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运用体外缺气损伤的细胞模型,研究GDNF对损伤的新生大鼠背根节神经元(DRG)是否有保护作用.采用原代培养的方法,用N1无血清培养基分离培养DRG神经元24 h后,加入含20 μg/mL GDNF的无血清培养液,然后液体石蜡封闭培养液,继续培养4,8,12,24 h,分别做MTT实验检测细胞活性OD值,台盼蓝染色记数,细胞总蛋白测定以及形态学观察突起生长状况.结果表明:①封闭12 h后,GDNF组的DRG细胞存活数同对照组相比有显著升高(P <0.01),其余各时间点均无显著性差异.缺气损伤8 h和12 h后,GDNF组的DRG细胞总蛋白较对照组显著升高 (** P <0.01,* P <0.05),其余各时间点均无显著性差异;②各时间点 DRG细胞活性及突起长度的变化差异均不显著.缺气损伤一定时间后,GDNF对新生大鼠DRG细胞的存活数及细胞总蛋白有明显的保护作用,而对细胞活性及突起的生长则没有明显的促进作用. 相似文献
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N1无血清培养基原代分离培养新生大鼠背根神经节（dorsal root ganglion DRG)神经元，用兴奋性毒素海仁酸（kainic acidKA)损伤体外培养细胞后，加入胶质源性神经营养因子（glial-derived neurotrophic factor,GDNF)作用。最后通过MTT法检测，细胞总蛋白测定，胎盘兰染色计数，形态学观察等方法研究分析GDNF对兴奋性毒素海仁酸损伤后大鼠DRG神经元的活性，存活及突起生长的影响，结果表明：GDNF对体外正常生长的DRG神经元的存活，活性及突起生长没有明显的促进作用。而在海仁酸损伤后，GDNF对其存活和活性有明显的促进作用。但对突起生长没有明显的影响。 相似文献
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以体外培养的仔猪睾丸支持细胞为研究对象，研究了FSH对GDNF蛋白的调节及其信号传导机制。结果显示：（1）外源性FSH以浓度和时间依赖性促进GDNF蛋白表达：FSH浓度为50ng／mL、作用1h时，GDNF水平达到最高；（2）用FSH（50ng／mL）或dbcAMP（100μmol／L）处理支持细胞，可迅速激活MEK激酶，随FSH刺激时间的延长，ERK1／2活性逐渐升高（0～2h）；（3）MEK1／2的抑制剂U0126可显著抑制FSH对GDNF的激活作用。结果表明，FSH在诱导GDNF蛋白表达的过程中，通过cAMP-PKA-ERK1／2级联通路，促进GDNF基因的表达。 相似文献
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采用简化的分离培养方法，将８日龄的鸡胚脑纹状体神经元在含白细胞介素－６（ＩＬ－６）１０μｇ／Ｌ的培养液内培养。经光镜和扫描电镜观察，纹状体神经元在培养液中能够迅速贴壁、存活、生长；随培养时间延长，试验组神经元的胞体明显大于对照组，神经元突起网亦变得更加稠密。 相似文献
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采用倒置相差显微镜、环境扫描电子显微镜对新生24h内SD大鼠大脑皮质原代培养的神经元进行了形态学观察，建立了大脑皮质神经元原代培养方法。应用2．5μg／mL阿糖胞苷（Ara-C）对培养3d的神经元进行24h干预，去除神经胶质细胞、成纤维细胞、神经干细胞等杂质细胞，以提高神经元产率；用Nissl’s染色法进行神经元鉴定；利用微量滴定（MTT）法测定了一个培养周期（约10d）的神经元活性分布。光学显微镜观察结果表明，培养至第4h绝大多数神经元已贴壁生长，胞体上可见1～2个细小突起；培养至第3d、第6d时，Nissl’s染色结果显示，加Aralc能显著提高神经元纯度（≥929／6）；MTT结合形态学观察结果表明，神经元培养至第6d时活性最强，细胞呈卵圆形、蜘蛛状、锥体状等多种形态，细胞胞体饱满，光晕明显，神经网络形成完整。 相似文献
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以原代培养的仔猪睾丸支持细胞（SC）为试验模型，研究了FSH对SC胶质细胞源性神经营养因子（GDNF）表达及其增殖的影响。结果显示，外源性添加FSH可促进GDNF蛋白表达，而且这种促进作用具有浓度和时间依赖关系，GDNF蛋白水平随着FSH浓度的增加而升高，FSH浓度为50ng／mL时，GDNF水平达到最高，然后逐渐降低；FSH（50ng／mL）作用30min，可显著促进GDNF蛋白的表达（P〈0．05），当FSH作用1h时，GDNF蛋白水平达到最高，随后逐渐降低。FSH可以时间-剂量依赖性促进SC增殖细胞核抗原（PCNA）的表达，FSH（50ng／mL）作用30min时，PCNA的表达显著增加（P〈0．05），作用1h达极显著水平（P〈0．01），至2h时，PCNA蛋白的表达水平达到最高，随后逐渐降低。提示，FSH可在体外条件下通过刺激SC表达GDNF进而促进其增殖。 相似文献
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为了探讨表没食子儿茶素没食子酸酚(tea polyphenol(-)-epigallocatechin gal-late,EGCG)对羟自由基损伤小鼠海马神经元的保护作用,采用原代细胞培养技术,分离培养18 d小鼠胚胎海马神经元,培养4 d后,加入FeSO4和H2O2进行处理,随后加入不同浓度的EGCG培养24 h,通过观察细胞的形态学变化,四甲基偶氮唑盐(methylthiazolydi-phenyl-tetrazolium bromide,MTT)比色法分析细胞活性,检测神经元中丙二醛(malondi-aldehyde,MDA)以及培养液中乳酸脱氢酶(lactate dehydrogenase,LDH)含量的变化,以探讨EGCG对羟自由基损伤神经元的保护作用。结果显示,EGCG能够抑制神经元MDA的产生,降低神经元细胞外液中LDH的含量,增加细胞的活性,从而进一步证明EGCG能够清除羟自由基产生,减轻羟自由基对神经元损伤,对神经元的存活有一定的保护作用,为其用于治疗中枢神经系统病理性损伤的打下实验基础。 相似文献
