Changes and significance of serum ubiquitin carboxyl-terminal hydrolase L1 and glial fibrillary acidic protein in patients with glioma

BACKGROUND Brain gliomas are malignant tumors with high postoperative recurrence rates.Early prediction of prognosis using specific indicators is of great significance.AIM To assess changes in ubiquitin carboxy-terminal hydrolase L1(UCH-L1)and glial fibrillary acidic protein(GFAP)levels in patients with glioma pre-and postoperatively.METHODS Between June 2018 and June 2021,91 patients with gliomas who underwent surgery at our hospital were enrolled in the glioma group.Sixty healthy volunteers were included in the control group.Serum UCH-L1 and GFAP levels were measured in peripheral blood collected from patients with glioma before and 3 d after surgery.UCH-L1 and GFAP levels in patients with glioma with different clinicopathological characteristics were compared before and after surgery.The patients were followed-up until February 2022.Postoperative glioma recurrence was recorded to determine the serum UCH-L1 and GFAP levels,which could assist in predicting postoperative glioma recurrence.RESULTS UCH-L1 and GFAP levels in patients with glioma decreased significantly 3 d after surgery compared to those before therapy(P<0.05).However,UCH-L1 and GFAP levels in the glioma group were significantly higher than those in the control group before and after surgery(P<0.05).There were no statistically significant differences in preoperative serum UCH-L1 and GFAP levels among patients with glioma according to sex,age,pathological type,tumor location,or number of lesions(P>0.05).Serum UCH-L1 and GFAP levels were significantly lower in the patients with WHO grade I-II tumors than in those with gradeⅢ-IV tumors(P<0.05).Serum UCH-L1 and GFAP levels were lower in the patients with tumor diameter≤5 cm than in those with diameter>5 cm,in which the differences were statistically significant(P<0.05).Glioma recurred in 22 patients.The preoperative and 3-d postoperative serum UCH-L1 and GFAP levels were significantly higher in the recurrence group than these in the non-recurrence group(P<0.05).Receiver operating characteristic curves were plotted.The areas under the curves of preoperative serum UCH-L1 and GFAP levels for predicting postoperative glioma recurrence were 0.785 and 0.775,respectively.However,the efficacy of serum UCH-L1 and GFAP levels 3 d after surgery in predicting postoperative glioma recurrence was slightly lower compared with their preoperative levels.CONCLUSION UCH-L1 and GFAP efficiently reflected the development and recurrence of gliomas and could be used as potential indicators for the recurrence and prognosis of glioma.

Effects of P2Y_1 receptor on glial fibrillary acidic protein and glial cell line-derived neurotrophic factor production of astrocytes under ischemic condition and the related signaling pathways

Objective The present study aimed to explore the role of P2Y1 receptor in glial fibrillary acidic protein （GFAP） production and glial cell line-derived neurotrophic factor （GDNF） secretion of astrocytes under ischemic insult and the related signaling pathways. Methods Using transient right middle cerebral artery occlusion （tMCAO） and oxygen-glucose-serum deprivation for 2 h as the model of ischemic injury in vivo and in vitro, immunofluorescence, quantitative real-time reverse transcription-polymerase chain reaction （RT-PCR）, Western blotting, enzyme linked immunosorbent assay （ELISA） were used to investigate location of P2Y1 receptor and GDNF, the expression of GFAP and GDNF, and the changes of signaling molecules. Results Blockage of P2Y1 receptor with the selective antagonist N^6-methyl-2′-deoxyadenosine 3′,5′-bisphosphate diammonium （MRS2179） reduced GFAP production and increased GDNF production in the antagonist group as compared with simple ischemic group both in vivo and in vitro. Oxygen-glucose-serum deprivation and blockage of P2Y1 receptor caused elevation of phosphorylated Akt and cAMP response element binding protein （CREB）, and reduction of phosphorylated Janus kinase2 （JAK2） and signal transducer and activator of transcription3 （STAT3, Ser727）. After blockage of P2Y1 receptor and deprivation of oxygen-glucose-serum, AG490 （inhibitor of JAK2） reduced phosphorylation of STAT3 （Ser727） as well as expression of GFAP; LY294002, an inhibitor of phosphatidylinositol 3-kinase （PI3-K）, decreased phosphorylation of Akt and CREB; the inhibitor of mitogen-activated protein kinase kinase 1/2 （MEK 1/2） U0126, an important molecule of Ras/extracellular signal- regulated kinase （ERK） signaling pathway, decreased the phosphorylation of JAK2, STAT3 （Ser727）, Akt and CREB. Conclusion These results suggest that P2Y1 receptor plays a role in the production of GFAP and GDNF in astrocytes under transient ischemic condition and the related signaling pathways may be JAK2/STAT3 and PI3-K/Akt/CREB, respectively, and that crosstalk probably exists between them.

