单核细胞趋化蛋白1、颗粒蛋白前体、胶质细胞源性神经营养因子水平与阿尔茨海默病认知功能、日常生活能力相关性分析

目的 分析血清单核细胞趋化蛋白1(MCP-1)、颗粒蛋白前体(PGRN)、胶质细胞源性神经营养因子(GDNF)水平与阿尔茨海默病(AD)病人认知功能及日常生活能力的相关性,探讨其在AD中的诊断价值。方法 选取2019年7月至2021年6月入住苏州大学附属广济医院治疗的41例AD病人作为观察组,采用随机数字表法选取苏州大学附属广济医院同期经评估符合入组的41例健康志愿者作为对照组,运用酶联免疫吸附测定(ELISA)检测两组研究对象血清MCP-1、PGRN、GDNF水平;采用简易智力状态检查表(MMSE)及日常生活量表(ADL)评估其认知功能及日常生活能力。分析观察组病人血清MCP-1、PGRN、GDNF水平与MMSE、ADL评分的相关性;并通过受试者操作特征曲线(ROC曲线)分析MCP-1、PGRN、GDNF在AD中的诊断价值。结果 观察组MCP-1(321.7±51.1)ng/L、PGRN(42.6±8.5)μg/L水平较对照组MCP-1(242.6±35.2)ng/L、PGRN(26.5±6.1)μg/L显著增高,观察组GDNF(357.8±112.3)ng/L水平较对照组GDNF(...     

The GDNF-induced neurite outgrowth and neuronal survival in dissociated myenteric plexus cultures of the rat small intestine decreases postnatally

 Glial cell-line-derived neurotrophic factor (GDNF), a member of the transforming growth-factor- (TGF-) β-family, is an essential factor for the development of the enteric nervous system (ENS) during embryogenesis. In the present study, the effects of GDNF on postnatal ENS development were investigated using cultures of myenteric plexus from the small intestine of newborn albino rats of different developmental phases (P1, P7, P14). Myenteric plexus was dissociated and cultivated as mixed cultures of enteric neurons and glial cells. After seeding, the cultures were kept for 24 h or 7 days in serum-free medium containing various doses (1, 10, 100 ng/ml) of GDNF. The effect of the neurotrophic factor was evaluated using parameters such as cell size, neuronal survival, or neurite elongation. While neither glial-cell nor neuronal size was influenced by GDNF, there was an observable effect upon neuronal survival and neurite elongation. The cultures treated with GDNF displayed increased neurite outgrowth. The promoting effect was dose- and age-dependent, decreasing clearly during the early postnatal period. Already after 24 h, neuronal survival was increased in P1 and P7, but not in P14 cultures. In long-term cultures, a marked tendency to form cell aggregates and dense fiber networks was observed when treated with GDNF. These observations suggest that GDNF plays an important role not only in pre-, but also in postnatal development of the enteric nervous system. Received: 29 May 1998 / Accepted: 10 December 1998     

质粒型单纯疱疹病毒载体介导GDNF和GFP基因在体外培养的BHK细胞和大鼠脊髓、背根节神经元中的转移和表达

为了观察质粒型单纯疱疹病毒载体介导的外源性胶质细胞源性神经营养因子和绿色荧光蛋白基因在体外培养的金黄地鼠肾细胞以及脊髓神经元和背根节神经元中的转移和表达 ,本研究采用了以质粒型单纯疱疹病毒载体为基础构建的分别含有重组胶质细胞源性神经营养因子和绿色荧光蛋白基因的混合毒株 dv HSV-GDNF和 dv HSV-GFP感染体外培养的金黄地鼠肾细胞、脊髓神经元和背根节神经元。用免疫组织化学方法和荧光显微镜观测法分别检测了胶质细胞源性神经营养因子和绿色荧光蛋白基因的转移和表达。结果发现 :质粒型单纯疱疹病毒载体可以成功地将外源性胶质细胞源性神经营养因子和绿色荧光蛋白基因导入金黄地鼠的肾细胞以及脊髓神经元和背根节神经元中。提示质粒型单纯疱疹病毒载体可以作为转基因胶质细胞源性营养因子治疗脊髓损伤的转移载体 ,为脊髓损伤的基因治疗提供了实验基础。绿色荧光蛋白作为报告基因 ,也可因其适用广泛、观察简便等特性而得到更加广泛的应用。     

Familial form of Hirschsprung disease: Nucleotide sequence studies reveal point mutations in the RET proto‐oncogene in two of six families but not in other candidate genes

Hirschsprung disease (HSCR; McKusick 142623) or aganglionic megacolon is a frequent (1 in 5,000 live births) heritable disorder of the enteric nervous system. By haplotyping with a variety of microsatellite markers, by amplifying all 20 exons of the RET proto‐oncogene and by applying a direct DNA sequencing protocol, we have analyzed the DNA from HSCR patients in 6 different families. In one family with a joint occurrence of HSCR and FMTC (follicular medullary thyroid carcinoma), we have identified a mutation in codon 609 in one out of 6 cysteine residues encoded in exon 10 of the RET gene. This C609R point mutation has not previously been reported to cause HSCR. In 2 of the HSCR patients described here from different families, we have found a mutation in exon 2 (R77C) and a silent mutation in exon 3 (Y204Y), respectively, in the extracellular part of the RET proto‐oncogene. In introns 2 and 17 of the RET proto‐oncogene in 2 families, we have detected single nucleotide exchanges that are probably polymorphisms with unknown, if any, relations to HSCR. The DNA sequences of 5 further genes (GDNF, GDNFRα, EDN3, EDNRB, and NTN), that may contribute to the development of HSCR, have not shown mutations in the patients analyzed so far. In 2 of the reported families with several affected children and one grandchild, sequence analyses revealed no mutations in the coding regions of any of the candidate genes analyzed. Am. J. Med. Genet. 94:19–27, 2000. © 2000 Wiley‐Liss, Inc.     

