红花注射液对气虚血瘀证模型大鼠基因调控作用的研究

目的:采用基因芯片技术,分别构建气虚血瘀证大鼠和红花注射液给药处理后气虚血瘀证大鼠的差异基因表达谱,比较并分析,筛选出红花能够治疗气虚血瘀证的关键基因群,并推测其起治疗作用的基因组调控机制。方法:15只SD大鼠随机分为模型组、给药组、空白对照组。模型组和给药组采用疲劳游泳和饥饿饲养处理。造模一周后,给药组尾静脉注射红花注射液(100mg/kg/d),模型组给予相同体积生理盐水;对照组不做任何处理。造模进行两周后处死大鼠,取血检验血流变指标并评价造模情况;另抽取足够的血分离mRNA并逆转录杂交基因芯片;扫描信号分析确定受红花注射液调控的基因;并通过基因数据库查询相关基因功能,结合相关文献分析初步探讨红花作用的机制。结果:两周后经过检验和观察发现模型组大鼠在不同切率下的全血粘度增加,并且其体征表现出虚弱和瘀血的状态、体重下降,确定造模成功;给药组大鼠则相对于模型组的各项检测指标和状态有所改善,确认药物有疗效。在差异基因的比较中,空白组相对于给药组上调基因252条,下调基因54条;给药组相对于模型组上调基因196条,下调基因32条;两次差异表达基因中有16条相同基因,这些差异基因涉及到炎症损伤、免疫调节反应等方面。结论:红花注射液对于气虚血瘀证有治疗作用,在基因层次上是通过抗炎症损伤机制实现的。

Study on the Effect of Safflower Injection on the Regulation of Genes related to Qi-deficiency and Blood Stasis Syndrome in Rats

ABSTRACT Objective: To establish two differential gene expression profiles of qi-deficiency and blood stasis syndrome before or after safflower injection treatment by using gene chip technology; compared and analyzed to ensure the effective genes that are responsible for the therapeutic effects of safflower injection against qi-deficiency and blood stasis syndrome in rats. Furthermore, speculated the effect mechanism of the therapeutic genes. Methods: Fifteen SD rats were randomly divided into 3 groups (n=5): control group, model group, and medication group. Qi-deficiency and blood stasis model was established by subjecting the rats to hunger and fatigue for two weeks. After a week of the modeling, safflower injection (100 mg/kg/d) was administered daily via the tail vein for 7 days in medication group, and the rats in model group were injected with saline of the same volume. Control group received normal feeding. At the end of the experiment, rats were killed and whole blood was collected to evaluate the blood stream change and extract mRNAs in blood samples. Qualified mRNAs were reverse transcribed into cDNA which was then used in gene chip hybridization. The genes regulated by safflower injection were determined by the fluorescence signal and the functional mechanisms of safflower injection were confirmed by further querying genealogy databases and reviewing literatures. Results: After two weeks of the modeling, the whole blood viscosity under various shear rates was significantly increased in the model rats which showed faint, blood stasis and weight loss, indicating that the model is made successfully. The increased whole blood viscosity and qi-deficiency and blood stasis syndrome were obviously reversed by safflower injection treatment. Compared with the control group, 252 genes up-regulated while 54 genes down-regulated in model group; compared with the model group, 196 genes up-regulated while 32 genes down-regulated. Among these, 16 differentially expressed genes were involved in inflammation and immune response. Conclusion: Safflower injection was effective in treating qi deficiency and blood stasis syndrome, which was achieved by regulating inflammation related genes.