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为了研究家兔脑纹状体出血后一氧化氮合酶(NOS)活性和一氧化氮(NO)含量的变化及L-NNA和血塞通对其变化的影响,探讨NO在脑出血损伤中的作用机制及2种药物对脑神经元的保护作用.选取4月龄健康哈白兔56只,随机分为假手术对照组、模型组,脑纹状体出血血塞通治疗组及脑纹状体出血L-NNA治疗组,手术建立家兔脑出血模型,应用生化检测技术测定各组脑纹状体出血后不同时间出血侧纹状体NOS活性及NO含量.结果表明,模型组脑纹状体内NOS活性6h开始升高,3d达到高峰,随后逐渐下降,9d时基本恢复至正常水平；2治疗组纹状体NOS活性在各时间点极显著或显著低于模型组(P＜0.01或P＜0.05),并且L-NNA在6h～1d时降低脑纹状体NOS活性的程度好于血塞通治疗组(P＜0.05)；NO含量的变化与NOS活性的变化基本一致,二者成正相关；利用SABC免疫组织化学法方法检测各组脑纹状体神经型NOS(nNOS)阳性神经元,发现模型组纹状体nNOS阳性神经元密度增加,细胞着色加深,胞体截面积和最长突起长度变小,经过治疗后神经元密度降低,胞体截面积和最长突起长度显著改善(P＜0.05).结果提示:NO参与了脑出血损伤过程,高浓度的NO能发挥神经毒性作用损害脑组织；血塞通和L-NNA通过抑制NOS的活性,降低了脑纹状体NO的含量,对脑出血家兔的脑神经元具有明显的保护作用. 相似文献
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酵母培养物水溶物对丙二醛损伤的离体草鱼肠道黏膜细胞的保护作用 总被引：2，自引：0，他引：2
在培养液中加入不同浓度(4.94、9.89 μmol/L)的丙二醛(MDA)及酵母培养物水溶物,观察酵母培养物水溶物不同浓度(50、100、200 mg/L)、不同作用时间(3、6、9、12 h)下MDA损伤的离体草鱼肠道黏膜细胞生长、形态及相关酶活力的变化,以研究酵母培养物水溶物对MDA损伤的离体草鱼肠道黏膜细胞的保护作用.结果表明:培养液中添加4.94或9.89μmol/LMDA均显著抑制了离体草鱼肠道黏膜细胞生长(P＜0.05),致使贴壁细胞减少、细胞膜通透性增加,导致胞内酶漏出及细胞抗氧化酶活力降低.培养液中添加50、100、200 mg/L酵母培养物水溶物9h后可显著地促进细胞生长及提高细胞总蛋白含量(P＜0.05),改善细胞生长状态,其中以添加100 mg/L酵母培养物水溶物对4.94 μmol/L MDA损伤细胞的保护效果较好,其细胞生长状态能达到正常组水平.培养液中添加50、100、200酵母培养物水溶物能一定程度地降低培养液中MDA浓度,同时能降低MDA导致的细胞内酶漏出,提高MDA损伤的细胞抗氧化能力.由结果可知,对于4.94、9.89 μmol/L MDA对草鱼肠道黏膜细胞产生的损伤,培养液中添加50、100、200 mg/L酵母培养物水溶物12 h内对细胞均有保护作用,其中以添加100 mg/L酵母培养物水溶物对4.94 μmol/L MDA损伤细胞的保护作用最佳,使细胞生长状态接近正常组水平.酵母培养物水溶物可能通过增强细胞抗氧化能力途径起到对草鱼离体肠道黏膜细胞的保护作用. 相似文献
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Ovine scrapie was first recorded in Cyprus in 1985. Subsequently four dairy goats kept in two mixed flocks with affected sheep developed characteristic clinical signs similar to those seen in sheep. Fifteen goats from the two flocks were examined histologically and neurological lesions consistent with a diagnosis of scrapie were found in the four animals and in three others which had subsequently developed early neurological signs. These lesions were similar to those of naturally-affected sheep although neuronal degeneration and vacuolation were more severe in some cases. 相似文献
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Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1985,26(10):323-327
In late December 1984, 1853 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1984 and 35% of them responded. Veterinarians reported that 97,794 dogs were blood-tested to check for microfilariae and 1417 dogs (1.45% of those tested) were found with heartworm. Another 34 dogs were amicrofilaremic, but were diagnosed as having heartworm disease, to give the total number diagnosed in 1984 as 1451 (1.48%). Heartworm was reported from all provinces except Prince Edward Island and Newfoundland but most (1310) of the cases were in Ontario. In Quebec, 126 cases were reported mostly from west of Montreal.
Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 27% of the cases were observed with clinical signs of heartworm disease and 72% had a history of not having left Canada. Southwestern Ontario continued to be the primary focus of the infection.
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Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1984,25(9):347-350
In late December 1983, 2 800 veterinarians across Canada were sent a questionnaire in order to assess the status of heartworm disease in Canada in 1983 and 26% of them responded. Veterinarians reported that 59 504 dogs were blood-tested to check for microfilariae and 771 dogs (1.30% of those tested) were found with Dirofilaria immitis. Heartworm disease was diagnosed in all provinces except New Brunswick and Newfoundland but most (733) of the cases were in Ontario.
Heartworm disease was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 31% of the cases were observed with clinical signs of heartworm disease and 64% had a history of not having left Canada. Southwestern Ontario continues to be the focus of the infection and most of the dogs there had not left the province previously.
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Slocombe JO Villeneuve A 《The Canadian veterinary journal. La revue veterinaire canadienne》1993,34(10):630-633
In late November 1991, 1883 clinics in Canada were sent a questionnaire to assess the status of Dirofilaria immitis in dogs in 1991 and there was a 60.0% response. There were 344,031 dogs tested for heart-worm (HW), 627 were found infected and the prevalence of HW infection was 0.18%. There were 417 dogs with HW in Ontario, 116 in Manitoba, 38 in Quebec, 53 in British Columbia, three in Alberta, and one in Nova Scotia. In British Columbia, all of the infected dogs but one were from the Okanagan valley which, as from 1991, is a new focus of infection in Canada. Most dogs with HW had not been on preventive medication in 1990, and the prevalence among dogs tested and unprotected was 0.59%. That prevalence was considerably higher in endemic areas. Companion dogs, over three years of age and maintained primarily outdoors in rural areas, were most frequently infected. One cat was diagnosed with D. immitis and 33 dogs had Dipetalonema reconditium. 相似文献
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Owen J Slocombe D McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1986,27(9):324-328
In late December 1985, 1485 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1985 and 44% of them responded. Veterinarians reported that 137,300 dogs were blood-tested to check for microfilariae and 1210 dogs were found with heartworm. Another 36 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number diagnosed in 1985 as 1247 (0.91%).
Heartworm was reported from all provinces except Prince Edward Island, Newfoundland and Saskatchewan but most (1126) of the cases were in Ontario. Southwestern Ontario continued to be the primary focus of the infection in Canada. From Quebec, 91 cases were reported mostly from and around Montreal. From Manitoba, 19 cases were reported from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 28% of the cases were observed with clinical signs of heartworm disease and 78% had a history of not having left Canada.
相似文献19.
Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1989,30(6):504-508
In late November 1988, 1581 small and mixed animal clinics and institutional veterinarians across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1988, and 46% of them responded. Veterinarians reported that 181,577 dogs were blood-tested for heartworm disease and 367 dogs were found with D. immitis microfilariae. Another 60 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1988 as 441 (0.24%). 相似文献
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In late November 1989, 1732 clinics and institutional veterinarians were sent a questionnaire to assess the status of Dirofilaria immitis, and 51.7% responded. Of 247,716 dogs tested, 394 had D. immitis microfilariae and 51 were amicrofilaremic for a total of 445 cases and heartworm prevalence of 0.17%. Most (408) of these dogs had no preventive medication and the prevalence among dogs tested and unprotected was 1.01%. That prevalence was considerably higher in endemic areas. Thirty-seven dogs with heartworm had preventive medication. Heartworm was most frequent in companion dogs over three years of age maintained outdoors in rural areas. About 75% of the cases had never left Canada, 26% had clinical signs and 125 were not treated.
Heartworm was reported from British Columbia, Manitoba, Ontario, Quebec, Nova Scotia and Newfoundland, but 383 cases were in Ontario. South-western Ontario was the primary focus of infection. There were 33 cases in Quebec and 24 in Manitoba, mainly found in and around Metropolitan Montreal and Winnipeg respectively.
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