三七总皂甙对酒诱导兔骨髓基质干细胞成脂基因422(aP2)mRNA表达的影响.pdfVIP

目 录 中文摘要 ················································1 英文摘要 ················································3 引 言 ················································6 正 文 ···············································7 1.实验材料和方法·····································7 1.1 实验对象······································7 1.2 实验试剂·········································7 1.3 实验器材········································8 1.4 实验方法········································9 1.5 观测指标·········································10 2. 实验结果············································14 2.1 骨髓基质干细胞的生长状况 ·······················14 2.2 脂肪细胞的诱导分化和鉴定 ·······················15 2.3 三七总皂甙干预后疗效鉴定 ·······················17 2.4反转录聚合酶链式反应结果·······················18 3.讨 论······································20 4.存在的问题和展望··································32 结 论 ···············································34 参考文献 ··········································35 附 录 ···············································41 综 述 ··············································43 致 谢 ···············································49 攻读学位期间发表的学术论文目录 ····················50 作者声明 ··············································51 个人简历 ··············································52 中 文 摘 要 目 的：研究三七总皂甙能否抑制酒精诱导的骨髓基质干细胞(BMSCs) 成脂分化，及其抑制效果与剂量的关系。 方 法：从4 周龄新西兰大白兔的股骨中抽取骨髓，用密度梯度离心 法体外分离BMSCs，进行培养、传代，用倒臵相差显微镜观察细胞的形态 生长规律。将第2 代细胞传到6 孔培养板中，随机分为空白组、模型组、 高剂量治疗组、低剂量治疗组，每组10 板。空白组：在每 1ml 培养液中 加入生理盐水50μl进行诱导；模型组：在每1ml培养液中加入酒精（99.8% 无水乙醇）50μl进行诱导；高、低剂量治疗组：在每 1ml 培养液中加入 酒精50μl进行诱导成脂分化成功后，再分别加入三七总皂甙注射液100 μl、50 μl进行药物干预。对4 组细胞在药物干预前后用倒臵相差显微 镜观察形态变化，在诱导的第8 天，每组取 10 孔细胞分别行油红O 染色 脂肪细胞计数、甘油三脂含量测定、碱性磷酸酶活性测定。在干预治疗 的第15 天，每组取10 孔细胞分别行油红O 染色后脂肪细胞计数、甘油 三脂含量测定、碱性磷酸酶活性测定，成脂相关基因422 (aP )ｍRNA 表达 2 的检测。 结 果：培养的细胞在倒臵相差显微镜下观察，细