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   

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   

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   

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   

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   

1400W对脂多糖诱导少突胶质细胞前体死亡通路的阻断研究

目的：建立细菌脂多糖（LPS）诱导的2日龄感染型脑室周围白质软化（PVL）新生大鼠动物模型,探讨iNOS抑制剂1400W 对LPS诱导脑白质少突胶质细胞（OL）前体死亡的体内阻断效果。方法：2日龄新生大鼠随机分为假手术组、PVL组、1400W-即刻组、1400W-8h组、1400W-16h组以及1400W-24h组,分别在LPS注射后即刻、8 h、16 h以及24 h经皮下注射1400W 20 mg/kg。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、硝酸还原法检测一氧化氮（NO）含量、Western blot 法检测诱生型一氧化氮合酶（iNOS）表达量、免疫组织化学染色检测过氧亚硝酸盐（ONOO）含量以及OL前体数量。结果：LPS诱导可引起新生大鼠脑白质明显损伤,脑内iNOS表达明显上调,NO和ONOO-含量显著增加,脑白质内少突胶质细胞前体标记物(O4）标记的OL前体显著减少。与PVL组比较,1400W-即刻组、1400W-8h组以及1400W-16h组新生大鼠的脑白质病理均获明显改善,iNOS蛋白合成量、NO及ONOO-生成量均显著降低,OL前体数量明显增加（P<0.05）。1400W-24h组的各项检测结果与PVL组相似,均无明显改善。结论：体内研究确认1400W通过抑制iNOS的表达上调、减少脑内NO及ONOO-的生成量,从而阻断OL前体的死亡通路,发挥对脑白质的保护效果。在LPS诱导后16 h内应用1400W,均有良好的脑保护作用。［中国当代儿科杂志,2010,12（5）：357-362］     

胆道闭锁肝内iNOS及其调控因子异常表达与进行性肝损伤的相关性

目的 研究胆道闭锁(BA)患儿肝内诱导型一氧化氮合酶(iNOS)及其上下游调控因子表达情况,并探讨其与BA进行性肝损伤发生的关系.方法 应用免疫组织化学染色法对2002年10月至2007年3月在本院行Kasai手术的38例BA患儿与16例对照儿童肝组织iNOS表达情况进行研究;应用ELASA法对BA患儿和对照组外周血总一氧化氮(NO)代谢产物浓度进行测定;应用TUNEL法对BA患儿和对照组肝内胆管上皮凋亡指数进行测定;应用免疫印记法对BA患儿和对照组肝组织NF-κB表达进行半定量分析.结果 iNOS在BA患儿肝组织异常高表达,强度为0.30±0.08,而在对照组肝组织不表达.BA患儿外周血总NO代谢产物浓度为(90.40±12.46)mol/L,明显高于对照组的(63.67±5.78)μmol/L,且BA组总NO代谢产物浓度与血清ALT水平[(152.76±29.59)U/L]呈强正相关(r=0.97).BA组肝内胆管上皮凋亡指数(54.00±11.67)%远高于正常对照组(20.72±5.63)%,且与肝组织iNOS表达强度呈强正相关(r=0.99).NF-κB出在BA患儿肝组织中的表达(0.74±0.06)明显高于正常对照组(0.22±0.03),且与iNOS表达强度呈强正相关(r=0.97).结论 iNOS异常高表达在BA患儿进行性肝损伤中发挥重要作用,该作用是由iNOS及其上下游调控因子NF-κB、NO共同作用所实现.     

