摘要
目的探讨五甲基槲皮素对急性加重慢性阻塞性肺疾病(acute exacerbation of chronic obstructive pulmonary disease,AECOPD)的预防作用及相关机制。方法采用香烟烟雾提取物刺激人支气管上皮BEAS-2B细胞,建立AECOPD细胞模型。采用不同浓度五甲基槲皮素干预AECOPD细胞模型,分为4组:对照组、五甲基槲皮素1μmol/L组、五甲基槲皮素3μmol/L组和五甲基槲皮素10μmol/L组。干预结束后收集细胞蛋白及上清液。采用ELISA实验检测上清液中IL-6、IL-8的浓度;采用蛋白免疫印迹法检测各组样本中磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(proteinkinase B,AKT)、P38蛋白激酶(P38 mitogen-activated protein kinase,P38)、应激活化蛋白激酶(c-Jun N-terminal kinase,JNK)、B-细胞淋巴瘤因子2(B-cell lymphoma-2,BCL-2)、半胱氨酸蛋白酶3(Caspase 3)以及活化后半胱氨酸蛋白酶3(cleaved Caspase 3)的蛋白丰度。结果五甲基槲皮素可剂量依赖性降低IL-6和IL-8的释放,与对照组相比,五甲基槲皮素3μmol/L组和五甲基槲皮素10μmol/L组对IL-6的释放抑制更为明显(P<0.01),五甲基槲皮素10μmol/L组IL-8水平明显降低(P<0.01);同时,五甲基槲皮素10μmol/L组PI3K(P<0.01)、AKT(P<0.01)、P38(P<0.05)、JNK(P<0.05)、Caspase 3(P<0.01)以及cleaved Caspase 3(P<0.001)的蛋白水平降低,并且BCL-2的蛋白表达增加(P<0.01)。结论五甲基槲皮素可通过PI3K/AKT、P38和JNK等信号通路改善AECOPD模型中细胞的炎性反应,同时可通过调节Caspase 3和BCL-2抑制细胞的凋亡,提示五甲基槲皮素具有预防AECOPD发生发展的潜在药理作用。
Objective To investigate the preventive effect of pentamethylquercetin on acute exacerbation of chronic obstructive pulmonary disease(AECOPD)and its related mechanism.Methods Cigarette smoke extract was used to stimulated human bronchial epithelial BEAS-2B cells to establish AECOPD cell model,and divided into 4 groups:control group,pentamethylquercetin 1μmol/L group,pentamethylquercetin 3μmol/L group and pentamethylquercetin 10μmol/L group.Cell protein and supernatant were collected after intervention.The content of IL-6 and IL-8 in supernatant were detected by ELISA.The protein expression levels of phosphatidylinositol 3-kinase(PI3K),proteinkinase B(AKT),P38 mitogen-activated protein kinase(P38),c-Jun N-terminal kinase(JNK),B-cell lymphoma-2(BCL-2),Caspase 3 and cleaved Caspase 3 were detected by Western blot.Results Pentamethylquercetin can reduce the release of IL-6 and IL-8 in a dose-dependent manner.Compared with control group,the release of IL-6 was significantly inhibited in pentamethylquercetin 3μmol/L group and pentamethylquercetin 10μmol/L group(P<0.01),and IL-8 level in pentame-thylquercetin 10μmol/L group decreased(P<0.01).Meanwhile,pentamethylquercetin 10μmol/L group decreased the expression of PI3K(P<0.01),AKT(P<0.01),P38(P<0.05),JNK(P<0.05),Caspase 3(P<0.01)and cleaved Caspase 3(P<0.001),and increased BCL-2 protein level(P<0.01).Conclusion Pentamethylquercetin can improve the inflammatory reaction in AECOPD model via inhibiting PI3K/AKT,P38 and JNK pathway,and can inhibit cell apoptosis by modulating Caspase 3 and BCL-2,which indicated the potential pharmacological effect of pentamethylquercetin on AECOPD.
作者
许夏燕
李智
潘明月
韩雨彤
易刚强
袁劲松
XU Xiayan;LI Zhi;PAN Mingyue;HAN Yutong;YI Gangqiang;Yuan Jingsong(Shantou Medical University,Shantou,Guangdong 515041,China;Shenzhen Luohu District People's Hospital,Shenzhen,Guangdong 518000,China;Hunan University of Chiese Medicine,Changsha,Hunan 410208,China;Shenzhen Hospital of Beijing University,Shenzhen,Guangdong 518000,China)
出处
《湖南中医药大学学报》
CAS
2022年第8期1283-1288,共6页
Journal of Hunan University of Chinese Medicine
基金
长沙市自然科学基金项目(kq2007040)
深圳市罗湖区软科学研究计划项目(LX20201101)。
