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1、甲醛和二甲苯联合染毒对小鼠骨髓细胞遗传毒性研究 作者： 日期：甲醛和二甲苯联合染毒对小鼠骨髓细胞遗传毒性研究 基金工程 山西医科大学科技创新基金资助工程01200720作者单位 山西医科大学公共卫生学院环境卫生学教研室山西 太原 030001作者简介 白雪1986-，女，硕士研究生； 研究方向：环境卫生学；E-mail：baixue30405126 *通讯作者 原福胜 E-mail：fsyuansohu 白雪，原福胜*，张婷，王君霞，王鸿摘要：目的 探讨甲醛和二甲苯联合腹腔注射染毒对小鼠骨髓细胞的遗传毒性作用。方法 安康清洁级昆明种小鼠78只，雌雄各半，体重1822g，按体重将小鼠随机分为13
2、组，每组6只。分别为甲醛低(5mg/kg)、中(10mg/kg)、高(20mg/kg)剂量组以及生理盐水0.01ml/g阴性对照组；二甲苯低(50mg/kg)、中(100mg/kg)、高(150mg/kg)剂量组以及花生油0.01ml/g阴性对照组；联合染毒低(2.5mg/kg甲醛+25mg/kg二甲苯)、中(5mg/kg甲醛+50mg/kg二甲苯)、高(10mg/kg甲醛+75mg/kg二甲苯)剂量组以及生理盐水0.005ml/g+花生油0.005ml/g阴性对照组；微核试验采用环磷酰胺40 mg/kg作为阳性对照组。采用腹腔注射染毒，每天一次，连续7天。染毒完毕次日处死小鼠，采用微核试验和
3、彗星试验检测甲醛和二甲苯联合染毒致骨髓细胞的遗传毒性作用。结果 甲醛以及二甲苯均可引起小鼠骨髓细胞微核率增高P0.05，且随着染毒剂量增加微核率上升，具有较好的剂量反响关系；甲醛与二甲苯联合染毒微核率高于分别单独染毒，在联合染毒高剂量组时尤其明显P0.05。甲醛和二甲苯均可使小鼠骨髓细胞DNA发生损伤作用且在高剂量组损伤明显，二者联合染毒在较低剂量时便致彗星细胞尾部DNA含量及尾矩形成大于甲醛和二甲苯单独染毒，随着染毒剂量的增高联合染毒毒性作用明显增高P0.05。结论 甲醛、二甲苯对小鼠骨髓细胞的微核率和彗星细胞尾部DNA含量以及尾矩均有一定的损伤作用，二者联合染毒可能存在协同毒性效应。关键词
4、：甲醛；二甲苯；骨髓细胞；微核试验；彗星试验Study of Joint Toxic Effect of Formaldehyde and Xylene on Genetic Toxicity of Mouse Bone Marrow CellsBaixue, Yuan Fusheng*, Zhangting, Wang Junxia, WanghongAbstract: Objective Exploretheeffectofjointintraperitonealinjectionof formal-dehydeandxyleneonthegenetic toxicityofmousebo
5、nemarrowcells. Methods 78 healthy and clean Kunming mice are selected, with an equal number of males and females. The weight of each mouse is 1822g. The mice randomly are divided into 13 groups according to the weight, with 6 mice in each group. It can be divided into as follows: the group with a lo
6、w dose of formaldehyde (5mg/kg), that with a middle dose (10mg/kg), and that with a high dose (20mg/kg) as well as a negative control group with normal saline (0.01ml/g); the group with a low dose of xylene (50mg/kg), that with a middle dose (100mg/kg), and that with a high dose (150mg/kg) as well a
7、s a negative control group with peanut oil (0.01ml/g); the group with a low dose of joint exposure (2.5mg/kg formaldehyde +25mg/kg xylene), that with a middle dose (5mg/kg formaldehyde +50mg/kg xylene), and that with a high dose (10mg/kg formaldehyde +75mg/kg xylene) as well as a negative control gr
8、oup with normal saline (0.005ml/g) + peanut oil (0.005ml/g); The micronucleus test uses cyclophosphamide (40mg/kg) as a positive control group.The mice were exposed to formaldehyde and xylene by peritoneal injection for 7 days,once per day.After the mice are infected, they will be killed in the foll
