iNOS抑制剂阻断由细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体毒性作用的研究

objective To explore the toxicity of LPS-induced activated microglia to preoligodendrocytes(preOLs)and the effect of 1400W,a selective inhibitor of inducible nitric oxide synthetase(iNOS),on the blockage of the toxicity.Methods Co-cultured micmglia and preOLs obtained from two-day-old Sprague-Dawley(SD) rats were divided into three groups:co-culture control group,co-culture LPS group and co-cultttre LPS plus 1400W group.After cultured ceils were induced by LPS (100 ng/ml)for 48 hours,the concentration of nitric oxide(NO) was measured by nitric acid-deoxidize-colorimetry,the level of peroxynitrite(ONOO-)was determined by immunocytochemistry,and the synthetic level of iNOS was deleted by Western blotting,respectively.The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously.Data were analyzed with SPSS 11.0 software.Results Compared to co-culture control group,there was significant increase in levels of NO[(82.27±3.41)μmol/L vs(167.86±9.87)μmol/L,t=8.593,P<0.01],ONOO-[(6.14±1-1.27)x 107/L vs.(34.38±7.75)×107/L,t=5.892,P<0.01],and iNOS[(0.18±0.027) vs.(0.79±0.068),t=9.26,P<0.01] induced by LPS in co-culture LPS group,and with a higher apoptotic rate of preOLs[(6.73±1.39)% vs.(24.77±2.05)%,t:12.619,P<0.01].However.all levels of NO[(69.55±5.07)μmol/L,t=8.896,P相似文献   

iNOS抑制剂阻断由细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体毒性作用的研究

objective To explore the toxicity of LPS-induced activated microglia to preoligodendrocytes(preOLs)and the effect of 1400W,a selective inhibitor of inducible nitric oxide synthetase(iNOS),on the blockage of the toxicity.Methods Co-cultured micmglia and preOLs obtained from two-day-old Sprague-Dawley(SD) rats were divided into three groups:co-culture control group,co-culture LPS group and co-cultttre LPS plus 1400W group.After cultured ceils were induced by LPS (100 ng/ml)for 48 hours,the concentration of nitric oxide(NO) was measured by nitric acid-deoxidize-colorimetry,the level of peroxynitrite(ONOO-)was determined by immunocytochemistry,and the synthetic level of iNOS was deleted by Western blotting,respectively.The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously.Data were analyzed with SPSS 11.0 software.Results Compared to co-culture control group,there was significant increase in levels of NO[(82.27±3.41)μmol/L vs(167.86±9.87)μmol/L,t=8.593,P<0.01],ONOO-[(6.14±1-1.27)x 107/L vs.(34.38±7.75)×107/L,t=5.892,P<0.01],and iNOS[(0.18±0.027) vs.(0.79±0.068),t=9.26,P<0.01] induced by LPS in co-culture LPS group,and with a higher apoptotic rate of preOLs[(6.73±1.39)% vs.(24.77±2.05)%,t:12.619,P<0.01].However.all levels of NO[(69.55±5.07)μmol/L,t=8.896,P相似文献   

未成熟脑白质缺血性损伤和谷氨酸异常信号传输

目的探讨缺血诱导谷氨酸异常信号传输对未成熟脑白质的损伤作用。方法分别制备2日龄新生大鼠少突胶质细胞(OL)前体氧糖剥夺(OGD)细胞模型和缺血型脑室周围白质软化(PVL)动物模型。于OGD后24小时,收集细胞及上清液,应用高效液相色谱仪测定OL前体胞外谷氨酸浓度,流式细胞仪检测胞内钙离子浓度以及OL前体凋亡率。新生大鼠于造模后第5天处死取脑,分别进行光镜下脑白质病理评估、免疫组化法检测髓鞘碱性蛋白(MBP)阳性成熟OL百分比以及电镜下脑白质髓鞘化评估。结果体外与无氧糖剥夺的正常组OL前体相比,OGD组OL前体胞外谷氨酸呈大量蓄积(P0.01),胞内钙离子浓度显著增加(P0.01),OL前体的凋亡率和坏死率亦明显增高(P0.01,P0.05)。体内与假手术(Sham)组正常新生大鼠相比,PVL组所有大鼠脑白质均显现轻度或重度的病理改变,脑白质内MBP阳性成熟OL明显减少,髓鞘亦形成不良,表现为髓鞘数目明显减少和髓鞘厚度明显变薄(P均0. 01)。结论未成熟脑白质具有与成熟脑白质相似的谷氨酸信号传输特点。缺血诱导未成熟脑白质内谷氨酸异常信号传输。     

