ADAM33在IL-4刺激的人气道平滑肌细胞中对NF-κB和TGF-β1表达的影响

发布时间：2018-02-20 15:08

本文关键词： ADAM IL- NF-κB TGF-β 气道重塑　出处：《基础医学与临床》2014年10期 　论文类型：期刊论文

【摘要】：目的探讨解整合素-基质金属蛋白酶33(ADAM33)在白介素-4(IL-4)刺激的人气道平滑肌细胞(HASMCs)中对NF-κB和TGF-β1表达的影响,为进一步完善哮喘的发病机制提供理论基础。方法以50μg/L的IL-4刺激HASMCs,以未刺激细胞组为对照,24 h后实时荧光定量PCR法和Western blot法检测ADAM33的表达;设计并合成ADAM33-siRNA,IL-4刺激24 h后瞬时转染ADAM33-siRNA,实时荧光定量PCR法和Western blot法检测抑制率;并且检测TGF-β1、NF-κB mRNA和蛋白的表达情况。结果 50μg/L的IL-4较对照组明显促进了HASMCs中ADAM33的表达(P0.05),在mRNA水平表达量增加约4.36倍,蛋白水平增加约3.3倍;ADAM33-siRNA-575明显抑制了ADAM33在HASMCs中的表达;干扰组、阴性对照组和空白对照组TGF-β1 mRNA的相对表达量分别为0.602±0.024、1.01±0.176和1.239±0.171,NF-κB mRNA的表达量分别为0.54±0.08、1.014±0.21和1.049±0.378;TGF-β1蛋白的表达量分别为0.227±0.016、0.7±0.048和0.715±0.025,NF-κB蛋白的表达量分别为0.335±0.048、0.922±0.04和0.943±0.046。干扰组TGF-β1、NF-κB不论是在mRNA水平,还是在蛋白水平,均明显低于阴性对照组和空白对照组(P0.05)。结论 ADAM33可能是IL-4调节人气道平滑肌细胞NF-κB/TGF-β1信号通路中的一个关键信号分子,有望成为防治哮喘的重要靶点。
[Abstract]:Objective to investigate the effect of integrin-matrix metalloproteinase-33 (ADAM33) on the expression of NF- 魏 B and TGF- 尾 1 in human airway smooth muscle cells (HASMCs) stimulated by interleukin-4 (IL-4). Methods HASMCs were stimulated with 50 渭 g / L IL-4 and the expression of ADAM33 was detected by real-time fluorescence quantitative PCR and Western blot after 24 h of control group. ADAM33-siRNA-IL-4 was designed and synthesized for 24 h, then transfected into ADAM33-siRNAs for 24 h. The inhibition rate was detected by real-time fluorescence quantitative PCR assay and Western blot assay. Results 50 渭 g / L IL-4 significantly promoted the expression of ADAM33 in HASMCs, increased the expression of ADAM33 by 4.36 times at mRNA level, and increased the protein level by about 3.3 times. ADAM33-siRNA-575 significantly inhibited the expression of ADAM33 in HASMCs. The relative expression levels of TGF- 尾 1 mRNA in negative control group and blank control group were 0.602 卤0.024 1.01 卤0.176 and 1.239 卤0.171 respectively. The expression of TGF- 尾 1 mRNA was 0.54 卤0.081.014 卤0.21 and 1.049 卤0.378TGF- 尾 1 protein in negative control group and blank control group, respectively 0.227 卤0.0160.7 卤0.048 and 0.715 卤0.025 NF-kappa B protein expression was 0.335 卤0.080.922 卤0.04 and 0.943 卤0.0460.The expression of TGF- 尾 _ 1NF- 魏 B in the interfering group was 0.335 卤0.080.922 卤0.04 and 0.943 卤0.0466.The expression levels of TGF- 尾 _ 1NF- 魏 B were 0.227 卤0.0160.7 卤0.048 and 0.715 卤0.025 respectively. Conclusion ADAM33 may be a key signal molecule in the regulation of NF- 魏 B / TGF- 尾 1 signaling pathway by IL-4, and may be an important target in the prevention and treatment of asthma.
【作者单位】：重庆市涪陵中心医院呼吸内科;
【基金】：重庆市卫生局项目(2007-2-222)
【分类号】：R562.25

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