毕业论文--山茱萸生品及制品二氯甲烷萃取部位对D-半乳糖致衰小鼠骨髓细胞的影响.doc

山茱萸生品及制品二氯甲烷萃取部位对D-半乳糖致衰小鼠骨髓细胞的影响 作者：赵青春,董菊,杨柳,王明艳,杜逸芳 【摘要】 目的 研究山茱萸生、制品二氯甲烷萃取部位对D-半乳糖致衰小鼠骨髓细胞的影响。方法 单细胞电泳观察D-半乳糖致衰小鼠骨髓细胞DNA损伤情况,RT-PCR法检测小鼠骨髓细胞p53基因的表达情况。结果 D-半乳糖致衰模型组小鼠骨髓细胞DNA拖尾率高于正常对照组(P＜0.05),且Ⅲ、Ⅳ级(中重度)损伤细胞数较多；山茱萸生品及制品二氯甲烷萃取部位组小鼠骨髓细胞DNA的拖尾率均低于D-半乳糖致衰模型组(P＜0.05)。D-半乳糖致衰模型组小鼠骨髓细胞p53基因的表达高于正常对照组、阳性药组及制品组。结论 D-半乳糖致衰小鼠骨髓细胞DNA有损伤,山茱萸生、制品二氯甲烷萃取部位对D-半乳糖致衰小鼠骨髓细胞有保护作用；D-半乳糖致衰小鼠骨髓p53基因表达上调,山茱萸制品二氯甲烷萃取部位能显著下调D-半乳糖致衰小鼠骨髓p53基因的表达。 【关键词】 山茱萸；二氯甲烷萃取部位；D-半乳糖；骨髓细胞；小鼠 　　Abstract：Objective To study the influences of dichloromethane extraction from crude and processed Cornus Officinalis on bone marrow cells of aging mice induced by D-galactose. Methods DNA damage of bone marrow cells of aging mice induced by D-galactose was detected by the comet assay, the expression of p53 mRNA was detected by RT-PCR. Results The tail formation rate of model group was significantly higher than the rate of control group (P＜0.05) with a large number of Ⅲ and Ⅳ DNA damage cells. The tail formation rate of dichloromethane extraction groups were both significantly lower than the rate of model group (P＜0.05). p53 mRNA expression of model group was higher than the rest except the crude Cornus Officinalis group. Conclusion There is DNA damage of bone marrow cells of aging mice induced by D-galactose. Both dichloromethane extraction from crude and processed Cornus Officinalis are protective. p53 mRNA expression up-regulates in model group, while its significantly down-regulation is detected in the processed Cornus Officinalis group. 　　Key words：Cornus Officinalis；dichloromethane extraction；D-galactose；bone marrow cells；mice 　　山茱萸为山茱萸科落叶小乔木植物山茱萸(Cornus officinalis Sieb.et Zucc.)的干燥成熟果肉,又名枣皮、山芋、药枣、肉枣(《本草纲目》)、实枣儿(《救荒本草》)等[1-2],其味酸、涩,微温,归肝、肾经,功能补益肝肾、涩精固脱。临床用于内热消渴、眩晕耳鸣、腰膝酸痛、阳痿遗精、尿遗尿频、崩漏带下、大汗虚脱等。近年研究表明,山茱萸具有调节免疫、降血糖、抗休克、抑制血小板聚集、抗氧化、抗炎、抗菌、抗癌、保肝、治疗不育症等多种药理作用[3]。本实验在以往课题研究的基础上,选取山茱萸生、制品二氯甲烷萃取部位,研究其对D-半乳糖致衰小鼠骨髓细胞的影响。 　　1 实验材料 　　1.1 药物与试剂 　　吐温-80、D-半乳糖,国药集团化学试剂有限公司,批号生理盐水,上海百特医疗用品有限公司；维生素E胶丸,浙江医药股份有限