用基因芯片筛选针刺衰老大鼠涌泉穴的318个差异基因初报

Preliminary Analysis of 318 Differentially Expressed Genes Screened out by cDNA Microarray of Aging Rat Model Treated by Acupunctured at Yongquan Point

目的 :应用基因芯片研究针刺涌泉穴对D 半乳糖所致衰老大鼠基因表达的影响。方法 :将成年健康大鼠随机分为模型组与针刺组 ,两组每日皮下注射 10 %D 半乳糖 0 5ml造模 ,造模的同时针刺组予以针刺涌泉穴 ,7周后处死 ,并摘取肝脏提取mRNA ,作逆转录为cDNA以备检测。随机选用 2 30 5条大鼠cDNA制备成表达谱芯片 ,用Cy3和Cy5两种荧光物质分别标记两组大鼠肝组织的cDNA ,制备成探针与表达谱芯片进行杂交 ,通过计算机扫描后进行数据分析 ,初步筛选出了差异表达基因 318条 ,其中表达上调的有 137条 ,表达下调的有 181条 ,并对其中差异最为显著的 5 5条作了初步分析。结果 :针刺涌泉穴对衰老大鼠的基因表达的影响涉及免疫、生长发育、基础代谢和细胞骨架运动等多个方面 ,这从基因水平证明了针刺治疗是一整体调节作用。结论 :应用基因芯片技术能够同时半定量的研究数千条基因的表达状态 ,可以从微观基因水平研究针刺的整体机理 ,并判定其分子疗效 。

Objective: To study the effect of acupuncture Yongquan point on gene expression in aging rat model induced by D galactose using cDNA microarrays. Methods: Rats were divided to the model group and the acupuncture group at random. Aging model were made by subcutaneous injection of D galactose. The model rats in the acupuncture group were treated by acupunctured at Yongquan point every day for 5 times a week. The rats were sacrificed after seven weeks, the liver was taken out to extract mRNA for reverse transcription cDNA. 2305 cDNA were elected to made microarray for analysis. cDNA extracted from model rats were labeled with Cy3 dUTP while cDNA extracted from acupuncture group were labeled with Cy5 dUTP. These probes were hybridized with cDNA microarray, scanned and analyzed. There were 318 differentially expressed in rats of the acupuncture group, 137 were up regulated and 181 were down regulated. Priliminary analysis was made on 55 cDNA with most significant difference. Results: Acupuncture has vast effects on aging rats′ gene expression, involving multiple scopes as immunity, growth and development, basal metabolism and cell skeleton motion. Acupuncture is proved, on gene level, to have a holistic regulation. Conclusion: The state of thousands gene expression could be studied simultaneously by cDNA microarray semiquantitatively, through which the holistic mechanism of acupuncture could be investigated on microcosmic gene level to evaluate its molecular effect, so as to raise the level of TCM research.