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ADAM33在IL-4刺激的人氣道平滑肌細胞中對NF-κB和TGF-β1表達的影響

發(fā)布時間:2018-02-20 15:08

  本文關(guān)鍵詞: ADAM IL- NF-κB TGF-β 氣道重塑 出處:《基礎(chǔ)醫(yī)學與臨床》2014年10期  論文類型:期刊論文


【摘要】:目的探討解整合素-基質(zhì)金屬蛋白酶33(ADAM33)在白介素-4(IL-4)刺激的人氣道平滑肌細胞(HASMCs)中對NF-κB和TGF-β1表達的影響,為進一步完善哮喘的發(fā)病機制提供理論基礎(chǔ)。方法以50μg/L的IL-4刺激HASMCs,以未刺激細胞組為對照,24 h后實時熒光定量PCR法和Western blot法檢測ADAM33的表達;設(shè)計并合成ADAM33-siRNA,IL-4刺激24 h后瞬時轉(zhuǎn)染ADAM33-siRNA,實時熒光定量PCR法和Western blot法檢測抑制率;并且檢測TGF-β1、NF-κB mRNA和蛋白的表達情況。結(jié)果 50μg/L的IL-4較對照組明顯促進了HASMCs中ADAM33的表達(P0.05),在mRNA水平表達量增加約4.36倍,蛋白水平增加約3.3倍;ADAM33-siRNA-575明顯抑制了ADAM33在HASMCs中的表達;干擾組、陰性對照組和空白對照組TGF-β1 mRNA的相對表達量分別為0.602±0.024、1.01±0.176和1.239±0.171,NF-κB mRNA的表達量分別為0.54±0.08、1.014±0.21和1.049±0.378;TGF-β1蛋白的表達量分別為0.227±0.016、0.7±0.048和0.715±0.025,NF-κB蛋白的表達量分別為0.335±0.048、0.922±0.04和0.943±0.046。干擾組TGF-β1、NF-κB不論是在mRNA水平,還是在蛋白水平,均明顯低于陰性對照組和空白對照組(P0.05)。結(jié)論 ADAM33可能是IL-4調(diào)節(jié)人氣道平滑肌細胞NF-κB/TGF-β1信號通路中的一個關(guān)鍵信號分子,有望成為防治哮喘的重要靶點。
[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 鹵0.024 1.01 鹵0.176 and 1.239 鹵0.171 respectively. The expression of TGF- 尾 1 mRNA was 0.54 鹵0.081.014 鹵0.21 and 1.049 鹵0.378TGF- 尾 1 protein in negative control group and blank control group, respectively 0.227 鹵0.0160.7 鹵0.048 and 0.715 鹵0.025 NF-kappa B protein expression was 0.335 鹵0.080.922 鹵0.04 and 0.943 鹵0.0460.The expression of TGF- 尾 _ 1NF- 魏 B in the interfering group was 0.335 鹵0.080.922 鹵0.04 and 0.943 鹵0.0466.The expression levels of TGF- 尾 _ 1NF- 魏 B were 0.227 鹵0.0160.7 鹵0.048 and 0.715 鹵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.
【作者單位】: 重慶市涪陵中心醫(yī)院呼吸內(nèi)科;
【基金】:重慶市衛(wèi)生局項目(2007-2-222)
【分類號】:R562.25

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