中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2005年
37期
151-153
,共3页
苯并芘类%铅%神经元%DNA损伤
苯併芘類%鉛%神經元%DNA損傷
분병비류%연%신경원%DNA손상
背景:苯并[a]芘对中枢神经和外周神经具有一定的损伤作用,苯并[a]芘与其他毒物联合神经毒作用的研究处于起始阶段.目的:应用分子生物学技术与神经元培养相结合的手段研究苯并[a]芘、铅单独及联合作用下对体外神经细胞的毒性及胞核DNA的损伤情况.设计:重复测量设计.单位:重庆医科大学劳动卫生教研室和华中科技大学同济医学院职业医学研究所热生物与分子毒理实验室.材料:实验于2003-06/09在华中科技大学同济医学院职业医学研究所热生物与分子毒理实验室完成,选择10只8日龄SD大鼠小脑组织制备原代细胞培养物,分10组培养,每组5个培养皿.按以下分组进行处理:[1]空白对照组.[2]溶剂对照组(等量二甲基亚砜+肝微粒体酶混合物平行处理).[3]低浓度铅染毒组(醋酸铅5 μmol/L).[4]高浓度铅染毒组(醋酸铅50μmol/L).[5]低浓度苯并[a]芘染毒组(苯并[a]芘5 μmol/L+肝微粒体酶混合物).[6]高浓度苯并[a]芘染毒组(苯并[a]芘50μmol/L+肝微粒体酶混合物).[7]低浓度铅+低浓度苯并[a]芘联合染毒组.[8]低浓度铅+高浓度苯并[a]芘联合染毒组.[9]高浓度铅+低浓度苯并[a]芘联合染毒组.[10]高浓度铅+高浓度苯并[a]芘联合染毒组.方法:染毒90min,胰酶消化法收集标本,椎虫蓝染色法检测各组细胞存活率;单细胞凝胶电泳法检测各组细胞胞核DNA的损伤情况.主要观察指标:苯并[a]芘、铅单独及联合染毒下体外神经细胞的存活率及胞核DNA的损伤率. 结果:[1]两种浓度的苯并[a]芘、铅单独或联合染毒各组细胞存活率均低于对照组[染毒组为(44.14±4.80)%～(82.40±2.70)%,对照组为(88.44±2.53)%～(90.12±2.23)%,P＜0.05～0.01].[2]高浓度苯并[a]芘单独染毒组、低浓度铅+高浓度苯并[a]芘染毒组、高浓度铅+低浓度苯并[a]芘染毒组、高浓度铅+高浓度苯并[a]芘染毒组胞核DNA损伤程度均高于对照组[63%(19/30),87%(26/30),80%(24/30),97%(29/30),13%(4/30),20%(6/30),P＜0.01].结论:苯并[a]芘与铅均有一定的体外神经毒性,且二者有协同作用;苯并[a]芘损害体外培养神经元胞核DNA的能力大于铅.
揹景:苯併[a]芘對中樞神經和外週神經具有一定的損傷作用,苯併[a]芘與其他毒物聯閤神經毒作用的研究處于起始階段.目的:應用分子生物學技術與神經元培養相結閤的手段研究苯併[a]芘、鉛單獨及聯閤作用下對體外神經細胞的毒性及胞覈DNA的損傷情況.設計:重複測量設計.單位:重慶醫科大學勞動衛生教研室和華中科技大學同濟醫學院職業醫學研究所熱生物與分子毒理實驗室.材料:實驗于2003-06/09在華中科技大學同濟醫學院職業醫學研究所熱生物與分子毒理實驗室完成,選擇10隻8日齡SD大鼠小腦組織製備原代細胞培養物,分10組培養,每組5箇培養皿.按以下分組進行處理:[1]空白對照組.[2]溶劑對照組(等量二甲基亞砜+肝微粒體酶混閤物平行處理).[3]低濃度鉛染毒組(醋痠鉛5 μmol/L).[4]高濃度鉛染毒組(醋痠鉛50μmol/L).[5]低濃度苯併[a]芘染毒組(苯併[a]芘5 μmol/L+肝微粒體酶混閤物).[6]高濃度苯併[a]芘染毒組(苯併[a]芘50μmol/L+肝微粒體酶混閤物).[7]低濃度鉛+低濃度苯併[a]芘聯閤染毒組.[8]低濃度鉛+高濃度苯併[a]芘聯閤染毒組.[9]高濃度鉛+低濃度苯併[a]芘聯閤染毒組.[10]高濃度鉛+高濃度苯併[a]芘聯閤染毒組.方法:染毒90min,胰酶消化法收集標本,椎蟲藍染色法檢測各組細胞存活率;單細胞凝膠電泳法檢測各組細胞胞覈DNA的損傷情況.主要觀察指標:苯併[a]芘、鉛單獨及聯閤染毒下體外神經細胞的存活率及胞覈DNA的損傷率. 結果:[1]兩種濃度的苯併[a]芘、鉛單獨或聯閤染毒各組細胞存活率均低于對照組[染毒組為(44.14±4.80)%～(82.40±2.70)%,對照組為(88.44±2.53)%～(90.12±2.23)%,P＜0.05～0.01].[2]高濃度苯併[a]芘單獨染毒組、低濃度鉛+高濃度苯併[a]芘染毒組、高濃度鉛+低濃度苯併[a]芘染毒組、高濃度鉛+高濃度苯併[a]芘染毒組胞覈DNA損傷程度均高于對照組[63%(19/30),87%(26/30),80%(24/30),97%(29/30),13%(4/30),20%(6/30),P＜0.01].結論:苯併[a]芘與鉛均有一定的體外神經毒性,且二者有協同作用;苯併[a]芘損害體外培養神經元胞覈DNA的能力大于鉛.
