寒熱證型小兒氣喘患者基因表現之研究 研究生：魏芳慧 指導教授:高尚德 教授 中國醫藥大學中國醫學研究所 過敏性氣喘是兒科臨床常見的疾病，推測可能受基因（genetic factors）和環境刺激相互作用。中醫兒科認為氣喘發作病變在肺，證分寒熱；本研究欲了解其基因表現之差異。本研究設計《兒科支氣管氣喘寒熱證型診斷表》，利用此表將兒科氣喘門診患者區分為寒證、熱證並篩選正常體質健康兒童做為對照組，選擇寒證組及熱證組當中總得分最高的病患及正常體質對照組的周邊血液單核球之mRNA檢體作為寡核?酸晶片的分析樣本。由台大基因微陣列核心實驗室提供晶片相關服務，以雙色螢光標記進行分組樣本的基因比對。結果：比對寒證氣喘組與正常體質對照組樣本，經t-test檢定後，有顯著差異的表達基因共170個，當中包含5個具特殊功能或調控路徑的基因；此5個基因的表達除了與氣喘具有關聯性，亦可能影響氣喘患者的證型表現，使其傾向表現寒證氣喘證型。比對熱證氣喘組與正常體質對照組樣本，經t-test檢定後，有顯著差異的表達基因共266個，當中包含17個具特殊功能或調控路徑的基因，此群基因的表達除了與氣喘具有關聯性，亦可能影響氣喘患者的證型表現，使其傾向表現熱證氣喘證型。比對寒證氣喘組與熱證氣喘組樣本，經t-test檢定後，有顯著差異的表達基因共137個，當中包含8個具特殊功能或調控路徑的基因，由於此二族群的共同條件為氣喘患者，暗示此8個基因的表達與中醫的寒性證型、熱性證型具關聯性。證實寒證氣喘病、熱證氣喘病確實存在不同的基因表現及調控路徑；中醫的寒性證型、熱性證型也確實存在不同的基因表現及調控路徑。 關鍵字：小兒、氣喘、寒證、熱證、證型、基因、寡核?酸晶片; Transcription Analysis in Asthmatic Children with Han Zheng or Re Zheng by Oligonucleotide Microarray Fang-Hui Wei Major professor: Shung-Te Kao Institute of Chinese Medical Science, China Medical University Background: Allergic asthma is a chronic inflammatory disease of lungs. It is also a common disease in pediatrics. Asthma results from a complex interplay between genetic and environmental factors. According to traditional Chinese medicine, allergic asthma can be differentiated between Han Zheng and Re Zheng. Objective: To Analyze the gene expression in Asthmatic children with Han Zheng or Re Zheng using oligonucleotide microarray. Methods: Firstly, we designed a pediatrician examination chart for allergic asthma of Han Zheng or Re Zheng. Based on the chart, we recruited typical patients with Han Zheng or Re Zheng asthma. Healthy children were considered as normal controls. Patients’ mRNAs in their peripheral blood were used for microarray analysis. More than 13,000 genes were compared. Pathway of the differentially expressed genes with the bioinformation software on line was investigated. Result: We found there were 170 differentially expressed genes between Han Zheng asthma group (n = 2) and control group (n = 2), including 5 genes with associated pathway (P < 0.05). There were 226 differentially expressed genes between Re Zheng asthma group (n = 4) and the controls (n = 4), including 17 genes with associated pathway (P < 0.05). There were 137 differentially expressed genes between Han Zheng asthma group (n = 4) and Re Zheng asthma group (n = 4), including 8 genes with associated pathway (P < 0.05). Conclusion: The gene expression of allergic asthma could be correlated with immune and Zheng points. We believe that some genes are associated with Han Zheng or Re Zheng. These differentially expressed genes could be potential molecular targets to treat asthma with traditional Chinese medicine. Key words: pediatrics, allergic asthma, Han Zheng, Re Zheng, microarray
