四波长内源光信号成像在GFAP-/-Vim-/-小鼠脑梗塞周围扩散性去极化中的应用

目的观测小鼠脑梗塞周围扩散性去极化（PIDs）的内源光信号变化特征,探索胶质纤维酸性蛋白（GFAP）和波形蛋白
（Vim）基因敲除对PIDs的影响。方法以GFAP和Vim基因敲除（GFAP-/-Vim-/-）小鼠及其野生型（GFAP+/+Vim+/+）小鼠为研究对
象,线栓法制备大脑中动脉梗塞（MCAO）模型,应用四波长内源光信号（OIS）成像技术监测两组动物4 h内PIDs的发作情况。
结果OIS成像显示PIDs在空间分布上表现为连续的、红蓝相间的弧形波,由发源处向四周缓慢播散;PIDs存在5种不同空间播
散类型：喙-尾播散类型、侧方-内侧播散类型、尾-喙播散类型、对侧播散类型和内侧-侧方播散类型。GFAP-/-Vim-/-小鼠和GFAP+/+
Vim+/+小鼠PIDs的发作次数、播散类型、速度及时程组间比较无统计学差异（P>0.05）。结论四波长OIS成像技术可获得PIDs
的高分辨率彩色影像,为更深入研究PIDs的时空特性创造了条件;GFAP和Vim基因敲除对MCAO模型制备后4 h内PIDs的发
作无显著影响。
     

成年大鼠脊髓损伤后反应性星形胶质细胞增生和巢蛋白表达的相关性及意义

目的 探讨脊髓损伤后反应性星形胶质细胞的增生和巢蛋白的表达情况,并观察其细胞类型及相关性.方法 利用动脉瘤夹压迫法建立成年大鼠脊髓损伤动物模型.利用BBB评分标准表、免疫组化、免疫荧光双标法与LeicaQ500IW图像处理系统分析和显示脊髓损伤后不同时期(1、3、5 d,1、2、4、6、8、w)、不同部位脊髓损伤的程度以及巢蛋白与胶质酸性纤维蛋白(GFAP)的表达变化及细胞类型.结果 BBB评分结果显示术后所有实验动物双后肢(HL)评分低至0~1min,随后逐渐上升,1～2周内恢复的幅度较大,以后恢复较缓慢,较正常对照组具统计学意义(P<0.05).甲苯胺蓝染色可见损伤区有核固缩、胞浆溶解、尼氏体模糊且染色较深的神经元.随着动物存活时间延长,损伤范围逐渐扩大,约1周后损伤范同不再扩大.正常对照组脊髓神经元胞体和突起轮廓清晰.免疫荧光双标染色及图像处理系统分析结果显示脊髓伤24h后可诱导损伤及邻近区域巢蛋白和GFAP高度表达,中央管周围室管膜区几乎均为巢蛋白/GFAP-细胞群.室管膜区以外的灰质和白质中几乎全是巢蛋白/GFAP 细胞群,3～7d逐渐达到高峰,之后逐渐减弱,在损伤后2周左右恢复至对照组水平(P<0.05).正常对照组在脊髓中央管室管膜区有少量巢蛋白/GFAP-细胞,在室管膜区以外的灰质和白质中偶可见染色强度较弱的巢蛋白/GFAP 共存细胞.结论 成年大鼠脊髓损伤可诱导巢蛋白和GFAP的表达.巢蛋白的表达和反应性星形胶质细胞增生呈正相关,且表达多相互共存;星形胶质细胞和神经干细胞对脊髓损伤都有反应,并可能参与中枢神经系统损伤修复.     

大豆异黄酮协同叶酸拮抗神经胶质细胞损伤的研究

目的探讨不同剂量大豆异黄酮单独干预、大豆异黄酮与叶酸联合干预对神经胶质细胞损伤的保护作用,并对其可能的作用机制进行探讨。方法选用24 h龄ICR小鼠,以肝微粒体酶混合物(S9)诱导环磷酰胺制备神经胶质细胞的损伤模型,采用单细胞凝胶电泳技术(彗星试验)和MTT细胞活性试验,观察大豆异黄酮、叶酸及两者联合作用后对神经胶质细胞损伤的拮抗作用。结果彗星试验中,与模型组比较,各干预组神经胶质细胞的彗星细胞拖尾率(P<0.05)和尾长值均明显降低(P<0.01);随着大豆异黄酮浓度的增加,大豆异黄酮单独和联合干预组的彗星细胞拖尾率和尾长值逐渐降低,但两者的差异无统计学意义(P=0.052)。MTT试验中,与模型组相比,各干预组的细胞活性明显增强(P<0.05),随着大豆异黄酮浓度的增加,大豆异黄酮单独或联合干预组的OD492值逐渐升高,尤其是大豆异黄酮和叶酸联合干预比相应的大豆异黄酮单独干预效果明显(P<0.05)。结论大豆异黄酮可以在一定程度上抑制神经胶质细胞损伤和增强细胞的活力,两者联合作用的效果更佳,且存在一定的剂量-反应关系。提示大豆异黄酮和叶酸对神经胶质细胞具有较好的协同作用。     

