GFAP in the prefrontal cortex and hippocampus was measured. Results:Compared to the control group, a significant decrease was observed in the sucrose consumption rate in the sucrose preference test, in the distance moved in the open field test and in the expression of GFAP protein. But there was a significant increase in the immobile time in the tail suspension test and in the expression of P2X7R protein in the prefrontal cortex and hippocampus in the model group. At the conclusion of the experiment the differences in the sucrose consumption rate, the distance moved, GFAP protein expression, immobile time and P2X7R protein expression between the rTMS and the model group were all statistically significant.Conclusion:rTMS can reduce depression-like behavior, at least in mice. That may be related to inhibiting P2X7R expression and promoting GFAP expression in the prefrontal cortex and hippocampus.
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Objective:To investigate the inhibitory effect of specific inhibitor of necroptosis necrostatin-1 (Nec-1) on necroptosis of retinal ganglion cells (RGCs) in rats with acute ocular hypertension.Methods:Twenty-four adult male Sprague Dawley rats were randomly divided into normal control group, model control group, Nec-1 treatment group and negative control group by random number table method, with 6 rats in each group.High intraocular pressure (IOP)-induced ischemia and reperfusion model was established through anterior chamber irrigation of 0.9% sodium chloride solution in left eyes of the rats, raising the IOP to 110 mmHg (1 mmHg=0.133 kPa) for 60 minutes.Nec-1 (4 mmol/L, 2 μl) or dimethyl sulfoxide (2 μl) was intravitreally injected immediately in Nec-1 treatment group and negative control group following modeling, respectively, according to grouping.No intervention was administered to the normal control group.Paraffin sections of rat retinas of the left eyes in different groups were prepared seven days after modeling.The retinal structure was observed by hematoxylin-eosin staining, and the expression levels of thymocyte antigen-1 (Thy-1) and glial fibrillary acidic protein (GFAP) were detected via immunohistochemical staining.All animal experiments were approved by an Ethics Committee of Tianjin Union Medical Center (No.2017 Quick audit C01).Results:Seven days after modeling, compared with normal control group, the retinal nerve fiber layer was thinner in model control group and negative control group, and the RGCs were arranged loosely, and cells in the inner nuclear layer were reduced and arranged disorderly, and cells in the outer nuclear layer were normal or enlarged.Compared with model control group and negative control group, the nerve fiber layer was thickened and the number of RGCs was significantly increased in Nec-1 treatment group.The number of Thy-1-positive RGCs was decreased in model control group, negative control group and Nec-1 treatment group than normal control group, and there were more Thy-1-positive RGCs in Nec-1 treatment group than model control group and negative control group.The integrated absorbance ( A) value of GFAP protein in normal control group, model control group, negative control group and Nec-1 treatment group was 47.209±15.311, 116.220±18.194, 116.382±19.020, 92.818±10.236, respectively, showing statistically significant differences among them ( F=24.675, P<0.001). The integrated A value of GFAP protein was significantly increased in model control group, negative control group and Nec-1 treatment group than normal control group, and the integrated A value of GFAP protein in Nec-1 treatment group was lower than that in model control group and negative control group, with statistically significant differences (all at P<0.05). Conclusions:Nec-1 can promote RGCs survival by inhibiting the necroptosis of RGCs in rats with acute intraocular hypertension.
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Objective To study effect of electroacupunture on reactive astrogliosis of rats after spinal cord injury. Methods A total of 36 SD rats were randomly divided into the sham group, model group and electroacupuncture group, 12 rats in each group. The sham group only underwent laminectomy without spinal cord injury, and the impactor was used to establish spinal cord injury model in model group and electroacupuncture group. The electroacupuncture group was given electroacupuncture after operation, while the rest groups were given the same condition of grabing. The Basso Beattie Bresnahan (BBB) scores of rats were recorded daily after operation and rats were sacrificed on the 14th day after administration. Nissl staining was used to observe the morphology of neurons. The expression of GFAP was detected by immunofluorescence. The expression of GFAP protein was detected by Western blot. Results The fifth days after the intervention, the BBB score of the electroacupuncture group was significantly higher than that of the model group (P<0.05). Compared with the model group, the expression of GFAP in electroacupuncture group significantly decreased (F=23.831, P<0.001), the expression of GFAP protein in electroacupuncture group significantly decreased (F=15.883, P<0.001), the expression of IL-1β and TNF-α in electroacupuncture group significantly decreased (F=82.773, 113.487, P<0.001). Conclusions The electroacupuncture can inhibit the expression GFAP and astrogliosis to inhibit inflamation after spinal cord injury in rats, which is conducive to the repair of neurons and the recovery of limb motor function in rats.