iNOS抑制剂阻断由细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体毒性作用的研究

objective To explore the toxicity of LPS-induced activated microglia to preoligodendrocytes(preOLs)and the effect of 1400W,a selective inhibitor of inducible nitric oxide synthetase(iNOS),on the blockage of the toxicity.Methods Co-cultured micmglia and preOLs obtained from two-day-old Sprague-Dawley(SD) rats were divided into three groups:co-culture control group,co-culture LPS group and co-cultttre LPS plus 1400W group.After cultured ceils were induced by LPS (100 ng/ml)for 48 hours,the concentration of nitric oxide(NO) was measured by nitric acid-deoxidize-colorimetry,the level of peroxynitrite(ONOO-)was determined by immunocytochemistry,and the synthetic level of iNOS was deleted by Western blotting,respectively.The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously.Data were analyzed with SPSS 11.0 software.Results Compared to co-culture control group,there was significant increase in levels of NO[(82.27±3.41)μmol/L vs(167.86±9.87)μmol/L,t=8.593,P<0.01],ONOO-[(6.14±1-1.27)x 107/L vs.(34.38±7.75)×107/L,t=5.892,P<0.01],and iNOS[(0.18±0.027) vs.(0.79±0.068),t=9.26,P<0.01] induced by LPS in co-culture LPS group,and with a higher apoptotic rate of preOLs[(6.73±1.39)% vs.(24.77±2.05)%,t:12.619,P<0.01].However.all levels of NO[(69.55±5.07)μmol/L,t=8.896,P相似文献   

细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体的毒性作用

已有实验证实,脑白质病变如多发性硬化症和脑室周围白质软化等一些神经病变的发病机制,均涉及到局部小胶质细胞的激活。 目的：探讨细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体的毒性作用。 方法：取2 d龄SD大鼠脑内小胶质细胞和少突胶质细胞前体共培养,分为共培养对照组和共培养脂多糖组。对共培养细胞经脂多糖100 μg/L 诱导48 h,分别应用硝酸还原比色法检测一氧化氮含量,还原-比色法检测超氧阴离子含量,免疫细胞染色法检测过氧亚硝酸盐含量,Hochest33342/PI荧光染色法观察细胞死亡的形态学改变,以及流式细胞仪检测细胞成活率。 结果与结论：与共培养对照组比较,脂多糖诱导导致共培养细胞内一氧化氮、超氧阴离子、过氧亚硝酸盐含量均明显增加,少突胶质细胞前体的凋亡率亦显著增加。通过体外观察,确认脂多糖诱导少突胶质细胞前体的死亡通路,是由于脂多糖诱导小胶质细胞激活,导致小胶质细胞表达一氧化氮合酶和活化NADPH氧化酶,致使其产物一氧化氮和超氧阴离子大量生成,两者进一步反应生成大量的毒性因子过氧亚硝酸盐,作用于少突胶质细胞前体,从而导致少突胶质细胞前体的死亡。     

小胶质细胞和少突胶质细胞前体的培养和鉴定

目的 探讨新生大鼠脑组织小胶质细胞(MG)和少突胶质细胞(OL)前体的分离和体外培养方法 . 方法 取新生2 d SD大鼠脑组织,体外原代培养混合胶质细胞7 d后,分别采用"改良振荡伴差速贴壁"法和"营养缺失伴振荡"法纯化培养MG和OL前体,并分别应用免疫荧光染色异凝集素-B4(IB4)和OL前体标记物(O4)进行鉴定.结果 混合胶质细胞培养7 d后呈明显三层增长,其中MG位于上层,星型胶质细胞位于底层,两者之间为2型少突星型(O2A)祖细胞.纯化培养后OL前体胞体呈小圆形,有双极或三极突起,MG则以阿米巴形、圆形居多,或边缘呈毛刺状.免疫荧光染色IB4显示绿色荧光,MG纯度达到90%以上.免疫荧光染色O4显示棕黄色荧光,OL前体纯度达到95%以上. 结论 采用"改良振荡伴差速贴壁"法以及"营养缺失伴振荡"法分别成功获取大量纯度高、活力好的MG和OL前体.     

iNOS抑制剂阻断由细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体毒性作用的研究

objective To explore the toxicity of LPS-induced activated microglia to preoligodendrocytes(preOLs)and the effect of 1400W,a selective inhibitor of inducible nitric oxide synthetase(iNOS),on the blockage of the toxicity.Methods Co-cultured micmglia and preOLs obtained from two-day-old Sprague-Dawley(SD) rats were divided into three groups:co-culture control group,co-culture LPS group and co-cultttre LPS plus 1400W group.After cultured ceils were induced by LPS (100 ng/ml)for 48 hours,the concentration of nitric oxide(NO) was measured by nitric acid-deoxidize-colorimetry,the level of peroxynitrite(ONOO-)was determined by immunocytochemistry,and the synthetic level of iNOS was deleted by Western blotting,respectively.The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously.Data were analyzed with SPSS 11.0 software.Results Compared to co-culture control group,there was significant increase in levels of NO[(82.27±3.41)μmol/L vs(167.86±9.87)μmol/L,t=8.593,P<0.01],ONOO-[(6.14±1-1.27)x 107/L vs.(34.38±7.75)×107/L,t=5.892,P<0.01],and iNOS[(0.18±0.027) vs.(0.79±0.068),t=9.26,P<0.01] induced by LPS in co-culture LPS group,and with a higher apoptotic rate of preOLs[(6.73±1.39)% vs.(24.77±2.05)%,t:12.619,P<0.01].However.all levels of NO[(69.55±5.07)μmol/L,t=8.896,P相似文献   

