丹参素改善脂多糖诱导的支气管上皮细胞损伤研究  

Study on Danshensu improving lipopolysaccharide induced bronchial epithelial cell injury

作　　者：堵钧伟 徐则兰 徐钦星 应可净[1] DU Jun-wei;XU Ze-lan;XU Qin-xing;YING Ke-jing(Department of Respiratory Medicine,Run Run Shaw Hospital,School of Medicine,Zhejiang University,Hangzhou 311400,Zhejiang Province,China;Department of Respiratory Medicine,The First People's Hospital of Fuyang District,Hangzhou 311400,Zhejiang Province,China)

机构地区：[1]浙江大学医学院附属邵逸夫医院呼吸科,浙江杭州311400 [2]杭州市富阳区第一人民医院呼吸科,浙江杭州311400

出　　处：《中国临床药理学杂志》2022年第12期1320-1324,共5页The Chinese Journal of Clinical Pharmacology

基　　金：浙江省基础公益研究计划基金资助项目(LGF21H160017)。

摘　　要：目的探讨丹参素(DSS)对脂多糖(LPS)诱导的支气管上皮16HBE细胞损伤的影响及其机制。方法采用噻唑蓝(MTT)法检测DSS对16HBE细胞活力的影响以筛选无毒性的DSS作用浓度。用LPS诱导构建支气管上皮细胞损伤模型。将细胞分为空白组、模型组和低、中、高剂量实验组。空白组细胞正常培养,模型组以25μg·mL^(-1) LPS诱导细胞24 h,低、中、高剂量实验组分别向LPS诱导的细胞中加入20,40,80μmol·L^(-1) DSS处理24 h。以MTT法检测细胞活力;以流式细胞术检测细胞凋亡情况;以酶联免疫吸附(ELISA)法检测细胞炎症因子分泌和氧化应激指标含量;以二氯二氢荧光素-乙酰乙酸酯(DCFH-DA)法检测细胞内活性氧(ROS)水平;以蛋白质印迹法(Western blot)检测细胞沉默信息调节因子1(SIRT1)、NOD样受体蛋白3(NLRP3)蛋白的表达水平。结果空白组、模型组和低、中、高剂量实验组细胞存活率分别为(96.11±5.14)%,(50.23±4.30)%,(51.12±4.08)%,(60.57±4.56)%和(82.03±5.02)%;这5组的凋亡率分别为(4.36±1.03)%,(29.18±3.01)%,(27.91±3.14)%,(21.47±2.27)%和(15.15±2.03)%;这5组的ROS相对荧光强度分别为1.00±0.00,4.16±0.21,4.01±0.19,3.74±0.15和2.64±0.11;这5组的SIRT1蛋白表达水平分别为0.83±0.06,0.23±0.03,0.25±0.03,0.36±0.03和0.51±0.04;这5组的NLRP3蛋白表达水平分别为0.18±0.02,0.59±0.03,0.57±0.03,0.48±0.03和0.29±0.03。上述指标,模型组与空白组比较,中、高剂量实验组与模型组比较,差异均有统计学意义(均P<0.05)。结论DSS可改善LPS诱导的16HBE细胞损伤,其作用机制可能与减轻SIRT1/NLRP3通路介导的氧化应激和炎症有关。Objective To investigate the effect of Danshensu(DSS)on lipopolysaccharide(LPS)-induced injury of bronchial epithelial 16HBE cells and its mechanism.Methods The effects of DSS on the viability of 16HBE cells were detected by tetramethylazolazol blue(MTT)method,so as to screen the non-toxic concentration of DSS.The injury model of bronchial epithelial cells was induced by LPS.The cells were divided into blank group,model group and experimental-L,-M,-H.The cells in the blank group were cultured normally;the cells in the model group were cultured with 25μg·mL^(-1) LPS induced cells for 24 h;LPS-induced cells were added 20,40 and 80μmol·L^(-1) DSS for 24 h in experimental-L,M,-H groups,respectively.Cell viability was detected by MTT assay;cell apoptosis was detected by flow cytometry;the secretion of inflammatory factors and oxidative stress index were detected by enzyme-linked immunosorbent assay(ELISA);intracellular reactive oxygen species(ROS)were detected by dCFH-DA method;the expression levels of silence information regulator 1(SIRT1)and nod-like receptor protein-3(NLRP3)protein were detected by Western blot.Results The cell survival rates of blank group,model group and experimental-L,M,-H groups were(96.11±5.14)%,(50.23±4.30)%,(51.12±4.08)%,(60.57±4.56)%and(82.03±5.02)%respectively;the apoptosis rates of these 5 groups were(4.36±1.03)%,(29.18±3.01)%,(27.91±3.14)%,(21.47±2.27)%and(15.15±2.03)%respectively;the ROS relative fluorescence intensity of these 5 groups were 1.00±0.00,4.16±0.21,4.01±0.19,3.74±0.15 and 2.64±0.11 respectively;SIRT1 protein expression levels in these 5 groups were 0.83±0.06,0.23±0.03,0.25±0.03,0.36±0.03 and 0.51±0.04 respectively;the NLRP3 protein expression levels of these 5 groups were 0.18±0.02,0.59±0.03,0.57±0.03,0.48±0.03 and 0.29±0.03 respectively.There were statistically significant differences in the above indexes between model group and blank group,and between experimental-M group,experimental-H group and model group(all P<0.05).Conclusions DSS can imp

关 键 词：支气管上皮细胞 脂多糖 丹参素 炎症 氧化应激 

分 类 号：R97[医药卫生—药品]