基因芯片检测灵芝提取物对白细胞中免疫相关基因表达的影响

Influence of Lingzhi extract on immunity-related gene expression in leucocytes detected by gene chip technology

目的:灵芝对于免疫系统的调节还缺乏严格的临床证据,其关键是缺乏客观、系统评价灵芝对免疫系统调节效果的手段。基于此,采用基因芯片分析检测了灵芝对免疫相关基因表达水平的影响,从而揭示与灵芝功能密切相关的基因和途径。方法:实验于2002-12/2003-05在首都医科大学宣武医院神经分子生物实验室完成。①实验干预:血液材料来源于身体健康的宣武医院医护人员和学生12人,均对检测项目知情同意。平均年龄(31±4)岁,男女各6人。将其分为年龄、性别大致匹配的2组:安慰剂组和灵芝组,每组6人。灵芝组和安慰剂组对象分别服用灵芝胶囊和淀粉胶囊安慰剂,每人每天服用2次,每次2粒(相当于药用灵芝400mg),连续30d。灵芝胶囊主要成分为软壁灵芝孢子粉、灵芝提取物。②实验评估:服药前1天和服药30d后采用自动分析仪计算全血不同种类白细胞计数和不同种类白细胞所占比例。分离白细胞抽提总RNA,利用博星公司的cDNA芯片Biostar-H-IC-I检测了服用灵芝胶囊后白细胞中441个免疫相关基因的表达变化。结果:①所检测的441个免疫相关基因中,共有11个基因存在着差异表达。其中10个基因表达水平升高,1个基因表达水平降低。发生差异表达的11个基因中有2个参与信号转导过程,包括STAT4和LYN。服用灵芝胶囊后两个基因的表达量都显著增加。在发生差异表达的11个基因中有7个基因属于淋巴细胞表面抗原。②服用灵芝胶囊和安慰剂后白细胞总数和不同种类白细胞的百分比均未发生明显变化(P>0.05)。结论:①灵芝对免疫系统的调节功能主要通过调节淋巴细胞的活性而实现,而不是通过改变白细胞数量和分类计数。②STAT4和LYN参与的信号转导途径很有可能是灵芝胶囊作用的主要靶点。

Lacking of the objective and systematic methods has proved to be the main obstacles of the related clinical evidences to evaluate the effects of Lingzhi on immune system. Therefore, microarray analysis was used to examine the effects of Lingzhi on gene expression profile and figure out the genes and pathway activated by Lingzhi. METJPDS：The expedment was carded out in Laboratory of Neural Molecular Biology, Xuanwu Hospital, Capital Medical University from December 2002 to May 2003. ①The whole blood was collected from 12 healthy medical staffs and students in Xuanwu Hospital after providing informed consent. These volunteers, including 6 males and 6 females, with average age of （31±4） years ware divided into two age-and-sex matched groups which ware given Lingzhiand placebo capsules, respectively with 6 in each group. Capsules ware taken twice a day, and two pieces per time for 30 days, equaling to 400 mg of pure Lingzhi were taken per day. The main ingredients of Lingzhi capsule are Lingzhi extract＇and spores powder. ②The whole blood ware collected one day before or 30 days after drug administration. Total number and proportion of each leucocytes ware analyzed using auto-analyzer. Total RNA was extracted from leucocytes. Expression profiles of 441 immune-related genes were analyzed with gene chips Biostar-H-IC-I produced by Boxing Company to figure out the differently expressed genes induced by Lingzhiadministretion. RESULTS： ①Eleven out of 441 genes differently expressed in response to Lingzhi induction, with 10 elevated and 1 decreased, among which, two key regulators of signal transduction, e.g. STAT4 and LYN ware activated significantly by LingzhL In addition, 7 differently expressed genes ware surface receptors of lymphocytes. ② No significant differences ware observed on the total number and percentage of leucocytes after taking Lingzhi and placebo （P 〉 0.05）. CONCLUSION： ①Lingzhi can regulate lymphocyte activity by modulating gene expression profile, but not alter the leucocytes number or the proportion. ②lt is most likely that STAT4 and LYN related signal transduction pathways can mediate the Lingzhi induced regulation.