TLR4表达水平与COPD大鼠肺动脉平滑肌细胞合成分泌功能的关系

发布时间：2018-05-21 15:56

本文选题：慢性阻塞性肺疾病 + 肺动脉平滑肌细胞　；参考：《安徽医科大学学报》2017年09期

【摘要】：目的探讨Toll样受体4(TLR4)表达水平与慢性阻塞性肺疾病(COPD)大鼠肺动脉平滑肌细胞(PASMCs)合成分泌功能的关系。方法建立大鼠COPD模型,HE染色判定模型建立,免疫组织化学染色观察对照组与COPD模型组肺组织动脉平滑肌层TLR4的表达。分离培养鉴定原代PASMCs,脂多糖(LPS)、TLR4的特异性抑制剂TAK-242干预细胞,实验分组:空白组、LPS组、TAK-242组、LPS+TAK-242组,Western blot检测各组PASMCs中TLR4的表达,ELISA法检测各组细胞培养上清液中Th1细胞因子干扰素(IFN)-γ、Th2细胞因子白介素(IL)-6及血小板衍化生长因子(PDGF)的浓度;并将TLR4的表达水平与上清液中炎性因子的分泌水平进行相关性分析。结果 COPD模型组大鼠肺组织动脉平滑肌层TLR4的表达明显高于正常大鼠;与空白组比较,LPS组PASMCs中TLR4的表达明显升高(P0.05);LPS组PASMCs中合成分泌IFN-γ、IL-6、PDGF的水平较对照组明显升高,TAK-242阻断TLR4,PASMCs中TLR4的表达明显降低(P0.05);细胞培养上清液中IFN-γ、IL-6、PDGF的表达水平较空白组明显降低(P0.05);TLR4表达水平与细胞培养上清液IFN-γ、IL-6及PDGF的浓度呈显著正相关性(r=0.95、0.87、0.83,P0.05)。结论TLR4可能参与调控PASMCs的合成分泌功能。
[Abstract]:Objective to investigate the relationship between the expression of Toll like receptor 4 (TLR4) and the synthesis and secretion of pulmonary artery smooth muscle cells (PASMCs) in rats with chronic obstructive pulmonary disease (COPD). Methods the rat COPD model was established by HE staining and the expression of TLR4 in the smooth muscle layer of pulmonary artery in the control group and the COPD model group was observed by immunohistochemical staining. Primary PASMCs, a specific inhibitor of lipopolysaccharide TLR4, TAK-242, were isolated and identified. The expression of TLR4 in PASMCs was detected by Western blot. The concentration of Th1 cytokine IFN- 纬 -Th2 cytokine Interleukin-6 and platelet-derived growth factor (PDGF) in supernatant of each group was detected by Elisa. The correlation between the expression of TLR4 and the secretion of inflammatory factors in supernatant was analyzed. Results the expression of TLR4 in the smooth muscle layer of pulmonary artery in COPD group was significantly higher than that in normal rats. Compared with the control group, the expression of TLR4 in PASMCs in the LPS group was significantly higher than that in the control group, and the expression of IFN- 纬 -IL-6 mRNA in PASMCs was significantly higher than that in the control group. The expression of IFN- 纬 -IL-6 mRNA in the supernatant of cell culture was significantly lower than that in the control group, and the expression level of IFN- 纬 -IL-6 mRNA in the supernatant of cell culture was significantly lower than that in the control group. There was a significant positive correlation between the expression of TLR4 and the concentration of IL-6 and PDGF in the supernatant of cell culture. Conclusion TLR4 may be involved in regulating the synthesis and secretion of PASMCs.
【作者单位】：桂林医学院附属医院呼吸内科;
【基金】：国家自然科学基金(编号:81360010) 广西医疗卫生适宜技术开发与推广应用项目(编号:S201603)
【分类号】：R563.9

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