Expressions of nestin and glial fibrillary acidic protein in rat retina after optic nerve transection

AIM：To assess the expression of nestin and glial fibrillary acidic protein（GFAP）in rat retina after optic nerve transection.METHODS：Rats were randomly divided into normal control group,sham group and operation group,and used for establishing an animal model of optic nerve transection.Retinal specimen of each group was collected at 3,48h,7and 14d postoperative.Nestin and GFAP expressions on sagittal sections were analyzed by immunohistochemical staining,and protein extraction was analyzed by Western blot.RESULTS：Immunohistochemical analysis showed that nestin positive staining was rarely detected in normal control group and sham group,while sham group showed weak positive staining at 3h postoperative,the reaction gradually increased at 48h postoperative,and reached its maximum at 7d postoperative,and then decreased at 14d postoperative.Compared to the expression of GFAP,there was not statistically significant obvious difference among three groups（P〉0.05）.Result of Western blot method was consistent with that of immunohistochemical method.CONCLUSION：The expression of nestin increased in a time dependent fashion in Müller cells of retina following optic nerve transection,which was statistically significant,but there was no obvious difference in GFAP expression.The results indicate that an increase in colloid synthesis in retina following optic nerve transection can improve the retinal neurons’environment.

Effect of electroacupuncture on glial fibrillary acidic protein and nerve growth factor in the hippocampus of rats with hyperlipidemia and middle cerebral artery thrombus

Electroacupuncture(EA)has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia.However,there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects.In this study,EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed.Except for the normal group,rats in other groups were fed a high-fat diet throughout the whole experiment.Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks.Middle cerebral artery thrombus models were induced by pasting 50%FeCl3 filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group.EA1 group rats received EA at bilateral ST40(Fenglong)for 7 days before the thrombosis.Rats in the EA1 and EA2 groups received EA at GV20(Baihui)and bilateral ST40 for 14 days after model establishment.Neuronal health was assessed by hematoxylin-eosin staining in the brain.Hyperlipidemia was assessed by biochemical methods that measured total cholesterol,triglyceride,low-density lipoprotein and high-density lipoprotein in blood sera.Behavioral analysis was used to confirm the establishment of the model.Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region.The results demonstrated that,compared with the model group,blood lipid levels significantly decreased,glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups.The repair effect was superior in the EA1 group than in the EA2 group.These findings confirm that EA can reduce blood lipid,inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis.All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine,China(approval No.BUCM-3-2018022802-1002)on April 12,2018.

Hippocampal and cortical expression of gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein in pentylenetetrazol-induced chronic epileptic rats

BACKGROUND： Gamma-aminobutyric acid transporter plays an important role in gamma-aminobutyric acid metabolism, and is highly associated with epilepsy seizures. Pathologically, astrocytes release active substances that alter neuronal excitability, and it has been demonstrated that astrocytes play a role in epileptic seizures. OBJECTIVE： To observe changes in gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein expression in the hippocampus and cortex of the temporal lobe in rats with pentylenetetrazol-induced chronic epilepsy. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment was performed at the Department of Neurobiology, Third Military University of Chinese PLA between January 2006 and December 2007. MATERIALS： Pentylenetetrazol was purchased from Sigma, USA; rabbit anti-rat gammaaminobutyric acid transporter 1 and glial fibrillary acidic protein were from Chemicon, USA. METHODS： A total of 40 Sprague Dawley rats were divided into model and control groups. Rat models of chronic epilepsy were created by pentylenetetrazol kindling, and were subdivided into 3-, 7-, and 14-day kindling subgroups. MAIN OUTCOME MEASURES： Gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein expression, as well as the number of positive cells in the hippocampus and cortex of temporal lobe of rats, were determined by immunohistochemistry and Western blot analyses. RESULTS： Compared with the control group, the number of gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein -positive cells in the hippocampus and cortex of rats with pentylenetetrazol-induced epilepsy significantly increased, gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein expression increased after 3 days of kindling, reached a peak on day 7, and remained at elevated levels at day 14 （P〈 0.05）. CONCLUSION： Astrocytic activation and gamma-aminobutyric acid transporter 1 overexpression may contribute to pentylenetetrazol-induced epilepsy.

