AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate(Glu) in Hirschsprung's disease(HSCR).METHODS: Ninety children with HSCR and 50 children without HSCR matched fo...AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate(Glu) in Hirschsprung's disease(HSCR).METHODS: Ninety children with HSCR and 50 children without HSCR matched for similar nutritional status, age and basal metabolic index were studied. The expression and localization of neuroligin-1 and Glu were assessed using double-labeling immunofluorescence staining of longitudinal muscles with adherent myenteric plexus from the surgically excised colon of children with HSCR. Western blot analysis, quantitative real-time PCR(q RT-PCR) and immunohistochemistry were performed to evaluate the abundance of neuroligin-1 and Glu in different HSCR-affected segments(ganglionic, transitional, and aganglionic segments). Enzyme-linked immunosorbent assay(ELISA) was used to detect and compare serum Glu levels in the long-segment HSCR, short-segment HSCR and non-HSCR samples.RESULTS: Neuroligin-1 and Glu were co-expressed highest to lowest in the ganglionic, transi tional and aganglionic segments based on Western blot(neuroligin-1: 0.177 ± 0.008 vs 0.101 ± 0.006, 0.177 ± 0.008 vs 0.035 ± 0.005, and 0.101 ± 0.006 vs 0.035 ±0.005, P < 0.005; Glu: 0.198 ± 0.006 vs 0.115 ± 0.008, 0.198 ± 0.006 vs 0.040 ± 0.003, and 0.115 ± 0.008 vs 0.040 ± 0.003, P < 0.005) and q RT-PCR(neuroligin-1: 9.58 × 10-5 ± 9.94 × 10-6 vs 2.49 × 10-5 ± 1.38 × 10-6, 9.58 × 10-5 ± 9.94 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, and 2.49 × 10-5 ± 1.38 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, P < 0.005). Serum Glu level was the highest to lowest in the non-HSCR, short-type HSCR and long-type HSCR samples based on ELISA(in nmol/μL, 0.93 ± 0.31 vs 0.57 ± 0.25, 0.93 ± 0.31 vs 0.23 ± 0.16, and 0.57 ± 0.25 vs 0.23 ± 0.16, P < 0.005).CONCLUSION: Neuroligin-1 and Glu may represent new markers of ganglion cells, whose expression may correlate with the pathogenesis, diagnosis, differential diagnosis or classification of HSCR.展开更多
目的探讨亚慢性染铝对大鼠海马中神经连接蛋白1(NL1)与N-甲基-D-天冬氨酸受体(NMDAR)结合的影响,以及相关结合对大鼠长时程增强(LTP)的影响。方法取无特定病原体级健康雄性SD大鼠随机分为空白对照组、溶剂对照组和低、中、高剂量组,每...目的探讨亚慢性染铝对大鼠海马中神经连接蛋白1(NL1)与N-甲基-D-天冬氨酸受体(NMDAR)结合的影响,以及相关结合对大鼠长时程增强(LTP)的影响。方法取无特定病原体级健康雄性SD大鼠随机分为空白对照组、溶剂对照组和低、中、高剂量组,每组18只。空白对照组大鼠不予任何处理,溶剂对照组大鼠予剂量为1 m L/kg体质量的0.9%氯化钠溶液,低、中、高剂量组大鼠分别予质量浓度为0.41、0.81和1.62 mg/kg体质量的麦芽酚铝溶液,隔日腹腔注射,分别染毒1、2和3个月。染毒结束后,进行大鼠海马CA1区在体LTP测定;取大鼠海马,采用石墨炉原子吸收光谱法测定铝水平,采用免疫共沉淀和免疫印迹法测定与NL1结合的NMDAR1、NMDAR2B蛋白相对表达水平。结果溶剂对照组和低、中、高剂量组大鼠LTP均低于空白对照组(P<0.01);高剂量组大鼠LTP分别低于溶剂对照组和低、中剂量组(P<0.05)。低、中、高剂量组大鼠海马中铝水平均高于空白对照组和溶剂对照组(P<0.01)。各剂量组各时间点大鼠海马中与NL1结合的NMDAR1、NMDAR2B蛋白相对表达水平均低于同时间点空白对照组和溶剂对照组(P<0.01)。各剂量组2个月时间点的大鼠海马中与NL1结合的NMDAR1、NMDAR2B蛋白相对表达水平均低于同剂量组1个月时间点(P<0.01)。各剂量组3个月时间点大鼠海马中与NL1结合的NMDAR1和NMDAR2B蛋白相对表达水平均低于同剂量组1、2个月时间点(P<0.01)。结论麦芽酚铝可阻碍大鼠海马中NL1与NMDAR1和NMDAR2B的正常结合,进而影响NMDAR1和NMDAR2B对LTP的调节使其幅值下降,导致学习记忆损伤。展开更多
