RNA 干擾靶向抑制酪氨酸蛋白激酶對哮喘小鼠樹突狀細胞功能的影響_《中國呼吸與危重監護雜志》

作者：

鄭永華 ¹ , 陸晶晶 ² , 郭忠良 ² , 任濤 ² ,  梁永杰 ²

1 東陽市人民醫院呼吸內科(浙江東陽 322100)2 同濟大學附屬東方醫院呼吸內科(上海 200120);

關鍵詞：

哮喘小干擾RNA 樹突狀細胞酪氨酸蛋白激酶細胞因子

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目的通過特異性微小干擾RNA( siRNA) 靶向抑制小鼠骨髓來源樹突狀細胞( DC) 的酪氨酸蛋白激酶( Syk) 的基因表達, 阻斷樹突狀細胞的成熟, 從而阻斷Syk 所介導的細胞內及細胞間的信號傳導通路, 研究Syk 與DC 之間的關系。方法化學法合成21 ～23 bp 的小鼠樹突狀細胞Syk 基因siRNA 片段A、B、C、D, 用Lipofectamine 2000 作為轉染試劑, 將siRNA 片段轉染哮喘小鼠骨髓來源DC, 作用48 h 后再與正常小鼠脾臟來源T淋巴細胞混合反應48 h, 之后用ELISA 法檢測T 淋巴細胞分泌細胞因子( IL-4、IL-13、IL-2 和INF-γ) 的水平。用Western Blot 檢測DC 表達Syk 蛋白的量, 判斷Syk 基因是否被沉默, 剔除無效片段。結果 siRNA 片段A、C、D 干擾組小鼠DC 細胞Syk 蛋白表達減少; B組沒有明顯變化, 為無效片段。siRNA 干擾組IL-4、IL-13 的水平較未干擾組明顯下降( P 均 lt;0. 05) , IL-2 和INF-γ水平無顯著差異( P gt;0. 05) 。結論特異性SiRNA 能夠有效抑制Syk 基因表達, 可阻遏DC 的抗原遞呈作用以及活化、分化T 淋巴細胞的功能

引用本文： 鄭永華,陸晶晶,郭忠良,任濤,梁永杰. RNA 干擾靶向抑制酪氨酸蛋白激酶對哮喘小鼠樹突狀細胞功能的影響. 中國呼吸與危重監護雜志, 2009, 09(5): 487-491. doi: 復制

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10.	, No.
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13.	Cohn L, Elias JA, Chupp GL. Asthma: mechanisms of disease persistence and progression. Annu Rev Immunol, 2004 , 22: 789 - 815.
14.	Nadler MJ, Matthews SA, Turner H, et al. Signal transduction by the high-affinity immunoglobulin E receptor FcqRI: coupling form to function. Adv Immunol, 2000, 76: 325-355.
15.	Nel AE. T-cell activation through the antigen receptor. Part 1 : Signaling components, signaling pathways, and signal integration at the T-cell antigen receptor synapse. J Allergy Clin Immunol, 2002 , 109: 758-770.
16.	Samelson LE. Signal transduction mediated by the T cell antigen receptor: the role of adapter proteins. Annu Rev Immunol, 2002, 20: 371-394.
17.	Gauld SB, Dal Porto JM, Cambier JC. B cell antigen receptor signaling: roles in cell development and disease. Science, 2002 , 296: 1641-1642.
18.	Hong JJ, Yankee TM, Harrison ML, et al. Regulation of signaling in B cells through the phosphorylation of Syk on linker region tyrosines. J Biol Chem, 2002, 277: 31703-31714.
19.	Matsubara S, Koya T, Takeda K, et al. Syk activation in dendritic cells is essential for airway hyperresponsiveness and inflammation. Am J Respir Cell Mol Biol, 2006, 34: 426-433.
20.	emann UA, Ray P, FlaveII RA. Pulmonary overexpression of IL-9 induces Th2 cytokine expression, leading to immune pathology. J Clin Invest, 2002, 109: 29-39.
21.	毛光宇, 楊炯, 陳宏斌, 等. 過敏性哮喘患者樹突狀細胞對原始T 細胞活化的影響. 中國呼吸與危重監護雜志, 2005, 4: 115-118.
22.	Ma B, Zhu Z, Homer RJ, et al. The C10 /CCL6 chemokine and CCR1 play critical roles in the pathogenesis of IL-13-induced inflammation and remodeling. J Immunol, 2004, 172: 1872-1881.
23.	Zhu Z, Ma B, Zheng T, et al. IL-13-induced chemokine responses in the lung: Role of CCR2 in the pathogenesis of IL-13-induced inflammation and remodeling. J Immunol, 2002, 168: 2953-2962.
24.	Prechtel AT, Turza NM, Theodoridis AA, et al. Small interfering RNA ( siRNA ) delivery into monocyte-derived dendritic cells by electroporation. J Immunol Methods, 2006, 311: 139-152.

