原儿茶酸抑制结核分枝杆菌Rv1654诱导肺泡巨噬细胞凋亡的机制研究  

Protocatechuic acid inhibits apoptosis of Alveolar macrophages induced by Mycobacterium tuberculosis Rv1654

作　　者：林卫佳 张亚平 李峰 项保利[1] 张志华[1] 张秀珑[1] LIN Wei-jia;ZHANG Ya-ping;LI Feng;XIANG Bao-li;ZHANG Zhi-hua;ZHANG Xiu-long(Department of Respiratory and Critical Care Medicine,The First Afiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province,China)

机构地区：[1]河北北方学院附属第一医院呼吸与危重症医学科,河北张家口075000

出　　处：《中国临床药理学杂志》2022年第15期1747-1751,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河北省医学科学研究重点课题计划基金资助项目(20180869)。

摘　　要：目的 探讨原儿茶酸对结核分枝杆菌Rv1654诱导肺泡巨噬细胞凋亡影响和机制。方法 肺泡巨噬细胞NR8383分成对照组、模型组(结核分枝杆菌Rv1654诱导)、实验低剂量组(结核分枝杆菌Rv1654诱导,20μmol·L^(-1)原儿茶酸处理)、实验中剂量组(结核分枝杆菌Rv1654诱导,40μmol·L^(-1)原儿茶酸处理)、实验高剂量组(结核分枝杆菌Rv1654诱导,80μmol·L^(-1)原儿茶酸处理)、实验高剂量+Vector组(结核分枝杆菌Rv1654诱导,转染阴性对照载体,80μmol·L^(-1)原儿茶酸处理)、实验高剂量+TLR2组(结核分枝杆菌Rv1654诱导,转染TLR2过表达载体,80μmol·L^(-1)原儿茶酸处理)。用蛋白质印迹(Western blot)法检测Toll样受体2(TLR2)蛋白的表达水平,用流式细胞术检测细胞凋亡情况,用JC-1法检测线粒体膜电位。结果 对照组、模型组、实验低剂量组、实验中剂量组、实验高剂量组、实验高剂量+Vector组、实验高剂量+TLR2组肺泡巨噬细胞中TLR2蛋白相对表达量分别为0.23±0.04,0.66±0.09,0.53±0.02,0.41±0.03,0.29±0.02,0.31±0.04和0.48±0.05,细胞凋亡率分别为(4.17±0.29)%,(14.41±0.37)%,(10.51±1.02)%,(8.40±0.59)%,(6.77±0.58)%,(6.46±0.32)%和(9.78±0.96)%,线粒体膜电位分别为(17.30±1.63)%,(6.47±0.49)%,(8.30±0.85)%,(10.31±1.37)%,(14.27±1.68)%,(14.00±1.06)%和(8.66±0.60)%,模型组的上述指标与对照组比较,差异均有统计学意义(均P<0.05);实验低、中、高剂量组的上述指标与模型组比较,差异均有统计学意义(均P<0.05);实验低、中剂量组的上述指标比较,差异均有统计学意义(均P<0.05);实验中剂量组与实验高剂量组的上述指标比较,差异均有统计学意义(均P<0.05);实验高剂量+Vector组的上述指标与实验高剂量+TLR2组比较,差异均有统计学意义(均P<0.05)。结论 原儿茶酸通过下调TLR2抑制线粒体途径减少结核分枝杆菌Rv1654诱导肺泡巨噬细胞凋亡。Objective To investigate the effect and mechanism of protocatechuic acid on the apoptosis of alveolar macrophages induced by Mycobacterium tuberculosis Rv1654.Methods Alveolar macrophages NR8383 were divided into control group,model group (induction of Mycobacterium tuberculosis Rv1654),experimental low-dose group(induction of Mycobacterium tuberculosis Rv1654,20μmol·L^(-1)protocatechuic acid treatment),and experimental medium-dose group(induction of Mycobacterium tuberculosis Rv1654,40μmol·L^(-1) protocatechuic acid treatment),experimental high-dose group (induction of Mycobacterium tuberculosis Rv1654,80μmol·L^(-1) protocatechuic acid treatment),experimental high-dose+Vector group (induction of Mycobacterium tuberculosis Rv1654,transfection with negative control vector,80μmol·L^(-1) protocatechuic acid treatment),experimental high-dose+TLR2 group (induction of mycobacterium tuberculosis Rv1654,transfection of TLR2overexpression vector,80μmol·L^(-1) protocatechuic acid treatment),Western blot method was used to detect Tolllike receptor 2 (TLR2) protein expression,flow cytometry was used to detect apoptosis,JC-1 method was performed to detect mitochondrial membrane potential.Results The expression of TLR2 protein in alveolar macrophages in control group,model group,experimental low-dose group,experimental medium-dose group,experimental high-dose group,experimental high-dose+Vector group,experimental high-dose+TLR2 group were 0.23±0.04,0.66±0.09,0.53±0.02,0.41±0.03,0.29±0.02,0.31±0.04 and 0.48±0.05,the apoptosis rate were (4.17±0.29)%,(14.41±0.37)%,(10.51±1.02)%,(8.40±0.59)%,(6.77±0.58)%,(6.46±0.32)%and(9.78±0.96)%,mitochondrial membrane potential were (17.30±1.63)%,(6.47±0.49)%,(8.30±0.85)%,(10.31±1.37)%,(14.27±1.68)%,(14.00±1.06)%and (8.66±0.60)%.Compared model group with control group,the differences of the above indicators were all statistically significant (all P<0.05);compared experimental low-dose group,experimental medium-dose group,experimental high-dose group with model gro

关 键 词：原儿茶酸 结核分枝杆菌 线粒体途径 巨噬细胞 凋亡 

分 类 号：R28[医药卫生—中药学]