CAJ | 학술논문

目的观察乐果染毒对原代培养的大鼠皮层神经元的的毒性作用及损伤机制.方法体外原代培养SD大鼠皮层神经元6 d后,加入终浓度为1、5、10、50、100 μmol/L的乐果,于染毒后48 h收获细胞,取匀浆上清液用于氧化损伤及细胞内兴奋性氨基酸递质的测定,流式细胞术检测细胞凋亡.结果染毒48 h后,乐果浓度在5、10、50、100 μmol/L时,超氧化物歧化酶(SOD)活力[(1.04±0.02)、(0.99±0.02)、(0.96±0.02)、(0.91±0.02)U/mg pro]即明显降低,谷胱甘肽(GSH)含量[(219.35±6.79)、(205.6±6.29)、(194.06±2.63)、(93.68±7.56)mg/g pro]明显降低,丙二醛(MDA)含量[(21.22±0.29)、(24.01±0.34)、(27.15±1.02)、(32.91±1.39)nmol/mg pro]明显升高,与对照组比较,差异均有统计学意义(P<0.01);10、50、100 μmol/L剂量组天冬氨酸含量明显升高,1、5、10、50、100μmol/L剂量组谷氨酸含鼍明显升高,与对照组比较,差异均有统计学意义(P<0.01);1、10、100 μmol/L染毒组乐果对神经无细胞凋亡影响明显.乐果染毒后,神经元内MDA含量与兴奋性氨基酸(天冬氨酸、谷氨酸)含量呈正相关,相关系数分别为0.937和0.759;而与GSH含量呈负相关,相关系数分别为-0.868和-0.801,有统计学意义(P<0.01).结论乐果可引起神经元氧化损伤及兴奋性氨基酸递质含量变化,共同参与神经元的凋亡过程.
목적 관찰악과염독대원대배양적대서피층신경원적적독성작용급손상궤제.방법 체외원대배양SD대서피층신경원6 d후,가입종농도위1、5、10、50、100 μmol/L적악과,우염독후48 h수획세포,취균장상청액용우양화손상급세포내흥강성안기산체질적측정,류식세포술검측세포조망.결과 염독48 h후,악과농도재5、10、50、100 μmol/L시,초양화물기화매(SOD)활력[(1.04±0.02)、(0.99±0.02)、(0.96±0.02)、(0.91±0.02)U/mg pro]즉명현강저,곡광감태(GSH)함량[(219.35±6.79)、(205.6±6.29)、(194.06±2.63)、(93.68±7.56)mg/g pro]명현강저,병이철(MDA)함량[(21.22±0.29)、(24.01±0.34)、(27.15±1.02)、(32.91±1.39)nmol/mg pro]명현승고,여대조조비교,차이균유통계학의의(P<0.01);10、50、100 μmol/L제량조천동안산함량명현승고,1、5、10、50、100μmol/L제량조곡안산함타명현승고,여대조조비교,차이균유통계학의의(P<0.01);1、10、100 μmol/L염독조악과대신경무세포조망영향명현.악과염독후,신경원내MDA함량여흥강성안기산(천동안산、곡안산)함량정정상관,상관계수분별위0.937화0.759;이여GSH함량정부상관,상관계수분별위-0.868화-0.801,유통계학의의(P<0.01).결론 악과가인기신경원양화손상급흥강성안기산체질함량변화,공동삼여신경원적조망과정.
Objective To investigate the effect and mechanisms of dimethoate on the primary cul-tured cortical neuronal cell injury. Methods Cortical neuronal cells were isolated and cultured in serum free medium for 6 days in vitro,and 1,5,10,50 and 100 μmol/L dimethoate were added to the medium and intracellular SOD, MDA and GSH. The content of excitatory amino acid was measured after 48hours. Flow cytometry was used to detect the neuronal cell apoptusis. Results After 48 h ,the activity of SOD and the (205.6±6.29), (194.06±2.63),(93.68±7.56) mg/g pro],and the content of MDA increased obviously with 5,10,50 and 100 μmol/L dimethoate [(21.22±0.29),(24.01±0.34),(27.15±1.02),(32.91±1.39) nmol/mg pro];The content of Asp from 10 to 100 μmol/L dose group was higher than the control group and the con-tent of Glu from 1 to 100 μmol/L dose group was higher than the control group. The apoptosis rate had great significance between 1 to 100 μmol/L dose groups and control group. When treated with dirnethoate,MDA content in neuron was positively correlated with the content of EAAs with the increase of dimethoate. The correlative coefficient was 0.937 and 0.759 respectively(P<0.01),while it was negatively correlated with the content of GSH. The correlative coefficient was -0.868 and -0.801 respectively(P<0.01). Conclusion The oxidative damage and changes of excitatory amino acid content induced by Dimethoate contribute to the pri-mary cultured rat cortical neuron apoptosis.