9、owing day. The micronucleus test and comet assay are carried out to detect the joint toxic effect of formaldehyde and xylene on the genetic toxicity of mouse bone marrow cells. ResultsFormaldehyde and xylene can increase the micronucleus rate of mouse bone marrow cells (P0.05). With the increase of
10、exposure dosage, the micronucleus rate increases, which shows a good dose-response relationship; the micronucleus rate of joint exposure of xylene and formaldehyde is higher than that of independent exposure, which is especially obvious in the group with high dosage of joint exposure (P0.05). Xylene
11、 and formaldehyde will cause DNA damages in mouse bone marrow cells and the damages are obvious in the group with a high dosage. The joint exposure at a low dosage can cause larger tail DNA content of the comet cells and tail moment than the independent exposure of formaldehyde or xylene. With the i
12、ncrease of exposure dosage, the joint toxic effect significantly increases (P0.05).Conclusions Formaldehyde and xylene have a certain effect on micronucleus rate of mouse bone marrow cells and the tail DNA content of the comet cells and tail moment, and the joint exposure may have a synergistic toxi
13、c effect.Keywords: formaldehyde; xylene; bone marrow cell; micronucleus test; comet assay甲醛和二甲苯是我国室内环境的主要污染物，二者不仅污染水平高，而且生物毒性较高，其对安康的危害日益受到普遍关注。据报道，甲醛会对神经系统、免疫系统、生殖系统造成危害，并具有遗传毒性，长期接触一定浓度甲醛会对暴露者的血液、组织及细胞产生遗传毒性作用1-2。二甲苯具有一定的神经毒性、肾毒性和胚胎毒性3-4。国内外对二者联合遗传毒性效应研究较少。本研究拟通过甲醛和二甲苯联合染毒，采用微核试验及彗星试验探讨甲醛与二甲苯联合染毒对
14、小鼠骨髓细胞的遗传毒性。1. 材料与方法1.1 仪器与试剂 DYCP-33A 电泳槽北京市六一仪器厂，DYY-7C 电泳仪北京市六一仪器厂，光学显微镜SX212南京江南永新光学，BX51 荧光显微镜日本OLYMPUS公司。 甲醛分析纯，天津市天力化学试剂；二甲苯分析纯，天津市星月化工；甲醇天津市富宇精细化工，Giemsa粉分析纯，上海试剂三厂正常熔点琼脂糖NMA，低熔点琼脂糖LMA，Amresco公司，二甲基亚砜天津市化学试剂一厂，Triton-x-100Amresco公司，十二烷基肌氨酸钠SLS，Amresco公司，三羟甲基氨基甲烷Tris，Sigma公司，乙二胺四乙酸二钠盐Na2-EDTA
15、，北京化工厂，溴化乙锭EB，Amresco公司，0.4%台盼蓝溶液Sigma公司，其他试剂均为国产分析纯。其它试剂均为国产分析纯。1.2 实验动物安康清洁级昆明种小鼠78只，雌雄各半，体重1822g，由山西医科大学动物中心提供许可证号：SCXK(晋)20210001； 动物房合格证书号：医动字070102号。1.3 方法1.3.1 动物分组和染毒按体重将小鼠随机分为13组，每组6只。采用腹腔注射染毒，每天1次，连续7天。各组染毒剂量根据小鼠腹腔注射甲醛和二甲苯的LD50的文献资料而定5-6，分别为甲醛低(5mg/kg)、中(10mg/kg)、高(20mg/kg)剂量组以及生理盐水0.01ml/g阴性对照组；二甲苯低(50mg/kg)、中(100mg/kg)、高(150mg/kg)剂量组以及花生油0.01ml/g阴性对照组；联合染毒低(2.5mg/kg甲醛+25mg/kg二甲苯)、中(5mg/kg甲醛+50mg/kg二甲苯)、高(10mg/kg甲醛+75mg/kg二甲苯)剂量组以及