Scorpion ethanol extract and valproic acid effects on hippocampal glial fibrillary acidic protein expression in a rat model of chronic-kindling epilepsy induced by lithium chloride-pilocarpine

The present study analyzed the effects of ethanol extracts of scorpion on epilepsy prevention and hippocampal expression of glial fibrillary acidic protein in a lithium chloride-pilocarpine epileptic rat model. Results were subsequently compared with valproic acid. Results showed gradually- increased hippocampal glial fibrillary acidic protein expression following model establishment; glial fibrillary acidic protein mRNA expression was significantly increased at 3 days, reached a peak at 7 days, and then gradually decreased thereafter. Ethanol extracts of scorpion doses of 580 and 1 160 mg/kg, as well as 120 mg/kg valproic acid, led to a decreased number of glial fibrillary acidic protein-positive cells and glial fibrillary acidic protein mRNA expression, as well as decreased seizure grades and frequency of spontaneously recurrent seizures. The effects of 1 160 mg/kg ethanol extracts of scorpion were equal to those of 120 mg/kg valproic acid. These results suggested that the anti-epileptic effect of ethanol extracts of scorpion were associated with decreased hippocampal glial fibrillary acidic protein expression in a rat model of lithium chlofide-pilocarpine induced epilepsy.

Glial fibrillary acidic protein levels are associated with global histone H4 acetylation after spinal cord injury in rats

Emerging evidence has suggested global histone H4 acetylation status plays an important role in neural plasticity. For instance, the imbalance of this epigenetic marker has been hypothesized as a key factor for the development and progression of several neurological diseases. Likewise, astrocytic reactivity-a wellknown process that markedly influences the tissue remodeling after a central nervous system injury-is crucial for tissue remodeling after spinal cord injury（SCI）. However, the linkage between the above-mentioned mechanisms after SCI remains poorly understood. We sought to investigate the relation between both glial fibrillary acidic protein（GFAP） and S100 calcium-binding protein B（S100B）（astrocytic reactivity classical markers） and global histone H4 acetylation levels. Sixty-one male Wistar rats（aged ~3 months） were divided into the following groups： sham; 6 hours post-SCI; 24 hours post-SCI; 48 hours post-SCI; 72 hours post-SCI; and 7 days post-SCI. The results suggested that GFAP, but not S100B was associated with global histone H4 acetylation levels. Moreover, global histone H4 acetylation levels exhibited a complex pattern after SCI, encompassing at least three clearly defined phases（first phase： no changes in the 6, 24 and 48 hours post-SCI groups; second phase： increased levels in the 72 hours post-SCI group; and a third phase： return to levels similar to control in the 7 days post-SCI group）. Overall, these findings suggest global H4 acetylation levels exhibit distinct patterns of expression during the first week post-SCI, which may be associated with GFAP levels in the perilesional tissue. Current data encourage studies using H4 acetylation as a possible biomarker for tissue remodeling after spinal cord injury.

Dynamic changes of glial fibrillary acidic protein and nestin in the hippocampus of adult rat brain following ischemic vascular dementia

Vascular dementia produced by permanent ligation of bilateral common carotid arteries involves progressive deterioration of intellectual and cognitive function in rats, which are closely associated with the hippocampus. This study used immunohistochemical analysis to detect the expression of glial fibrillary acidic protein and nestin in the hippocampus in a vascular dementia model. The results revealed that both glial fibrillary acidic protein and nestin expression were increased 1 day after permanent ligation of the bilateral common carotid arteries, compared with a sham-operated group. The expression of glial fibrillary acidic protein peaked at 7 days post-surgery. The expression of nestin was a little weaker than that of glial fibrillary acidic protein, and peaked at 14 days （P 〈 0.01）. The expression of both proteins slightly decreased at 21 and 28 days, accompanied by recovery of cerebral blood flow. In conclusion, this study demonstrated that glial fibrillary acidic protein and nestin exhibited dynamic expression in the rat hippocampus after permanent ligation of bilateral common carotid arteries. This finding suggests that dynamic alterations in protein expression play an important role in the pathogenesis of vascular dementia.

