炎调方抑制LPS诱导人肺泡上皮细胞炎症因子释放机制研究
|
摘要
目的体外观察炎调方通过NF-κB信号通路调控内毒素(LPS)诱导的人肺泡上皮(A549)细胞中炎症因子的释放。方法采用炎调方药物血清方法,观察其对A549细胞的作用效果及其机制。MTT法观察细胞存活率;双抗体夹心酶联合免疫吸附法(ELISA法)检测细胞中白介素-8(IL-8)和前列腺素E_2(PGE_2)的表达;Western blot法检测环加氧酶2(COX2)和NF-κB的表达;运用NF-κB特异性抑制剂与炎调方联合运用,观察NF-κB信号通路对COX2的调控作用。结果 LPS明显抑制了A549细胞增殖,炎调方药物血清(炎调方药物血清)能缓解LPS的细胞毒性;炎调方药物血清明显降低了LPS诱导的IL-8、PGE_2和COX2释放和表达。炎调方药物血清通过NF-κB信号通路调控COX2表达。结论炎调方通过NF-κB信号通路调控LPS的细胞毒性和炎症因子释放。
Objective:To observe the release of inflammatory cytokines in the LPS induced human alveolar epithelial(A549)cells through NF-κB signaling pathway in vitro.Methods:The effect and mechanism of A549 cells were observed by the method of drug serum with Yantiaofang.Cell survivals were observed by MTT assay.The levels of IL-8 and PGE_2 were detected by ELISA assay.The expressions of COX2 and NF-κB were detected by Western blot analysis.The regulatory effect of NF-κB signaling pathway on COX2 was observed by using the combination of the specific inhibitor of NF-κB and Yantiaofang.Results:LPS significantly inhibited the proliferation of A549 cells.Moreover,the cytotoxicity of LPS can alleviate by the drug serum with Yantiaofang(YTF).YTF significantly reduced the release of IL-8,PGE_2 and COX2 release induced by LPS.The expression of COX2 was regulated by YTF through NF-κB signaling pathway.Conclusion:Yantiaofang regulates LPS induced cytotoxicity and inflammatory cytokine release by NF-κB signaling pathway.
Objective:To observe the release of inflammatory cytokines in the LPS induced human alveolar epithelial(A549)cells through NF-κB signaling pathway in vitro.Methods:The effect and mechanism of A549 cells were observed by the method of drug serum with Yantiaofang.Cell survivals were observed by MTT assay.The levels of IL-8 and PGE_2 were detected by ELISA assay.The expressions of COX2 and NF-κB were detected by Western blot analysis.The regulatory effect of NF-κB signaling pathway on COX2 was observed by using the combination of the specific inhibitor of NF-κB and Yantiaofang.Results:LPS significantly inhibited the proliferation of A549 cells.Moreover,the cytotoxicity of LPS can alleviate by the drug serum with Yantiaofang(YTF).YTF significantly reduced the release of IL-8,PGE_2 and COX2 release induced by LPS.The expression of COX2 was regulated by YTF through NF-κB signaling pathway.Conclusion:Yantiaofang regulates LPS induced cytotoxicity and inflammatory cytokine release by NF-κB signaling pathway.
引文
[1]Galley HF.Oxidative stress and mitochondrial dysfunction in sepsis[J].Br J Anaesth,2011,107(1):57-64.
[2]施荣,熊旭东.NF-κB在脓毒症大鼠肺组织中的活性变化及炎调方对其活性的影响[J].时珍国医国药,2014,25(4):1016-1018.
[3]施荣,熊旭东,李淑芳,等.炎调方对脓毒症大鼠血清炎症因子的调控作用[J].中国中医急症,2012,21(3):397-398.
[4]Rubenfeld GD,Caldwell E,Peabody E,et al.In cidence and outcomes of acute lung injury[J].N Engl J Med,2005,353(16):1685-1693.
[5]Beasley MB.The pathologist's approach to acute lung injury[J].Arch Pathol Lab Med,2010,134(5):719-727.
[6]林栋栋,孙家邦,李非,等.IL-8、IL-10在大鼠急性胰腺炎并发肺损伤中的作用[J].首都医科大学学报,2005,26(2):193-196.
[7]顾言,陈建荣,邵峰,等.清肺汤对急性呼吸窘迫综合征患者呼出气冷凝液和血清中内皮素-1、前列腺素E2的影响[J].实用医学杂志,2014,43(23):3853-3855.
[8]Malek HA,Saleh DM.Cyclooxygenase-2 inhibitor celecoxib in a rat model of hindlimb ischemia reperfu sion[J].Can J Physiol Pharmacol,2009,87(5):353-359.
[9]Schmidt C,Kurt B,Hocherl K,et al.Inhibition of NF-kappa B activity prevents downregulation of alpha1-adrenergic receptors and circulatory failure during CLP-induced sepsis[J].Shock,2009,32(3):239-246.
[10]张云松,朱晓林.脓毒症中医病机及治法探讨[J].上海中医药杂志,2012,46(10):8-9.
[11]施荣,熊旭东.通腑清营法治疗脓毒症热毒内盛证临床观察[J].中国中医急症,2011,20(4):521-522.
[2]施荣,熊旭东.NF-κB在脓毒症大鼠肺组织中的活性变化及炎调方对其活性的影响[J].时珍国医国药,2014,25(4):1016-1018.
[3]施荣,熊旭东,李淑芳,等.炎调方对脓毒症大鼠血清炎症因子的调控作用[J].中国中医急症,2012,21(3):397-398.
[4]Rubenfeld GD,Caldwell E,Peabody E,et al.In cidence and outcomes of acute lung injury[J].N Engl J Med,2005,353(16):1685-1693.
[5]Beasley MB.The pathologist's approach to acute lung injury[J].Arch Pathol Lab Med,2010,134(5):719-727.
[6]林栋栋,孙家邦,李非,等.IL-8、IL-10在大鼠急性胰腺炎并发肺损伤中的作用[J].首都医科大学学报,2005,26(2):193-196.
[7]顾言,陈建荣,邵峰,等.清肺汤对急性呼吸窘迫综合征患者呼出气冷凝液和血清中内皮素-1、前列腺素E2的影响[J].实用医学杂志,2014,43(23):3853-3855.
[8]Malek HA,Saleh DM.Cyclooxygenase-2 inhibitor celecoxib in a rat model of hindlimb ischemia reperfu sion[J].Can J Physiol Pharmacol,2009,87(5):353-359.
[9]Schmidt C,Kurt B,Hocherl K,et al.Inhibition of NF-kappa B activity prevents downregulation of alpha1-adrenergic receptors and circulatory failure during CLP-induced sepsis[J].Shock,2009,32(3):239-246.
[10]张云松,朱晓林.脓毒症中医病机及治法探讨[J].上海中医药杂志,2012,46(10):8-9.
[11]施荣,熊旭东.通腑清营法治疗脓毒症热毒内盛证临床观察[J].中国中医急症,2011,20(4):521-522.