Autoimmune glial fibrillary acidic protein astrocytopathy mimicking acute disseminated encephalomyelitis: A case report

Introduction:Autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy is an increasingly recognized type of steroid-responsive autoimmune disease of the nervous system. Defined in 2016, it is associated with the presence of anti-GFAP immunoglobulinG in the serum or cerebrospinal fluid (CSF) of affected patients.Patient characteristics:Herein, we report a case of acute neurological symptoms, including headache, fever, confusion, and paralysis of the lower extremities. CSF analysis revealed lymphocytic pleocytosis and elevated protein levels, indicating acute disseminated encephalomyelitis, and the patient was given immunotherapy. Cranial magnetic resonance imaging showed multifocal T2/fluid-attenuated inversion recovery hyperintense signal changes in the periventricular white matter, and electromyography testing showed changes consistent with severe sensorimotor neuropathy, indicating the involvement of the brain and peripheral nerves.Diagnoses:Finally, a diagnosis of autoimmune GFAP astrocytopathy was confirmed due to the presence of GFAP-immunoglobulinG in the patient''s CSF.Interventions:The patient was treated with one course of intravenous immunoglobulin therapy, then followed with intravenous methylprednisolone (1.0 g/d for 3 days) and oral prednisolone.Outcomes:At 1 week after intravenous immunoglobulin therapy, his level of consciousness improved. However, flaccid paralysis persisted without substantial improvement.Conclusion:In conclusion, the provision of an accurate early diagnosis and appropriate treatment are crucial for improving the prognosis of patients with autoimmune GFAP astrocytopathy. Further, this case highlights the importance of recognizing the role of peripheral nerve involvement in GFAP autoimmunity.     

急性高眼压大鼠视网膜胶质细胞水通道蛋白-4的内化

目的:观察急性高眼压时大鼠视网膜胶质细胞水通道蛋白-4(AQP4)内化的时空变化及其规律,并探讨其与视网膜水肿的关系.方法:建模并分组;应用免疫荧光双标鉴定视网膜AQP4阳性细胞;观察AQP4与早期内涵体抗原1(EEA1)及晚期内涵体标记物甘露糖-6-磷酸受体(MPR)的共表达规律.结果:在正常视网膜内,表达AQP4的星形胶质细胞胞体位于节细胞层,Müller细胞的胞体位于内核层.正常及假手术组未见AQP4与EEA-1、MPR的共表达;高眼压作用不同时间,AQP4与EEA1共表达细胞的分布部位及其占AQP4阳性细胞比例都有改变,在高眼压作用60 min,AQP4与EEA1共表达细胞的比例达到最高值,AQP4与MPR共表达细胞分布的部位也有变化.结论:高眼压可诱导视网膜胶质细胞AQP4的内化,即AQP4经过内吞进入早期内涵体,再转运至晚期内涵体.AQP4的内化可能对肿胀的胶质细胞起保护作用.     

Optimal concentration and time window for proliferation and differentiation of neural stem cells from embryonic cerebral cortex: 5% oxygen preconditioning for 72 hours

Hypoxia promotes proliferation and differentiation of neural stem cells from embryonic day 12 rat brain tissue, but the concentration and time of hypoxic preconditioning are controversial. To address this, we cultured neural stem cells isolated from embryonic day 14 rat cerebral cortex in 5% and 10% oxygen in vitro. MTT assay, neurosphere number, and immunofluorescent staining found that 5% or 10% oxygen preconditioning for 72 hours improved neural stem cell viability and proliferation. With prolonged hypoxic duration(120 hours), the proportion of apoptotic cells increased. Thus, 5% oxygen preconditioning for 72 hours promotes neural stem cell proliferation and neuronal differentiation. Our findings indicate that the optimal concentration and duration of hypoxic preconditioning for promoting proliferation and differentiation of neural stem cells from the cerebral cortex are 5% oxygen for 72 hours.     