Curcumin alters expression of glial fibrillary acidic protein and nestin following chronic cerebral ischemia

Astrocytes can alter their appearance and become reactive following chronic cerebral ischemia. In the present study, a rat model of chronic cerebral ischemia was treated with 50 and 100 mg/kg curcumin. Results showed that pathological changes of neuronal injury in hippocampal CA1 area of rats induced by chronic cerebral ischemia were attenuated, as well as upregulated expression of glial fibriliary acidic protein and nestin, in a dose-dependent manner.

Expression of vimentin and glial fibrillary acidic protein in central nervous system development of rats

Objective: To investigate the distribution and contents of vimentin(Vim) and glial fibrillary acidic protein(GFAP) immunoreactivities in the central nervous system(CNS)of normal newborn, adult and aged rats.Methods: In this study, thirty healthy and normal Sprague–Dawley rats were simply classified into three groups: Newborn(7 days aged), adult(5 months aged) and aged(24 months aged) rats. Brains and spinal cord were dissected and cut into frozen sections. The expression of Vim and GFAP in CNS were detected by confocal immunofluorescence.Results: In each group, Vim was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was highest in neuron-like cell of newborn rats, while Vim was mainly expressed in cell bodies in adult and aged rats. GFAP was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was in astrocytes of aged rats. In the third ventricle, Vim was detected in all groups, and only observed in neuron-like cells of newborn. Meanwhile, the GFAP expression showed no significant differences between adult and aged rats in this region. The co-localization of Vim and GFAP were mainly observed in hippocampus and cerebral cortex of newborn,but this co-localization was found in the third ventricle of the rats in all groups.Conclusion: Our data demonstrate for the first time that the expression of Vim and GFAP in the rat's CNS during development. This data may provide a foundation for the further mechanistic studies of these two main intermediate filaments during development of CNS.

Optic neuropathy and increased retinal glial fibrillary acidic protein due to microbead-induced ocular hypertension in the rabbit

AIM： To characterize whether a glaucoma model with chronic elevation of the intraocular pressure （IOP） was able to be induced by anterior chamber injection of microbeads in rabbits.METHODS： In order to screen the optimal dose of microbead injection, IOP was measured every 3d for 4wk using handheld applanation tonometer after a single intracameral injection of 10 μL, 25 μL, 50 μL or 100 μL microbeads （5×10^6 beads/mL; n=6/group） in New Zealand White rabbits. To prolong IOP elevation, two intracameral injections of 50 μL microbeads or phosphate buffer saline （PBS） were made respectively at days 0 and 21 （n=24/group）. The fellow eye was not treated. At 5wk after the second injection of microbeads or PBS, bright-field microscopy and transmission electron microscopy （TEM） were used to assess the changes in the retina. The expression of glial fibrillary acidic protein （GFAP） in the retina was evaluated by immunofluorescence, quantitative real-time polymerase chain reaction and Western blot at 5wk after the second injection of microbeads.RESULTS： Following a single intracameral injection of 10 μL, 25 μL, 50 μL or 100 μL microbead, IOP levels showed a gradual increase and a later decrease over a 4wk period after a single injection of microbead into the anterior chamber of rabbits. A peak IOP was observed at day 15 after injection. No significant difference in peak value of IOP was found between 10 μL and 25 μL groups （17.13±1.25 mm Hg vs 17.63±0.74 mm Hg; P=0.346）. The peak value of IOP from 50 μL group （23.25±1.16 mm Hg） was significantly higher than 10 μL and 25 μL groups （all P〈0.05）. Administration of 100 μL microbead solution （23.00±0.93 mm Hg） did not lead to a significant increase in IOP compared to the 50 μL group （P=0.64）. A prolonged elevated IOP duration up to 8wk was achieved by administering two injections of 50 μL microbeads （20.48±1.21 mm Hg vs 13.60±0.90 mm Hg in PBS-injected group; P〈0.05）. The bright-field and TEM were used to assess the changes of retinal ganglion cells （RGCs）. Compared with PBS-injected group, the extended IOP elevation was associated with the degeneration of optic nerve, the reduction of RGC axons （47.16%, P〈0.05） and the increased GFAP expression in the retina （4.74±1.10 vs 1.00±0.46, P〈0.05）.CONCLUSION： Two injections of microbeads into the ocular anterior chamber of rabbits lead to a prolonged IOP elevation which results in structural abnormality as well as loss in RGCs and their axons without observable ocular structural damage or inflammatory response. We have therefore established a novel and practical model of experimental glaucoma in rabbits.

