当归补血汤对小鼠脾脏衰老的分子调节

当归补血汤对小鼠脾脏衰老的分子调节(10000字)
摘  要：目的:探讨当归补血汤和对照溶液（生理盐水）对FVB种青年小鼠和衰老模型小鼠脾脏衰老的分子调节。方法:对3月龄和6月龄小鼠进行造模（第2组和第5组）与喂药（第3组和第6组）及对照（第1组和第4组），2、5组每天灌生理盐水和注射D-半乳糖，3、6组每天灌当归补血汤和注射D-半乳糖，1、4组每天灌生理盐水和注射生理盐水。45天后检测小鼠血清中超氧化物歧化酶(SOD)和小鼠溶血过氧化氢酶(CAT)水平并进行对比分析，同时取出小鼠脾脏提取总RNA并用RT-PCR方法，检测衰老相关靶基因C3和Icam1的表达。结果:当归补血汤能够提高SOD和CAT活性，当归补血汤具有清除衰老模型小鼠自由基和抑制脂质过氧化的作用。同时对基因C3和Icaml检测可知当归补血汤可能通过NF-KappaB介导的分子信号途径起作用。
关键词：当归补血汤；超氧化物歧化酶；过氧化氢酶；衰老模型

Molecular Regulation of the Mouse Spleen Aging Process by Dangguibuxuetang
Abstract: Objective: To investigate the molecular regulation mechanism of Dangguibuxuetang (DBD) and the control solution (saline) on the FVB mouse in the aging model construction process. Methods: The 3- and 6-month aged mice were used to construct aging model by injecting with D-galactose, at the same time, one of the model group was also feed with DBD, while control groups were injected with normal saline. The injection and DBD treatment lasted for 45 days. Then serum SOD and hemolysis CAT level were analyzed, at the same time, mice spleens were removed and total RNA from spleens were isolated which were used in further RT-PCR to detect age-related C3 and Icam1 gene expression levels. Results: DBD can enhance the activity of SOD and CAT, and regulate C3 and Icaml gene expression.  DBD might hinder the process of aging with clear free radicals and inhibiting lipid oxidation in mouse. While C3 and Icaml genes were known as downstream target of transcription factor NF-KappaB, DBD may play a role mediated by NF-KappaB molecular signaling pathways.
Key words: DBD; Super oxide dismutase; Catalase; Aging model
 

目    录
摘要    1
关键词    1
1 前言    2
2  材料与方法    4
2.1  材料    4
2.2  小鼠分组与给药情况    5
2.2.1  当归补血汤的配制    5
2.2.2  D-半乳糖衰老小鼠模型的建立    5
2.2.3  生理盐水的配制    6
2.3  衰老指征评定    6
2.3.1 血液中超氧化物歧化酶（SOD）活性测定    6
2.3.2  血液中过氧化氢酶（CAT）活性测定    6
2.3.3  引物设计方法    7
2.4  总RNA提取前的准备和总RNA的提取    7
2.4.1  研钵的处理    7
2.4.2  枪头及EP管的处理    8
2.4.3  从组织中提取总RNA    8
2.5  RT-PCR的原理和操作过程    8
2.5.1 RT-PCR的原理    8
2.5.2 RT-PCR的操作过程    9
3  结果与分析    10
3.1  引物设计结果    10
3.2  T-SOD活性测定结果    10
3.3  CAT活性测定结果    11
3.4  RT-PCR 鉴定各样本中所检测基因的表达水平结果    11
4  结论    13
参考文献    13
致谢    15