過敏性氣喘中醫熱證的基因表現

GENE EXPRESSION IN “HEAT” ZHENG PATTERN OF ALLERGIC ASTHMA

本研究利用微陣列生物晶片分析過敏性氣喘基因表現的差異與中醫哮喘熱證之間的相關性，以了解哮喘熱證表現背後的實質分子機轉。本研究收集95 位合乎過敏性氣喘臨床診斷標準的病人，並根據中醫傳統理論對各種臨床症狀發生頻率與嚴重程度加以評分，作為熱證程度的評量。同時，採取病人週邊血中的白血球利用微陣列生物晶片的技術，分析其基因表現的變化。以無母數相關分析法（Spearmancorrelation ）分析各基因的表現量與熱證評分間的相關性。結果發現Thromboxan A2 receptor （TBXA2R ）（p = 0.010 ）、CD2 （p = 0.043 ）、Interleukin-15 （IL-15 ）（p = 0.043 ）、Regulated uponActivation, Normal T cell Expressed and Secreted （RANTES ）（p < 0.001 ）、SCYA4 （MIP-1b ：macrophageinflammatory protein-1b ）（p = 0.009 ）、SCYA17 （TARC ：Thymus and activation-related chemokine ）（p = 0.002 ），這些基因與熱證評分呈正相關，並具有統計上意義或顯著意義。將上述基因表現量與熱證評分進行多變項迴歸分析（Multiples Linear Regression ），得出一回歸方程式：Heat = 0.49 （TBXA2R ）+ 1.23 （Eotaxin ）+ 0.93 （STAT2 ）+ 5.00 （GBP2 ）+ 0.88 （Metallothionein ）- 3.25 （CD2 ）+ 2.24 （IL-15 ）- 12.59 （IL-18 ）+ 6.50 （RANTES ）- 2.24 （SCYA4 ）+ 1.19 （SCYA17 ）該模式具顯著意義（p < 0.05 ），且可預測熱證評分準確性達70.6 % 。從上述結果中，我們發現哮喘熱證趨化激素如RANTES 、SCYA4 、SCYA17 有最強的相關性，同時也與訊息傳導分子TBXA2R 、細胞激素IL-15 及IL-18 、黏附分子CD2 成正相關，這些基因必定在氣喘熱證的病理機轉中扮演重要角色。同時，這些基因與熱證評分進行多變項迴歸所得到的迴歸方程式，可以預測病人熱證的評分，以作為臨床判斷過敏性氣喘熱證程度方便且快速的客觀標準，也可用於治療的評估。The pathogenesis of allergic asthma involves a very complicate immune response in which thousands of genes are expressed. “Zheng” is an unique theory of both diagnosis and treatment in traditional Chinese medicine (TCM). In this study, we try to analysis the correlation between heat “Zheng” score of allergic asthma patients and the differences of their gene expression in order to understand the molecular mechanism of heat “Zheng” in asthma. Therefore, we included 95 allergic asthma patients in our study. According to the theory of TCM, we decided their heat “Zheng” score via the frequency and severity of their clinical symptoms. At the same time, their peripheral monocytes were collected. So, we used microarray to detect the gene expression of patients’ peripheral lymphocytes which are involved in the pathogenesis of allergic asthma. Then, the results were analyzed by Spearman correlation statistics. We found Thromboxan A2 receptor (TBXA2R) (p = 0.010)、CD2 (p = 0.043)、Interleukin-15 (IL-15) (p = 0.043)、Regulated upon Activation, Normal T cell expressed and secreted (RANTES) (p < 0.001)、SCYA4 (MIP-1b：macrophage inflammatory protein-1b) (p = 0.009)、SCYA17 (TARC：Thymus and activation-related chemokine) (p = 0.002)， were positive correlated to heat “Zheng” score which were significant in statistics (p < 0.05). A regression analysis was then performed. We obtained a formulation: Heat = 0.49 (TBXA2R) + 1.23 (Eotaxin) + 0.93 (STAT2) + 5.00 (GBP2) + 0.88 (Metallothionein) – 3.25 (CD2) + 2.24 (IL15) – 12.59 (IL18) + 6.50 (RANTES) –2.24 (SCYA4) + 1.19 (SCYA17), which allowed us to predict patient’s heat “Zheng” score by the expression amount of those genes. The accuracy is 70.6 %. On the base of the results, we can conclude that signaling factor TBXA2R, cytokines IL-15 and IL-18, adhesion molecular CD2 and chemokins RANTES, SCYLA4, SCYA17 must play important roles in allergic heat “Zheng” of asthma pathogenesis. The regression formulation we obtained has great value of clinical application. It provides an objective evidence to diagnose allergic heat “Zheng” asthma and to estimate the efficacy of therapy.