基金Supported by National Natural Science Foundation of China,No.81270720 and No.81471487
文摘AIM: To investigate the abundance and potential diagnostic significance of neuroligin-1 and glutamate(Glu) in Hirschsprung's disease(HSCR).METHODS: Ninety children with HSCR and 50 children without HSCR matched for similar nutritional status, age and basal metabolic index were studied. The expression and localization of neuroligin-1 and Glu were assessed using double-labeling immunofluorescence staining of longitudinal muscles with adherent myenteric plexus from the surgically excised colon of children with HSCR. Western blot analysis, quantitative real-time PCR(q RT-PCR) and immunohistochemistry were performed to evaluate the abundance of neuroligin-1 and Glu in different HSCR-affected segments(ganglionic, transitional, and aganglionic segments). Enzyme-linked immunosorbent assay(ELISA) was used to detect and compare serum Glu levels in the long-segment HSCR, short-segment HSCR and non-HSCR samples.RESULTS: Neuroligin-1 and Glu were co-expressed highest to lowest in the ganglionic, transi tional and aganglionic segments based on Western blot(neuroligin-1: 0.177 ± 0.008 vs 0.101 ± 0.006, 0.177 ± 0.008 vs 0.035 ± 0.005, and 0.101 ± 0.006 vs 0.035 ±0.005, P < 0.005; Glu: 0.198 ± 0.006 vs 0.115 ± 0.008, 0.198 ± 0.006 vs 0.040 ± 0.003, and 0.115 ± 0.008 vs 0.040 ± 0.003, P < 0.005) and q RT-PCR(neuroligin-1: 9.58 × 10-5 ± 9.94 × 10-6 vs 2.49 × 10-5 ± 1.38 × 10-6, 9.58 × 10-5 ± 9.94 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, and 2.49 × 10-5 ± 1.38 × 10-6 vs 7.17 × 10-6 ± 1.12 × 10-6, P < 0.005). Serum Glu level was the highest to lowest in the non-HSCR, short-type HSCR and long-type HSCR samples based on ELISA(in nmol/μL, 0.93 ± 0.31 vs 0.57 ± 0.25, 0.93 ± 0.31 vs 0.23 ± 0.16, and 0.57 ± 0.25 vs 0.23 ± 0.16, P < 0.005).CONCLUSION: Neuroligin-1 and Glu may represent new markers of ganglion cells, whose expression may correlate with the pathogenesis, diagnosis, differential diagnosis or classification of HSCR.
文摘目的探讨亚慢性染铝对大鼠海马中神经连接蛋白1(NL1)与N-甲基-D-天冬氨酸受体(NMDAR)结合的影响,以及相关结合对大鼠长时程增强(LTP)的影响。方法取无特定病原体级健康雄性SD大鼠随机分为空白对照组、溶剂对照组和低、中、高剂量组,每组18只。空白对照组大鼠不予任何处理,溶剂对照组大鼠予剂量为1 m L/kg体质量的0.9%氯化钠溶液,低、中、高剂量组大鼠分别予质量浓度为0.41、0.81和1.62 mg/kg体质量的麦芽酚铝溶液,隔日腹腔注射,分别染毒1、2和3个月。染毒结束后,进行大鼠海马CA1区在体LTP测定;取大鼠海马,采用石墨炉原子吸收光谱法测定铝水平,采用免疫共沉淀和免疫印迹法测定与NL1结合的NMDAR1、NMDAR2B蛋白相对表达水平。结果溶剂对照组和低、中、高剂量组大鼠LTP均低于空白对照组(P<0.01);高剂量组大鼠LTP分别低于溶剂对照组和低、中剂量组(P<0.05)。低、中、高剂量组大鼠海马中铝水平均高于空白对照组和溶剂对照组(P<0.01)。各剂量组各时间点大鼠海马中与NL1结合的NMDAR1、NMDAR2B蛋白相对表达水平均低于同时间点空白对照组和溶剂对照组(P<0.01)。各剂量组2个月时间点的大鼠海马中与NL1结合的NMDAR1、NMDAR2B蛋白相对表达水平均低于同剂量组1个月时间点(P<0.01)。各剂量组3个月时间点大鼠海马中与NL1结合的NMDAR1和NMDAR2B蛋白相对表达水平均低于同剂量组1、2个月时间点(P<0.01)。结论麦芽酚铝可阻碍大鼠海马中NL1与NMDAR1和NMDAR2B的正常结合,进而影响NMDAR1和NMDAR2B对LTP的调节使其幅值下降,导致学习记忆损伤。