iNOS抑制剂阻断由细菌脂多糖诱导活性小胶质细胞对少突胶质细胞前体毒性作用的研究

objective To explore the toxicity of LPS-induced activated microglia to preoligodendrocytes(preOLs)and the effect of 1400W,a selective inhibitor of inducible nitric oxide synthetase(iNOS),on the blockage of the toxicity.Methods Co-cultured micmglia and preOLs obtained from two-day-old Sprague-Dawley(SD) rats were divided into three groups:co-culture control group,co-culture LPS group and co-cultttre LPS plus 1400W group.After cultured ceils were induced by LPS (100 ng/ml)for 48 hours,the concentration of nitric oxide(NO) was measured by nitric acid-deoxidize-colorimetry,the level of peroxynitrite(ONOO-)was determined by immunocytochemistry,and the synthetic level of iNOS was deleted by Western blotting,respectively.The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously.Data were analyzed with SPSS 11.0 software.Results Compared to co-culture control group,there was significant increase in levels of NO[(82.27±3.41)μmol/L vs(167.86±9.87)μmol/L,t=8.593,P<0.01],ONOO-[(6.14±1-1.27)x 107/L vs.(34.38±7.75)×107/L,t=5.892,P<0.01],and iNOS[(0.18±0.027) vs.(0.79±0.068),t=9.26,P<0.01] induced by LPS in co-culture LPS group,and with a higher apoptotic rate of preOLs[(6.73±1.39)% vs.(24.77±2.05)%,t:12.619,P<0.01].However.all levels of NO[(69.55±5.07)μmol/L,t=8.896,P相似文献   

家庭化管理模式在慢性退缩精神病患者中的应用

慢性退缩精神病患者在本院住院患者中占很大一部分比例,这些患者由于长期住院且病程长、病情反复而导致精神日趋衰退,传统的药物疗效不佳。患者生活自理能力差或无、社会功能明显减退、体质差、年龄普遍老化且多伴有慢性躯体疾病,为提高他们的生活质量,最大限度减轻精神残疾,充分挖掘患者内在的潜能,减少由此带来的一系列问题,本院于2002年5月为30例慢性退缩精神病患者建立康复病房,对他们实施引导和启蒙相结合的家庭化管理,取得了较好的效果,报告如下．     

细菌溶解产物预防支气管哮喘儿童并发呼吸道感染的临床疗效观察

目的观察细菌溶解产物对于支气管哮喘儿童并发急性呼吸道感染的临床疗效和安全性。方法采用前瞻性研究,选取2016年1月至2018年12月上海交通大学医学院附属新华医院收治的120例支气管哮喘缓解期患儿,采用随机方法将患儿平均分为两组:对照组和治疗组,每组各60例。对照组患儿仅采用常规吸入性糖皮质激素治疗,治疗组患儿在常规吸入性糖皮质激素治疗的基础上,给予细菌溶解产物治疗(3.5 mg/d,每月10 d,连续3个月),连续随访1年,比较两组患儿治疗9个月、12个月时急性呼吸道感染发生次数、哮喘累计发作次数、喘息累计发作天数及鼻塞发作次数,记录两组患儿血液淋巴细胞亚群及用药后不良反应发生情况。结果治疗组患儿的急性呼吸道感染发生次数(2.89±1.07、4.30±1.06)低于对照组(3.50±1.282、5.00±1.262),差异具有统计学意义(P<0.01);治疗组患儿哮喘发作次数(0.36±0.65、0.44±0.71)低于对照组(0.68±1.12、1.00±1.62),差异具有统计学意义(P<0.01);治疗组鼻塞发作次数(0.36±0.65、0.45±0.78)低于对照组(0.36±0.65、0.76±0.98),差异具有统计学意义(P<0.01)。治疗组患儿3个月随访患儿血清CD3^+%、CD4^+%、CD4^+/CD8^+水平高于对照组CD3^+%、CD4^+%、CD4^+/CD8^+,差异具有统计学意义(P<0.01),但两组患儿的CD8^+%水平比较,差异无统计学意义(P>0.05)。细菌溶解产物治疗期间共报道5例不良事件,均轻微短暂,未导致受试药品停用。结论应用细菌溶解产物可以预防支气管哮喘儿童并发呼吸道感染,可显著减少患儿呼吸道感染发生次数、喘息发作次数、鼻塞发作次数和喘息累计发作天数,增强细胞免疫机能,且安全性较高。