1. Elias JA, Zhu Z, Chupp G, et al. Airway remodeling in asthma. J Clin Invest, 1999, 104: 1001-1006.
2. Elias JA, Lee CG, Tao Zet al. New insights into the pathogenesis of ·490· 中國呼吸與危重監護雜志2009 年9 月第8 卷第5 期Chin J Respir Crit Care Med, September 2009 , Vol.
3. 馬艷良, 何權瀛. 從哮喘的流行病學調查看我國哮喘診斷不足問題. 中國呼吸與危重監護雜志, 2004, 3: 10-14.
4. 劉春濤, 馮玉麟, 梁宗安, 等. 支氣管哮喘防治現狀與對策的衛生經濟學分析. 中國呼吸與危重監護雜志, 2002, 1: 148-150.
5. 鐘南山, 主編. 支氣管哮喘與臨床. 北京: 人民衛生出版社, 2006, 450-451.
7. Guermonprez P, Valladeau J, Zitvogel L, et al. Antigen presentation and T cell stimulation by dendritic cells. Annu Rev Immunol, 2002 , 20: 621-667.
8. Hannon GJ. RNA interference. Nature, 2002, 418: 244-251.
9. Plasterk RHA. RNA Silencing: The Genome’s Immune System. Science, 2002, 296: 1263-1265.
10. , No.
11. Wong WS, Leong KP. Tyrosine kinase inhibitors: a new approach for asthma. Biochim Biophys Acta, 2004, 1697: 53-69.
12. 程曉明, 王長征, 李淑平, 等. 哮喘致敏小鼠模型的建立及脾臟 CD4 + T 細胞的分離. 第三軍醫大學學報, 2001, 23: 369-370.
13. Cohn L, Elias JA, Chupp GL. Asthma: mechanisms of disease persistence and progression. Annu Rev Immunol, 2004 , 22: 789 - 815.
14. Nadler MJ, Matthews SA, Turner H, et al. Signal transduction by the high-affinity immunoglobulin E receptor FcqRI: coupling form to function. Adv Immunol, 2000, 76: 325-355.
15. Nel AE. T-cell activation through the antigen receptor. Part 1 : Signaling components, signaling pathways, and signal integration at the T-cell antigen receptor synapse. J Allergy Clin Immunol, 2002 , 109: 758-770.
16. Samelson LE. Signal transduction mediated by the T cell antigen receptor: the role of adapter proteins. Annu Rev Immunol, 2002, 20: 371-394.
17. Gauld SB, Dal Porto JM, Cambier JC. B cell antigen receptor signaling: roles in cell development and disease. Science, 2002 , 296: 1641-1642.
18. Hong JJ, Yankee TM, Harrison ML, et al. Regulation of signaling in B cells through the phosphorylation of Syk on linker region tyrosines. J Biol Chem, 2002, 277: 31703-31714.
19. Matsubara S, Koya T, Takeda K, et al. Syk activation in dendritic cells is essential for airway hyperresponsiveness and inflammation. Am J Respir Cell Mol Biol, 2006, 34: 426-433.
20. emann UA, Ray P, FlaveII RA. Pulmonary overexpression of IL-9 induces Th2 cytokine expression, leading to immune pathology. J Clin Invest, 2002, 109: 29-39.
21. 毛光宇, 楊炯, 陳宏斌, 等. 過敏性哮喘患者樹突狀細胞對原始T 細胞活化的影響. 中國呼吸與危重監護雜志, 2005, 4: 115-118.
22. Ma B, Zhu Z, Homer RJ, et al. The C10 /CCL6 chemokine and CCR1 play critical roles in the pathogenesis of IL-13-induced inflammation and remodeling. J Immunol, 2004, 172: 1872-1881.
23. Zhu Z, Ma B, Zheng T, et al. IL-13-induced chemokine responses in the lung: Role of CCR2 in the pathogenesis of IL-13-induced inflammation and remodeling. J Immunol, 2002, 168: 2953-2962.
24. Prechtel AT, Turza NM, Theodoridis AA, et al. Small interfering RNA ( siRNA ) delivery into monocyte-derived dendritic cells by electroporation. J Immunol Methods, 2006, 311: 139-152.

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RNA 干擾靶向抑制酪氨酸蛋白激酶對哮喘小鼠樹突狀細胞功能的影響

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RNA 干擾靶向抑制酪氨酸蛋白激酶對哮喘小鼠樹突狀細胞功能的影響

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