N-乙酰半胱氨酸对新生小鼠肝细胞内毒素损伤的保护作用

目的探讨N-乙酰半胱氨酸对新生小鼠肝细胞内毒素损伤时的保护作用,为新生小鼠内毒素肝损伤的防治研究提供理论依据。方法采用胶原酶消化组织块法分离新生小鼠的肝细胞,将细胞分内毒素处理组（LPS组）及N-乙酰半胱氨酸预处理组（NAC组）。LPS组培养基中加入LPS 10μg／ml;NAC组先在培养基中加入N-乙酰半胱氨酸5mmol／L,孵育1h后再加入LPS 10μg／ml。分别于加入LPS 0、6和12h收集各组肝细胞上清和肝细胞,每组12例,以生化法检测培养上清的丙氨酸氨基转移酶（alanine aminotransferase,ALT）和一氧化氮（nitric oxide,NO）的水平,并以逆转录聚合酶链反应（RT-PCR）法检测各组细胞诱导型一氧化氮合成酶（induced nitric oxide synthase,iNOS）mRNA的表达。结果LPS组在0、6、12h的ALT值分别为（21．1±4．78）U／L、（59．8±8．59）U／L、（89．6±15．30）U／L,NO水平分别为（1．6±0．31）μmol／L、（6．6±0．81）μmol／L、（7．8±1．01）μmol／L,iNOS mRNA水平分别为0．17±0．023、0．71±0．091、0．71±0．097。LPS组的ALT和NO、iNOS mRNA水平在6、12h较0h明显升高,差异有极显著统计学意义（P〈0．01）;NAC组上清中ALT和NO水平在0、6、12h分别为（20．6±5．98）U／L、（40．8±7．30）U／L、（55．4±5．48）U／L和（1．7±0．26）μmol／L、（3．2±0．71）μmol／L、（4．0±0．71）μmol／L,肝细胞iNOS mRNA的表达水平在0、6、12h分别为0．18±0．026、0．41±0．060、0．40±0．067。经NAC预处理后,在6、12h上清中ALT的水平与LPS组比较明显降低,而且NO水平以及肝细胞iNOS mRNA的表达与LPS组比较也明显降低,差异均有极显著统计学意义（P〈0．01）。结论NAC对新生小鼠肝细胞内毒素损伤有保护作用,NAC可能通过抑制LPS激活iNOS而发挥保护作用。     

高浓度氧对早产鼠肺一氧化氮合酶表达的影响

目的：明确高浓度氧对早产大鼠肺一氧化氮(nitric oxide, NO)合成及一氧化氮合酶(nitric oxide synthase, NOS)表达的影响,以探讨内源性NO在新生儿高氧肺损伤中的作用。方法：3日龄早产鼠随机分为空气组和高氧组,检测实验3 d及7 d时两组肺湿重/干重比值(W/D),肺组织病理学改变,支气管肺泡灌洗液中NO含量及诱导型NOS(iNOS),内皮细胞型NOS(eNOS)在肺内的分布和表达(免疫组织化学方法)。结果：3 d时高氧组表现为急性肺损伤：充血、出血、炎性渗出;7 d时,W/D值高于空气组(5.54±0.41) vs (5.00±0.15),(P<0.05),病理改变依然明显。与空气组相比,暴露3 d及7 d时,高氧组灌洗液中的NO含量(17.06±5.86)和(23.75±4.07) μmol/L较空气组(5.59±2.03)和(7.93±2.33) μmol/L明显上升(P均<0.01)。高氧组肺气道和肺泡上皮细胞、炎症细胞iNOS表达强阳性,强度高于空气组(P<0.01),且高氧7 d组高于3 d组(P<0.01)。与空气组比较,7 d时高氧组气道上皮细胞eNOS表达增加(P<0.05)。结论：高氧可上调早产大鼠肺炎症细胞、上皮细胞NOS的表达,促进NO合成,提示内源性NO介导参与了高氧诱导的肺损伤。     

Effect of 1400W on blocking lipopolysaccharide-induced microglial toxicity to preoligodendrocytes

Background  

The maternal-fetal infection/inflammation is believed to be the mechanism in the pathogenesis of periventricular leukomalacia (PVL). The activation of microglias (MGs) may contribute to preoligodendroglial damage. The present study was undertaken to explore the effect of N-[3-(aminomethyl) benzyl] acetamidine (1400W), a selective inhibitor of inducible nitric oxide synthase (iNOS), on the blockage of lipopolysaccharide (LPS)-induced microglial toxicity to preoligodendrocytes (preOLs).     