배경:분병[a]비대중추신경화외주신경구유일정적손상작용,분병[a]비여기타독물연합신경독작용적연구처우기시계단.목적:응용분자생물학기술여신경원배양상결합적수단연구분병[a]비、연단독급연합작용하대체외신경세포적독성급포핵DNA적손상정황.설계:중복측량설계.단위:중경의과대학노동위생교연실화화중과기대학동제의학원직업의학연구소열생물여분자독리실험실.재료:실험우2003-06/09재화중과기대학동제의학원직업의학연구소열생물여분자독리실험실완성,선택10지8일령SD대서소뇌조직제비원대세포배양물,분10조배양,매조5개배양명.안이하분조진행처리:[1]공백대조조.[2]용제대조조(등량이갑기아풍+간미립체매혼합물평행처리).[3]저농도연염독조(작산연5 μmol/L).[4]고농도연염독조(작산연50μmol/L).[5]저농도분병[a]비염독조(분병[a]비5 μmol/L+간미립체매혼합물).[6]고농도분병[a]비염독조(분병[a]비50μmol/L+간미립체매혼합물).[7]저농도연+저농도분병[a]비연합염독조.[8]저농도연+고농도분병[a]비연합염독조.[9]고농도연+저농도분병[a]비연합염독조.[10]고농도연+고농도분병[a]비연합염독조.방법:염독90min,이매소화법수집표본,추충람염색법검측각조세포존활솔;단세포응효전영법검측각조세포포핵DNA적손상정황.주요관찰지표:분병[a]비、연단독급연합염독하체외신경세포적존활솔급포핵DNA적손상솔. 결과:[1]량충농도적분병[a]비、연단독혹연합염독각조세포존활솔균저우대조조[염독조위(44.14±4.80)%～(82.40±2.70)%,대조조위(88.44±2.53)%～(90.12±2.23)%,P＜0.05～0.01].[2]고농도분병[a]비단독염독조、저농도연+고농도분병[a]비염독조、고농도연+저농도분병[a]비염독조、고농도연+고농도분병[a]비염독조포핵DNA손상정도균고우대조조[63%(19/30),87%(26/30),80%(24/30),97%(29/30),13%(4/30),20%(6/30),P＜0.01].결론:분병[a]비여연균유일정적체외신경독성,차이자유협동작용;분병[a]비손해체외배양신경원포핵DNA적능력대우연.
BACKGROUND: Benzo[a]pyrene injures central and peripheral nerves at a certain degree. The study is at an initial phase concerning to the neurotoxicity of benzo[a]pyrene allied with other toxic objects.OBJECTIVE: Molecular biological technique integrated with neuron culture were applied in the study of benzo[a]pyrene and lead applied separately or in combination on neurotoxicity and nuclear DNA damage in vitro.DESIGN: Repeated measure.SETTING: Teaching-Research Room of Labor and Hygiene of Chongqing Medical University and Laboratory of Thermobiology and Molecular Toxicology of Occupation Medical Institute of Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Laboratory of Thermobiology and Molecular Toxicology of Occupation Medical Institute of Tongji Medical College ofHuazhong University of Science and Technology from June to September 2003. Ten SD rats of 8-day old were employed and their brain tissues were prepared as primary cell culture object, which were divided into 10 culture groups, 5 culture dishes in each group. The managements were as follows in each group: [1] blank control; [2] solvent control[parallel management with equivalent dimethyl sulfoxide (DMSO) +liver microsome enzyme]; [3] lead group of low concentration (acetic lead 5 μmol/L) (No. 1 group); [4] lead group of high concentration (acetic lead 50 μmol/L) (No. 2 group); [5] benzo[a]pyrene group of low concentration (benzo[a]pyrene 5 μmol/L + liver microsome enzym) (No. 3 group); [6]benzo[a]pyrene group of high concentration (benzo[a]pyrene 50 μmol/L +liver microsome enzym) (No. 4 group); [7] lead of low concentration +benzo[a]pyrene of low concentration group (No. 5 group); [8] lead of low concentration + benzo[a]pyrene of high concentration group (No. 6 group);[9] lead of high concentration + benzo[a]pyrene of low concentration group (No.7 group); [10] lead of high concentration + benzo[a]pyrene of high concentration group (No. 8 group).METHODS: After stained poisoning for 90 minutes, trypsin digestion method was used for sample collection and trypan blue staining method was applied to assay cell survival rate in each group. Single-cell gel electrophoresis (SCGE) was used to determine the damage of nuclear DNA in each group.MAIN OUTCOME MEASURES: Neuronal survival rate and damage rate of nuclear DNA in poisoning with benzo[a]pyrene and lead either in separation or in combination.RESULTS: [1] Cell survival rates in various groups poisoned with benzo [a]pyrene and lead of two concentrations either in separation or combination were lower than the controls [poisoning group (44.14±4.80)% to(82.40±2.70)%, the controls (88.44±2.53)% to ( 90.12±2.23)%, P ＜ 0.05to 0.01]. [2] Degrees of nuclear DNA damage in single poisoning group with benzo[a]pyrene of high concentration, lead of low concentration + benzo[a]pyrene of high concentration, lead of high concentration + benzo[a]-pyrene of low concentration and lead of high concentration + benzo[a]pyrene of high concentration were higher than the controls [63% (19/30), 87%(26/30), 80% (24/30), 97% (29/30), 13% (4/30), 20% (6/30), P ＜ 0.01].CONCLUSION: Both benzo[a]pyrene and lead present neurotoxicity in vitro and coordinate with each other. The damage of benzo[a]pyrene is worse than lead in neuronal nuclear DNA cultured in vitro.