Lactulose enhances neuroplasticity to improve cognitive function in early hepatic encephalopathy

Lactulose is known to improve cognitive function in patients with early hepatic encephalopathy; however, the underlying mechanism remains poorly understood. In the present study, we investigated the behavioral and neurochemical effects of lactulose in a rat model of early hepatic encephalopathy induced by carbon tetrachloride. Immunohistochemistry showed that lactulose treatment promoted neurogenesis and increased the number of neurons and astrocytes in the hippocampus. Moreover, lactulose-treated rats showed shorter escape latencies than model rats in the Morris water maze, indicating that lactulose improved the cognitive impairments caused by hepatic encephalopathy. The present findings suggest that lactulose effectively improves cognitive function by enhancing neuroplasticity in a rat model of early hepatic encephalopathy.     

注射用丹参多酚酸治疗急性脑梗死的疗效及对认知功能、Hcy、NSE、S-100β、GFAP的影响

目的 探讨注射用丹参多酚酸治疗急性脑梗塞的效果及对认知功能、同型半胱氨酸（Hcy）、神经元特异性烯醇化酶（NSE）、S-100β、胶质纤维酸性蛋白（GFAP）的影响。方法 选取唐山市人民医院2017年10月-2018年10月收治的急性脑梗塞患者162例,采用随机数字表法分为两组（各81例）,对照组实施常规治疗,观察者在常规治疗基础上联用注射用丹参多酚酸100 mg/次,静脉滴注,1次/d,连续治疗14 d,于治疗前后检测Hcy、NSE、S-100β和GFAP,并评定患者认知功能、神经功能,比较两组临床疗效及不良反应。结果 两组患者治疗后Hcy、NSE、S-100β、GFAP、美国国立卫生研究院卒中量表（NHISS）评分均较治疗前降低（P<0.05）,简易精神状态评价量表（MMSE）评分、蒙特利尔认知评估量表（MOCA）评分较治疗前增加（P<0.05）;且观察组患者治疗后Hcy、NSE、S-100β、GFAP、NHISS评分低于对照组（P<0.05）,治疗后MMSE评分、MOCA评分高于对照组（P<0.05）。观察组患者总有效率高于对照组（P<0.05）;两组不良反应比较,差异无统计学意义（P>0.05）。结论 注射用丹参多酚酸治疗急性脑梗塞的效果显著,可改善认知功能和神经功能状况,并改善Hcy、NSE、S-100β、GFAP等指征。     

雪旺细胞外泌体影响胶质瘢痕形成修复小鼠脊髓损伤的实验研究

目的:探讨雪旺细胞外泌体（SCDEs）对小鼠脊髓损伤的修复作用。方法:从小鼠坐骨神经中提取原代雪旺细胞,并将雪旺细胞培养上清分次超速离心得到雪旺细胞外泌体。透射电镜鉴定外泌体 ,尾静脉注射外泌体,每周2次,每次3 μL/只,BMS评分观察小鼠行为学功能恢复,免疫荧光染色观察外泌体神经保护作用和胶质瘢痕变化情况。结果:SCDEs的直径范围为40~100 nm;与PBS组相比, SCDEs组行为学BMS评分增加（P<0.05）,神经学功能指标有所恢复（P<0.01）,胶质瘢痕增加（P<0.01）。结论:SCDEs可以促进损伤后胶质瘢痕的增加,修复脊髓损伤。     

Angiotensin II inhibits uptake of transferrin-bound iron but not non-transferrin-bound iron by cultured astrocytes

The existence of all components of the renin–angiotensin system (RAS) and the iron metabolism system, and the recent findings on the functions of angiotensin II (ANGII) in peripheral iron metabolism imply that ANGII might play a role in iron homeostasis by regulating expression of iron transport proteins in the brain. Here, we investigated effects of ANGII on uptake and release of iron as well as expression of cell iron transport proteins in cultured astrocytes. We demonstrated that ANGII could significantly inhibit transferrin-bound iron (Tf-Fe) uptake and iron release as well as the expression of transferrin receptor 1 (TfR1) and the iron exporter ferroportin 1 (Fpn1) in cultured astrocytes. This indicated that the inhibitory role of ANGII on Tf-Fe uptake and iron release is mediated by its negative effect on the expression of TfR1 and Fpn1. We also provided evidence that ANGII had no effect on divalent metal transporter 1 (DMT1) expression as well as non-transferrin-bound iron (NTBI) uptake in the cells. Our findings showed that ANGII has a role to affect expression of iron transport proteins in astrocytes in vitro and also suggested that ANGII might have a physiological function in brain iron homeostasis.