Effects Of N-methyl-D-aspartate Receptor And Ca～(2+) In Acute Intoxicated Encephalopathy Induced By 1,2-dichloroethane

Objective To study role of N-methyl-D-aspartate receptor and Ca²⁺in acute intoxicated encephalopathy induced by 1,2-dichloroethane(1,2-DCE)in vitro.Methods Neurocytes of new born rats were cultured in vitro. which were administrated with different dose of 1,2- dichloroethane,including experimental control group. low dose group(0.5ml/l),middle dose group(l.0ml/l)and high dose group(2.0ml/l),and use NMDAR and Ca²⁺antagonists respectively,which is Ketamine blocker group(2.0ml/l)and Nimodiping blocker group(2.0ml/l),every group includes±S9group,and 1,2-DCE is dissolved by Dimethyl Sulphoxide(DMSO.＜1%).(1)The cell proliferation was detected by Cell Counting Kit-â…§;(2)The cell morphological changes were observed under fluorescence microscope measured with AO-EB fluorescent staining and transmission electron microscope;(3)The expression levels of cell NMDAR and[Ca²⁺]i were determined by laser confocal microscopy;Results(1)1,2-DCE can injure the normal structural morphology of neurocyte.whatever -S9group or +S9group:the cell body swelled and brokedown,the karyon blurred or disappeared,the axone became shorten and thick,the connection among neurocytes were reduced,the cell membrane was half-baked: under transmission electron microscope we found the karyon membrane of experimental control group was whole,borderline was clear;the membrane of mitochondria was whole too,and its radges were in order,karyon membrane of 1,2-DCE groups was damaged,chromatin got together,mitochondria and endocytoplasmic reticulum showed a typical swell and deformation,the mitochondria ridges were reduced or disappeared,and the injury became severer with the dose increase of 1,2-DCE;(2)The proliferation of neurocytes in-S9group had statistically significant difference between 1,2-DCE groups and experimental control group,blocker groups(P＜0.01),but there was no statistically significant difference between experimental control group and blocker group(P＞0.05);The proliferation of neurocytes in +S9group had statistically significant difference between 1,2-DCE groups(P＜0.05),there was no statistically significant difference between low dose group and Ketamine blocker group(P＞0.05),it was the same between experimental control group and Nimodiping blocker group(P＞0.05),and the proliferation of neurocytes was reduced with the close increase of 1,2-DCE;(3)The expression of NMDARl in -S9group went up with the dose increase of 1.2-DCE in 24h administered with 1,2-DCE,and there was statistically significant difference between experimental control group and 1,2-DCE groups(P＜0.01),it is the same between 1,2-DCE groups and blocker groups(P＜0.05);(4)The[Ca²⁺]i was raised with the dose increase of 1,2-DCE in -S9groups in forepart administered with 1,2-DCE,and there was statistically significant difference among them(P＜0.01),and there was statistically significant difference between high dose group and blocker groups(P＜0.05),and the[Ca²⁺]i change was, bell-like shape with the time longer except high dose group,and the[Ca^2-]i change of high dose group was wave-like shape,and it is the same in +S9groups;Conclusion 1,2-DCE and its metabolite can damage the normal structural morphology of neurocytes,and can increase expression of NMDARI and[Ca^2-i administered with 1,2-DCE,and became severer with the dose increase of the 1.2-DCE,but the structural morphology of neurocytes in blocker groups was more better than 1,2-DCE groups.and the expression of NMDARI and[Ca²⁺]i were lower than 1,2-DCE,thus may NMDAR and Ca²⁺play an important role in acute intoxicated encephalopathy induced by 1,2-dichloroethane.