肺气虚证患者T淋巴细胞相关基因的表达研究

Related Gene Expression of Thymus Dependent Lymphocyte in Patient with Syndrome of Deficiency of Lung-qi

目的:通过基因芯片技术研究肺气虚证患者T淋巴细胞基因表达的差异。方法:采集肺气虚证和肺阴虚证患者、正常人的外周血作为实验血样和对照血样。采用Ficoll技术分离外周血淋巴细胞、流式细胞仪分选并纯化T淋巴细胞,一步法提取总RNA,逆转录合成双链cDNA,体外转录合成生物素标记的cRNA,片断化后,采用人类全基因表达谱芯片进行芯片杂交,扫描后筛选出差异表达基因。结果:肺气虚证患者/正常人外周血T淋巴细胞相关差异表达基因45条,其中上调41条,下调4条;肺气虚证/肺阴虚证患者外周血T淋巴细胞相关差异表达基因43条,其中上调27条,下调16条;肺气虚证组/肺阴虚证组、正常人组均高表达的差异基因15条。结论:基因芯片技术能有效地研究中医证的基因表达谱,筛查出肺气虚证患者T淋巴细胞相关差异表达基因。

Objective： To study the related gene expression of T lymphocyte in the patient with syndrome of deficiency of lung-qi （SDLQ） by gene chip. Methods： Collected peripheral blood of patients with SDLQ （as experimental samples） and patients with pulmonary yin deficiency syndrome （PYDS） and normal men （as control samples）, By Ficoll method, peripheral blood lymphocytes were collected, then thymus dependent lymphocytes were extracted and purified by flow cytometry. Total RNA were extracted by one-step technique and purified, then were synthesized to double strand cDNA templates by reverse transcription, and synthesized to cRNA probes with biotin labeling in vitro transcription. Subsequently cRNA samples were fragmented. The gene chip was hybridized and scanned. Then fluorescent signal value of gene expression was obtained, and differential expression genes were sifted. Results： There were 45 related differential expression genes in T lymphocytes of SDLQ patients and normal men, including up-regulated 41 and down-regulated 4; 43 related differential expression genes in T lymphocytes of SDLQ and PYDS patients, including up-regulated 27 and down-regulated 16. There were 15 highly expressed differential genes in all SDLQ group, PYDS group and normal group. Conclusion： Gene chip can be applied to study gene expression profiles of syndrome in TCM effectively and to screen the related gene expression of T lymphocyte in SDLQ patient.