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目的研究C1q/肿瘤坏死因子相关蛋白9(CTRP9)对哮喘幼鼠气道平滑肌细胞(airway smooth muscle cells,ASMCs)增殖和气道炎症的作用。方法 ELISA检测正常、哮喘儿童和小鼠血清中CTRP9含量;原代培养哮喘小鼠ASMCs,经pcDNA3.1-CTRP9转染后,MTT检测ASMCs增殖,ELISA测定TNF-α和IL-6含量,Western印记检测TLR4和NF-κB p65蛋白表达以及NF-κB p65磷酸化水平;pcDNA3.1-CTRP9转染哮喘小鼠,HE染色观察肺组织炎性细胞的浸润程度;收集小鼠支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF),光镜下检测嗜酸性粒细胞、巨噬细胞和中性粒细胞数目;检测小鼠肺部TNF-α、IL-6、TLR4和NF-κB的表达。结果哮喘儿童和小鼠血清中CTRP9含量低于正常组;CTRP9能抑制ASMCs增殖、炎症因子分泌和TLR4/NF-κB通路活化;CTRP9也能抑制哮喘小鼠肺部炎性细胞浸润、各炎性反应细胞数目、炎症因子分泌和TLR4/NF-κB通路活化。结论 CTRP9能抑制幼年哮喘小鼠ASMCs增殖和炎性反应。
Objective To investigate the effects of C1q / tumor necrosis factor-related protein 9 (CTRP9) on proliferation and airway inflammation in airway smooth muscle cells (ASMCs) of young rats with asthma. Methods The serum levels of CTRP9 in normal asthmatic children and mice were detected by ELISA. Primary cultured ASMCs of asthmatic mice were transfected with pcDNA3.1-CTRP9. The proliferation of ASMCs was detected by MTT assay. The levels of TNF-α and IL-6 were detected by ELISA. The expression of TLR4 and NF-κB p65 protein and the phosphorylation of NF-κB p65 were detected by Western blotting. The asthmatic mice were transfected with pcDNA3.1-CTRP9 and the infiltration of inflammatory cells was observed by HE staining. The bronchoalveolar lavage fluid (BALF). The numbers of eosinophils, macrophages and neutrophils were detected by light microscope. The expressions of TNF-α, IL-6, TLR4 and NF-κB in the lung were detected by light microscopy. Results CTRP9 levels in serum of asthmatic children and mice were lower than those in normal mice. CTRP9 could inhibit the proliferation of ASMCs, the secretion of inflammatory cytokines and the activation of TLR4 / NF-κB pathway. CTRP9 also inhibited the infiltration of inflammatory cells and inflammatory cytokines Reactive cell number, inflammatory factor secretion and TLR4 / NF-κB pathway activation. Conclusion CTRP9 can inhibit the proliferation and inflammatory response of ASMCs in young asthmatic mice.