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This paper was aimed to investigate the effects of emodin on oxidative stress and inflammatory response in rats with acute spinal cord injury（SCI）, and to explore the protective mechanism of emodin on neurons after SCI. Rat SCI models were established using a modified Allen's method. One hundred and ninety five Sprague-Dawley rats were randomly divded into sham (group A), model (group B), emodin group of 20 mg·kg⁻¹(group C), emodin group of 40 mg·kg⁻¹(group D), emodin group of 80 mg·kg⁻¹(group E). Functional recovery was evaluated using the Basso-Beattie-Bresnahan (BBB) scale and inclined plate test on the 3rd, 7th, 14th and 28th day. On the 7th day after SCI, neuron nylon body was observed with toluidine blue staining. The changes of myelinated nerve fibers were observed by transmission electron microscope. Nrf2, HO-1, NQO1, GFAP, NF-κB protein were detected by Western blot. The content of TNF-α, IL-1β, IL-6 were detected by ELISA. Immunofluorescence staining was performed to observe the expression of the GFAP, NG2 and ED-1. The BBB and inclined plate scores of group C, D and E were higher than group B on the 7th, 14th, 28th day,the difference was statistically significant (<0.05). On the 7th day, Nylon Body of group B and C started to fuse,the fusion of group D and E were significantly alleviated than group B and C. Transmission electron microscope showed that the changes of demyelination were obvious in group B and C, group D and E were significantly improved than group B and C. Western blot showed that Nrf2, HO-1, GFAP, NF-κB protein expression,group C, D, E compared with group B, NQO1 protein expression, group D, E compared with group B, the difference was statistically significant (<0.05). ELISA showed that the content of TNF-α, IL-6,group C, D, E compared with group B,the content of IL-1β,group D, E compared with group B, the difference was statistically significant (<0.05)；Immunofluorescence showed that the expressions of GFAP and NG2 in group C, D and E were higher than that in group B, and the group D and E were more obvious. The expression of ED-1 in group C, D, E were decreased significantly compared with group B. Emodin has protective effect on neurons after SCI. The mechanism may connect with activting Nrf2-ARE pathway, reducing the expression of NF-κB, ED-1, TNF-α, IL-1β, IL-6, and promoting the expression of GFAP and NG2.
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Objective To explore the effects of the new melatonin nonselective agonists Neu-P11 on intraocular pressure (IOP) and glial fibrillary acid protein (GFAP) expression in the retina of acute high IOP rat.Methods Twenty-four male Sprague-Dawley rats were randomly divided into 4 groups (6 cases in each group):Normal IOP with local treatment (NIL) group,high IOP with local treatment (HIL) group,HILwith melatonin treatment (HIL-M) group,HIL with Neu-P11 treatment (HIL-N) group.10 μL normal saline was instilled in NIL group and HIL group,while 10 μL 100 μmol · L-1 Mel/Neu-P11 treated in HIL-M group and HIL-N group.After 2 hours of rest,rats were placed in the Trendelenburg position duration 45 minutes.And then,IOP was measured every hour for 6 hours,and repeated it for a week.The excessive sodium pentobarbital was injected to SD rats at the end of the experiment.The rat eyeballs were took out to perform HE and immunohistochemical staining to detect retina GFAP protein expression.Results After a week,IOP in HIL group was (41.26 ± 1.73) mmHg (1 kPa =7.5 mmHg),NIL group was (13.61 ± 0.55) mmHg,which mean the Trendelenburg could induce high IOP in SD rats.Compared with the NIL group,the retinal becoming thick,the level of organization was not clear and the expression of GFAP protein was quite high in HIL group.At the same time,the GFAP protein expression and IOP were significantly weakened in HIL-M group and HIL-N group compared with HIL group.Conclusion Neu-P1 1 can reduce IOP,inhibit the activation of gliocyte,and decrease the expression of GFAP to protect the retina.