Assessment of Neurotoxicity: Use of Glial Fibrillary Acidic Protein as a Biomarker

Diverse neurotoxic insults result in proliferation and hypertrophy of astrocytes. The hallmark of this response is enhanced expression of the major intermediate filament protein of astrocytes, glial fibrillary acidic protein (GFAP). These observations suggest that GFAP may be a useful biomarker of neurotoxicity. To investigate this possibility, we administered prototype neurotoxicants to experimental animals and assessed the effects of these agents on the tissue content of GFAP, as determined by radioimmunoassay. A review of the background, design, and results of these experiments are presented in this paper. Our findings indicate that GFAP is a sensitive and specific biomarker of neurotoxicity.

Endothelin-3 and glial fibrillary acidic protein expression in the frontal and parietal cortex of type-2 diabetic mice following ischemia-reperfusion injury

BACKGROUND： Patients with type-2 diabetes mellitus exhibit higher levels of plasma endothelin-1 （ET-1）. However, very few reports exist regarding altered endothelin-3 （ET-3） and ET-1 concentrations in brain tissue. OBJECTIVE： To observe expression changes of ET-3 and glial fibrillary acidic protein （GFAP） in the frontal and parietal cortex of type-2 diabetic mice following ischemia-reperfusion injury, with various reperfusion durations. DESIGN, TIME AND SETTING： Randomized controlled animal study. The experiment was conducted in the Xiangya Medical College of Central South University and the Third Xiangya Hospital between February 2002 and January 2003. MATERIALS： Sixty-six, adult, male, Kunming mice, weighing （30 ± 5） g, as well as rabbit anti-ET-3 polyclonal and rabbit anti-GFAP polyclonal antibodies, were provided by the Neurobiology Institute of Second Military Medical University in Japan. METHODS： Sixty-six mice were randomly divided into five groups： diabetes mellitus （DM, n = 6）, diabetes mellitus with ischemia-reperfusion （DM/IR, n = 24）, ischemia-reperfusion （IR, n = 24）, sham operation （SO, n = 6）, and control （n = 6）. MAIN OUTCOME MEASURES： Following ischemia-reperfusion for 1, 3, 5, and 10 days, respectively, expression of ET- 3 and GFAP was immunohistochemically measured in the frontal and parietal cortex. RESULTS： All 66 mice were included in the final result analysis. In the IR and DM/IR groups, ET-3- and GFAP-positive neurons increased in the frontal and parietal cortex in response to one day reperfusion, peaked at five days, and decreased at 10 days. ET-3 and GFAP expression was significantly greater in the DM/IR group after reperfusion for 1 day compared to the IR group. However, at other time points, there were no significant differences between the two groups. CONCLUSION： Brain ischemia-reperfusion injury results in overexpression of ET-3 and activation of astrocytes. Diabetes increases the number of ET-3- and GFAP-positive astrocytes in brain tissue of ischemia-reperfusion mice with the same reperfusion duration.

EXPRESSION OF NESTIN AND GLIAL FIBRILLARY ACIDIC PROTEIN IN DIFFERENT PERIOD AFT ER SPINAL INJURY IN ADULT RATS

Objective To study the ex pr ession of Nestin and glial fibrillary acidic protein (GFAP) in different period after spinal injury in adult rats. Methods Animal moels were cr eated by artery clamp. Expression of Nestin and GFAP in different period (1,3,5d ays;1-8 weeks) and different area(injury locus and its surrounding tissue ) afte r spinal injury were observed pathologicaly using immunofluorescence and LeicaQ5 00IW imaging processing system. Results There was expression of Nestin and GFAP in injured area 1 day after injury.The expression increased not only in in injured area but its sourrounding 3-7 days later and gradually got t o peak value. As the time went on, expression of Nestin and GFAP dereased. Conclusion Spinal injury can induce the expression of Nestin. Nerve stem cell has response to spinal injury. There is positive correlation between e xpression of Nestin and hyperplasia of reactivity astrocyte. Nerve stem cell may be invovled in the repair of central nervous system (CNS).