Overexpression of the neuron-specific molecule bm88 in mouse neuroblastoma cells: Altered responsiveness to growth factors

Previous studies have shown that the BM88 antigen, a novel neuron-specific molecule, promotes the differentiation of mouse neuroblastoma (Neuro 2a) cells. In particular, stably transfected, with the BM88 cDNA, Neuro 2a cells overexpressing the BM88 antigen (Neuro2a-BM88 cells) are morphologically distinct from the nontransfected Neuro 2a cells; they exhibit enhanced process outgrowth and a slower rate of division. In this study we used Neuro2a and the morphologically differentiated Neuro 2a-BM88 cells to compare their responsiveness to growth factors. The growth factors we used were nerve growth factor (NGF), basic-fibroblast growth factor (b-FGF), and glial cell-line derived neurotrophic factor (GDNF). In addition, we used glial conditioned medium derived from either newborn mouse cerebral cortex (NBCC) or aged mouse cerebral hemispheres (MACH), as a source of normal glial factors. Because these cells express the cholinergic phenotype, we used choline acetyltransferase (ChAT) activity as a biochemical marker for comparison. A differential responsiveness to these factors was observed between Neuro 2a and Neuro 2a-BM88. The presence of NGF, 25 ng/ml, in the culture medium did not affect ChAT activity in either cell type. In contrast to NGF, in the presence of b-FGF, 5 ng/ml, the transfected cells, Neuro 2a-BM88, responded with a marked increase in ChAT activity. On the other hand, with GDNF, 1 ng/ml, only Neuro 2a cells showed an increase in ChAT activity. Finally, we found no response to the glial conditioned media, although these media contain several growth factors, including b-FGF. In conclusion, our findings show that overexpression of the neuron-specific antigen BM88 in neuroblastoma cells modifies their properties with respect to growth factor sensitivity, and, hence, the Neuro 2a and Neuro 2a-BM88 are suitable cell models to examine the role of growth factors in neuronal differentiation.     

不同时间针刺对局灶性脑缺血再灌注大鼠GDNF和bFGF蛋白表达的影响

目的观察局灶性脑缺血再灌注大鼠胶质细胞源性神经营养因子(GDNF)和碱性成纤维细胞生长因子(bFGF)蛋白表达的情况及不同时间针刺对二者的影响。方法采用线栓法复制大鼠大脑中动脉缺血再灌注模型,并分别在造模后6,12,24 h开始针刺,运用免疫组化法观察不同时间段治疗组与模型组缺血半暗带的GDNF和bFGF蛋白表达情况。结果在半暗带,bFGF和GDNF在缺血再灌注6 h都开始表达,随着时间延长,bFGF表达逐渐增加,而GDNF表达逐渐减弱;不同时间开始针刺的各组与相应时段的模型组相比,GDNF和bFGF蛋白表达均明显升高(P均<0.05);不同时间进行针刺的各组之间,12 h组、24 h组与6 h组比较二者均有显著性差异。结论针刺可通过促进神经营养因子的表达而对缺血半暗区的神经细胞具有保护作用,针刺介入的最佳时机应在脑缺血的早期。     

Past, present and future of A(2A) adenosine receptor antagonists in the therapy of Parkinson's disease

Several selective antagonists for adenosine A_2A receptors (A_2AR) are currently under evaluation in clinical trials (phases I to III) to treat Parkinson's disease, and they will probably soon reach the market. The usefulness of these antagonists has been deduced from studies demonstrating functional interactions between dopamine D₂ and adenosine A_2A receptors in the basal ganglia. At present it is believed that A_2AR antagonists can be used in combination with the dopamine precursor L-DOPA to minimize the motor symptoms of Parkinson's patients. However, a considerable body of data indicates that in addition to ameliorating motor symptoms, adenosine A_2AR antagonists may also prevent neurodegeneration. Despite these promising indications, one further issue must be considered in order to develop fully optimized antiparkinsonian drug therapy, namely the existence of (hetero)dimers/oligomers of G protein-coupled receptors, a topic that is currently the focus of intense debate within the scientific community. Dopamine D₂ receptors (D₂Rs) expressed in the striatum are known to form heteromers with A_2A adenosine receptors. Thus, the development of heteromer-specific A_2A receptor antagonists represents a promising strategy for the identification of more selective and safer drugs.     

GDNF促进帕金森病大鼠模型脑内移植的中脑源神经干细胞表达Pitx3、Nurr1和TH

目的:探讨胶质细胞源性神经营养因子(GDNF)作用下,中脑源神经干细胞(mNSCs)在帕金森病(PD)大鼠模型纹状体移植区表达垂体同源盒家族因子3(Pitx3)、孤儿核受体相关因子1(Nurr1)和酪氨酸羟化酶(TH)的变化.方法:mNSCs单独移植组、mNSCs+GDNF移植组和生理盐水假手术组的PD大鼠模型,移植4...     

在小鼠帕金森病模型中观察丙戊酸钠对多巴胺神经元及GDNF表达的影响

目的:通过建立小鼠帕金森病模型,观察丙戊酸钠(valproate,VPA)对中脑黑质多巴胺神经元及胶质源性神经营养因子(glial-derived neurotrophic factor,GDNF)表达的影响,以期探索丙戊酸钠的神经保护作用机制.方法:采用免疫组织化学方法检测小鼠中脑黑质多巴胺神经元;原位杂交方法检测纹...