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Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.
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Objective To explore the influence of tetrahydroxystilbene glucoside ( TSG ) on the neuron proliferation in epileptic rats and the related mechanisms.Methods 54 healthy SD male rats were randomly divided into three groups: sham-operated group, epilepsy group and epilepsy with TSG treatment group.The epilepsy group was established by stereotactic brain trace injection with kainic acid ( KA ) . TSG solution ( 3 mg/kg ) was injected intraperitoneally at 6 hours after the epilepsy group established, and then q.d.for consecutive 42 days.The sham-operated group and epilepsy group were injected with normal saline.The influence of TSG on cell proliferation of rat hippocampal dentate gyrus BrdU-positive granular cells was observed by immunohistochemistry.Results Compare with the epilepsy group, the amount of glial fibrillary acid protein ( GFAP)-positive astrocytes was significantly reduced and dentate gyrus BrdU-positive cells were significantly increased in the TSG group ( P <0.01 ) .Conclusions Tetrahydroxystilbene glucoside ( TSG) promotes neurogenesis of neural stem cells and neurons, and inhibits the growth of hippocampal dentate gyrus astrocytes in epilepsy rats.
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Background Our previous study showed that the expression level of glial fibrillary acid protein (GFAP) increases in astrocytes and Müller cells of retina in chronic hypertensive eye,and this change was clarified to be associated with the damage process of the retinal ganglion cells (RGCs).Aquaporin 4 (AQP4) exists in neural glial cells,so we conjecture AQP4 plays a role in the regulating GFAP expression in glaucomatous eye.Objectives This study was to investigate whether AQP4 gene can regulate the expression of GFAP in retina and explore the effect of AQP4 on RGCs damage of glaucoma.Methods Chronic ocular hypertensive eye models were established by cauterizing the scleral venous in the left eyes of 30 male AQP4-/-mice and 30 male wild type (WT) mice with the same background,and the right eyes served as control eyes.The intraocular pressure (IOP) was measured with Icare rebound tonometer at 1 day,3,7,14 and 28 days respectively and the retinas were isolated from 6 of each types of mice at the corresponding time points.The expression of GFAP in the retina was detected by Western blot.The use and care of the experimental animals followed ARVO Statement.Results The IOP was significantly higher in the model eyes than that of the control eyes 1 day,3,7,14 and 28 days in both AQP-/-mice (t =15.29,16.02,13.77,14.34,12.40,all at P＜0.05) and WT mice (t =17.65,14.91,15.97,13.41,12.53,all at P ＜0.05).GFAP was expressed in the control eyes both of the AQP4-/-mice and the WT mice.The expressing level of GFAP (GFAP/β-actin) in retinas was 1.00±0.00,1.99±0.29,4.05±0.69,4.47±0.48,3.21±0.35 and 3.25±0.53 in the control eyes and 1-,3-,7-,14-,28-day model eyes of WT mice; and those in the AQP4-/-mice were 1.00±0.00,1.69±0.31,2.27 ±0.55,2.79 ± 0.39,1.93 ± 0.31 and 1.54 ± 0.40,with a significant difference in the expressing level of GFAP in various time points (F =9.54,P＜0.05).In addition,significant gradually elevation of GFAP expression were seen in the WT mice and gradually decline of GFAP expression was found in the AQP4-/-mice with the lapse of time (all at P＜0.05).No significant difference was seen in the expression of GFAP in the control eyes between the WT mice and AQP4-/-mice (P＞0.05).However,the expression level of GFAP in retina was significantly higher in the WT mice than that of A QP4-/-mice 3,7,14 and 28 days after operation (t =4.51,7.95,6.12,5.76,all at P＜0.01).tonelusions In chronic high IOP mice,AQP4 gene plays an important role in retinal damage by upregulating the expression of GFAP in retina and promoting the activation of RGCs.AQP4 probably is a new target of treatment of glaucoma.