Autoantibodies to GFAP (glial fibrillary acidic protein) and to dopamine in patients with acute and chronic cerebrovascular disоrders

We have studied the level of autoantibodies to neurospecific proteins and neurotransmitters in patients with different forms of ischemic brain lesion. 49 patients with acute (ishemic stroke) and chronic cerebrovascular disease, 14 patients with ishemic heart disease and control group (35 healthy subjects) were investigated. The serum level of autoantibodies to glial fibrillary acidic protein (GFAP) and to dopamine (D) was determinated by ELISA. The content of autoantibodies to GFAP and D in patients with ischemic heart disease was practically identical. The patients with acute and chronic cerebrovas-cular diseases had the significally increased level of autoantibodies. The level of autoantibodies to GFAP in patients with acute vascular accidents (ischemic stroke) with favorable outcome was significantly higher than in patients with chronic cerebral ischemia. The obtained data allowed us to consider serum level of autoantibodies to GFAP as a marker of ischemic brain lesion, and to suppose further potential role of this autoantibodies in cerebrovascular disease progression.

Bilobalide inhibits the expression of aquaporin 1, 4 and glial fibrillary acidic protein in rat brain tissue after permanent focal cerebral ischemia

The present results demonstrated that in an adult rat model of permanent middle cerebral artery occlusion （pMCAO）, pretreatment with bilobalide reduced brain water content and infarct area, down-regulated aquaporin 1, 4 mRNA expression in brain edema tissue, then inhibited their synthesis in the striatum, in particular at the early stage of ischemia （at 8 hours after pMCAO）, inhibited glial fibrillary acidic protein expression, and lightened reactive gliosis. These data sug-gest that bilobalide attenuates brain edema formation due to reduced expression of aquaporins.

Inhibitory effect of synthetic small interfering RNAs on glial fibrillary acidic expression in astrocytes

BACKGROUND： Glial fibritlary acidic protein （GFAP） expression highly correlates with spinal glial scar formation, and is regarded as an important target for scar therapy. Efficient inhibition of expression could benefit recovery from spinal cord injury. OBJECTIVE： To investigate the inhibitory effects of synthetic small interfering RNAs （siRNAs） on astrocytic GFAP expression in rats. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment at the cellular and molecular level was performed at the First Hospital of Dalian Medical University between June 2005 and February 2006. MATERIALS： A total of 100 seven-day-old, Sprague Dawley rats were selected. GAPDH siRNA was purchased from Ambion, USA, And TransMessengerTM Transfection Reagent from DAKO, Carpinteria, CA. METHODS： Rat astrocytes were isolated and cultured. Three pairs of 21-nucleotide （nt） siRNAs specific to rats GFAP mRNA, 401,404 and 854, were synthesized and transfected in primary astrocytes at 1, 2, 3, and 4 g/L using TransMessengerTM Transfection Reagent. Non-transfected astrocytes served as the blank group. Cells transfected with siRNA were regarded as the negative control group, with GAPDH siRNA as the positive control group, and 401 siRNA, 404 siRNA, and 854 siRNA as experimental groups. MAIN OUTCOME MEASURES： GFAP mRNA and protein expression were assessed by RT-PCR and Western blot, respectively, at 24, 48, and 72 hours of culture. RESULTS： GFAP mRNA expression in the positive control group was significantly less than the negative control group （P 〈 0.01）. GFAP mRNA expression in astrocytes from three pairs of siRNA was significantly less than the blank group after 48 hours （P 〈 0.01 ）, while no differences were detected between the negative control and blank groups （P 〉 0.05）. GFAP protein expression was remarkably less in siRNA-transfected astrocytes compared to the blank control （P 〈 0.01 ）. CONCLUSION： Transfected siRNAs could significantly inhibit GFAP gene expression in astrocytes after 72 hours in culture.