左旋精氨酸对模拟缺血再灌注乳兔培养窦房结细胞损伤的保护作用

目的 观察模拟缺血再灌注(IR)培养乳兔窦房结细胞(SANC)损伤及左旋精氨酸(L-Arg)的保护作用.方法 对SANC模拟低糖缺氧培养,添加L-Arg和L-Arg 加 L-NAME(L-Arg抑制剂)与培养细胞共同孵育,吖啶橙(AO)标记细胞凋亡,检测细胞内过氧化物歧化酶(SOD)活性、丙二醛(MDA)、总一氧化氮合酶(NOS)、氧化亚氮(NO)水平.细胞分为正常对照组、I120R120 组、I120R120加L-Arg组、I120R120加L-Arg加L-NAME组4组.结果 凋亡细胞体积明显缩小,核呈黄绿色,断裂为多个碎块状,由膜包裹着凸起于细胞表面呈黄绿色凋亡小体.与I120R120组凋亡率[(5.21±1.59)%]比较,I120R120加L-Arg组凋亡率[(8.70±3.10)%]显著降低(P<0.01);I120R120加L-Arg组细胞SOD活性[(46 820±14 450) U/g]较I120R120组 [(25 030±8 440) U/g]显著上升,而I120R120加L-Arg组MDA[(5.55±3.71)mmol/g]、NO[(3.65±1.02) U/g]和NOS[(3.73±0.24) μmol/L]较I120R120组MDA[(8.42±4.21)mmol/g]、NO[ (4.62±1.20) U/g]和NOS[(4.96±0.52) μmol/L]显著下降(Pa<0.05);而I120R120加L-Arg加L-NAME组各指标较I120R120组无显著性差异(Pa>0.05).结论 补充NO合成底物L-Arg能清除自由基,增加SOD活性,增强细胞抗氧化损伤能力,可能是阻断氧自由基所介导的细胞凋亡机制之一.     

自发性高血压大鼠主动脉中一氧化氮/一氧化氮合酶体系的变化

目的 探讨自发性高血压大鼠主动脉中一氧化氮(NO)/一氧化氮合酶(NOS)体系的变化.方法 随机选取健康雄性4周龄Wistar大鼠7只和4周龄自发性高血压大鼠7只,分别作为正常对照组及高血压组.4周龄及12周龄时检测二组大鼠血压.8周龄时取2组大鼠主动脉组织,采用硝酸还原酶法测定其NO水平,Western blot法检查其内皮型NOS(eNOS)以及诱导型NOS(iNOS)蛋白水平.用SPSS 13.0软件对数据进行统计学分析.结果 12周龄时高血压组大鼠血压明显高于正常对照组[(24.1±0 5) kPa vs (15.9±0.3) kPa](P<0.05).12周龄时高血压组大鼠主动脉组织中NO水平较正常对照组主动脉中NO水平低[(7.2±1 7) μmol·(g prot)-1 vs (17.1±5.3) μmol·(g prot)-1](P<0.05).12周龄时高血压组大鼠主动脉中eNOS蛋白表达量与正常对照组大鼠比较差异无统计学意义[(0.5±0.2) vs (0.5±0.3)](P>0.05);12周龄时高血压组大鼠主动脉中iNOS蛋白表达量显著低于正常对照组大鼠[(0.9±0.3) vs (1.5±0.5)](P<0.05).结论 自发性高血压大鼠主动脉中NO/NOS体系下调参与高血压的形成过程.     

不同剂量氨基胍对内毒素休克兔模型肾功能的影响

目的探讨早期应用不同剂量氨基胍对内毒素休克模型兔肾功能的保护作用及氨基胍治疗的剂量、时间依赖性.方法 40只新西兰白兔麻醉后被随机分成5组对照组、内毒素组、氨基胍一组、二组、三组.除对照组外,其他四组每只兔静脉推注大肠杆菌O55B5内毒素(LPS)400μg/kg,当平均动脉压下降为原来的30%时,休克诱导成功.氨基胍一、二、三组每只兔分别静脉注射30、50、100 mg/kg的氨基胍,对照组及内毒素组给予相应剂量的生理盐水.在注射内毒素前(T0)、休克时(T)、休克后第1(T1)、2(T2)、3(T3)、4(T4)、5(T5)、6(T6)小时记录尿量,在T、T2、T4、T6时间点测定血硝酸盐、亚硝酸盐(NO-3/NO-2,NO的代谢产物)、尿素氮(BUN)、肌酐(Scr)、尿视黄醇结合蛋白(RBP).结果 LPS可引起血NO-3/NO-2、BUN、Scr、尿RBP显著升高[分别从T点的(47±5)μmol/L、(5.8±1.5) mmol/L、(41±10)μmol/L、(240±61)ng/L,升高至T6 点的(160±18)μmol/L、(15.5±1.8) mmol/L、(166±23) μmol/L、(1580±180) ng/L, P均<0.01].尿量显著减少[从T0点的(17.6±2.8)ml降低到T6 点的(1.3±0.6)ml,P<0.01] .三个氨基胍组均可减轻 NO-3/NO-2、BUN、SCr、RBP升高的程度,增加尿量.氨基胍一、二、三组在T6点的NO-3/NO-2分别为(58±8)、(50±14)、(46±9)μmol/L,与内毒素组相比差异有极显著意义(P均<0.01);BUN分别为(8.2±2.9)、(7.5±1.9)、(5.5±1.8)mmol/L, 与内毒素组相比差异有极显著意义(P均<0.01);RBP分别为(350±60)、(272±72)、(248±103) ng/L,与内毒素组相比差异有显著意义( P分别<0.05、<0.05、<0.01);尿量分别为(11.1±2.4)、(12.1±1.3)、(17.1±2.4)ml,与内毒素组相比差异有极显著意义(P均<0.01).效果以大剂量氨基胍组为著.结论氨基胍可抑制NO的形成,且呈时间、剂量依赖性.氨基胍可不同程度地减轻LPS引起的尿量和肾功能改变.     