视网膜静脉阻塞病人血清胶质纤维酸性蛋白和水通道蛋白4水平变化及其与预后的关系

目的检测视网膜静脉阻塞(RVO)病人血清中胶质纤维酸性蛋白(GFAP)、水通道蛋白4(AQP4)水平,探究二者表达水平与RVO病人预后的关系。方法回顾性选取2019年8月至2021年8月西安医学院第二附属医院收治的94例RVO病人为研究组,另取同期体检健康者85例为对照组。收集病人一般临床资料,对研究组和对照组的血清GFAP、AQP4水平进行检测;根据研究组病人预后情况将其分为预后良好组(39例)和预后不良组(55例);多因素logistic回归分析RVO病人预后的影响因素;绘制血清GFAP、AQP4对RVO病人预后评估的受试者操作特征曲线(ROC曲线)。结果研究组血清GFAP水平(3.56±0.74)μg/L显著高于对照组(1.85±0.41)μg/L,研究组血清AQP4水平(15.97±2.82)μg/L显著高于对照组(10.56±2.54)μg/L(P<0.05)。预后不良组血清GFAP水平(3.91±0.84)μg/L显著高于预后良好组(3.06±0.62)μg/L;预后不良组血清AQP4水平(17.25±3.29)μg/L显著高于预后良好组(14.18±2.18)μg/L(P<0.05)。预后良好组与预后不良组病人高血压史、空腹血糖、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)差异有统计学意义(P<0.05)。logistic回归分析显示,血清GFAP、AQP4、高血压史、空腹血糖、TC、TG、LDL-C和HDL-C均是RVO病人预后不良的影响因素(P<0.05)。血清中GFAP、AQP4、二者联合预测RVO病人预后的AUC分别是0.73、0.79、0.92,灵敏度分别为47.27%、78.85%、77.08%,特异度分别为89.74%、72.97%、97.14%,约登指数分别为0.370、0.518、0.742;二者联合优于GFAP、AQP4各自单独预测(均P<0.05)。结论RVO病人血清GFAP、AQP4水平显著升高,对病人的预后状况具有较高的预测效能,可为临床的合理干预和改善病人预后提供依据。

血清HIF-1α、NSE、GFAP及相关临床特征与新生儿缺氧缺血性脑病发生风险的关系分析

目的探讨血清低氧诱导因子-1α(HIF-1α)、神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)及相关临床特征与新生儿缺氧缺血性脑病(HIE)发生风险的关系。方法选取2020年1月—2023年1月苏州大学附属儿童医院收治的85例HIE患儿作为HIE组,另选取同期在该院出生的120例健康新生儿作为对照组,分析两组的临床资料并检测新生儿出生后3 d血清HIF-1α、NSE、GFAP水平。绘制受试者工作特征(ROC)曲线分析血清HIF-1α、NSE、GFAP水平预测新生儿HIE发病的价值;多因素逐步Logistic回归模型分析新生儿HIE发病的影响因素。结果与对照组相比,HIE组宫内窘迫、脐带异常、羊水污染、1 min Apgar评分≤7分的患儿比例较高(P<0.05),并且血清HIF-1α、NSE、GFAP水平较高(P<0.05);两组孕妇年龄、孕妇文化程度、胎龄、新生儿性别、出生体重、产次、剖宫产、胎膜早破比较,差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,HIF-1α、NSE、GFAP及三者联合预测新生儿HIE发病的敏感性分别为82.7%(95%CI:0.795,0.862)、78.7%(95%CI:0.705,0.849)、84.0%(95%CI:0.803,0.891)、85.3%(95%CI:0.788,0.922),特异性分别为85.3%(95%CI:0.816,0.907)、74.7%(95%CI:0.715,0.796)、72.0%(95%CI:0.692,0.771)、90.5%(95%CI:0.825,0.956),AUC分别为0.907(95%CI:0.884,0.930)、0.850(95%CI:0.816,0.884)、0.893(95%CI:0.827,0.959)、0.936(95%CI:0.905,0.967);多因素逐步Logistic回归分析显示,宫内窘迫[O^R=3.592(95%CI:2.017,6.397)]、脐带异常[O^R=4.905(95%CI:2.862,8.406)]、羊水污染[O^R=7.262(95%CI:3.603,14.637)]、1 min Apgar评分≤7分[O^R=3.139(95%CI:1.954,5.043)]、HIF-1α≥0.463 ng/mL[O^R=2.916(95%CI:1.422,5.980)]、NSE≥12.395μg/L[O^R=3.714(95%CI:1.955,7.056)]、GFAP≥3.962 ng/mL[O^R=3.556(95%CI:2.039,6.202)]均是新生儿HIE发病的危险因素(P<0.05)。结论宫内窘迫、脐带异常、羊水污染、出生后1 min Apgar评分低及血清HIF-1α、NSE、GFAP水平高是新生儿HIE发病的危险因素,临床通过检测血清HIF-1α、NSE、GFAP水平可为临床筛查HIE提供帮助,3项指标联合检测可进一步提高诊断价值。