重症肌无力被动转移幼鼠的黏膜免疫耐受研究

目的 观察特异性耐受原-双类似物(Lys262-Ala207)对重症肌无力被动转移(PTMG)小鼠模型的黏膜免疫效果,探讨其作用机理及该方法在模型应用中的可行性.方法 将 C57BL/6幼年雌性小鼠60只分为3组:耐受组、模型组、对照组,各20只.耐受组和模型组建立PTMG模型.耐受组在致敏前10 d,每天经鼻腔滴人含耐受肽-双类似物25μg的PBS 50μl;模型组连续10 d经鼻腔滴入不含耐受肽的PBS 50μl;正常对照组不做特殊处理.观察实验各组小鼠的体重、临床评分、白细胞介素4(IL-4)、γ干扰素(IFN-γ)、TGF-a1及其他各项指标的改变情况.结果 模型组小鼠表现较为典型的PTMG症状;耐受组小鼠经耐受治疗有效,其临床症状较轻.血清乙酰胆碱受体抗体(AChRAb)含量耐受组为(16.01±1.09)mg/L,模型组为(28.12±1.28)mg/L,均高于对照组[(1.60±0.28)mg/L](t=44.37,70.27,P<0.01).耐受组外周血中TGF-B1[(437.19±1.93)ng/L]高于模型组[(175.63±3.12)ng/L](t=36.07,P<0.01),模型组IL-4、IFN-γ[(193.37±3.95)ng/L,(320.46±2.14)ng/L],均高于耐受组[(141.02±3.11)ng/L,(187.99±4.67)ng/L](t=37.20、51.69,P<0.01).各因子与对照组比较仍未达正常水平,差异有统计学意义(t=26.65、31.05、49.02,P<0.01).结论 双类似物鼻黏膜耐受对缓解PTMG有效,表现为:血清AChRAb含量降低,外周血中TGF-a1分泌增高,IL-4、IFN-γ分泌降低和临床症状的缓解等.同时该方法还具有使用方便、安全等特点.     

一氧化氮与肾小球疾病关系的初步研究

目的　了解一氧化氮（ＮＯ）与肾小球疾病发病关系。方法　采用Ｇｒｉｅｓｓ 硝酸盐还原法对４４ 例急性肾炎,３２ 例肾病综合征,１８ 例紫癜性肾炎进行了血清亚硝酸／硝酸盐（ＮＯ２－／ＮＯ３ －） 测定,并以２８ 例健康儿童作对照。结果　疾病组血清ＮＯ２ －／ＮＯ３ － 的浓度（ 急性肾炎：７０ ．８ ±３４．７ μｍｏｌ／Ｌ;肾病综合征：６６ ．６ ±２７．９;紫癜性肾炎：４７ ．４ ±２１．４ μｍｏｌ／Ｌ） 显著高于对照组（３０．３ ±８ μｍｏｌ／Ｌ,Ｐ＜０ ．０１）,疾病急性期高于缓解期（ Ｐ＜ ０．０５）,伴有感染者（９３ ．５ ±３２．９ μｍｏｌ／Ｌ） 高于无感染者（４８ ．７±１４ μｍｏｌ／Ｌ,Ｐ＜ ０．０１）。肾病综合征血清ＮＯ２－／ＮＯ３ － 浓度与胆固醇水平呈负相关（ Ｐ＜ ０ ．０１） ,与血浆白蛋白及尿蛋白定量无相关（Ｐ＞ ０．０５） 。结论　ＮＯ 可能参与这３ 种肾小球疾病发病及病理损伤过程。