三七总皂苷对高糖状态下巨噬细胞胆固醇转运基因表达的影响  

Effects of Panax notoginseng saponins on expression of cholesterol transportation gene of macrophages under hyperglycemic condition

作　　者：郑建雷[1] 王利宏[1] 王海清[1] 孙渴望 ZHENG Jian-lei;WANG Li-hong;WANG Hai-qing;SUN Ke-wang(Department of Cardiology,People ＂s Hospital of Hangzhou Medical Colleg;Zhejiang Provincial People＇s Hospital,Hangzhou 310014,China)

机构地区：[1]浙江省人民医院杭州医学院附属人民医院心内科,杭州310014

出　　处：《中国临床药理学杂志》2018年第19期2335-2338,共4页The Chinese Journal of Clinical Pharmacology

基　　金：浙江省中医药优秀青年人才计划基金资助项目(2013ZQ003);浙江省医药卫生科技基金资助项目(2014KYA026);浙江省医学重点学科基金资助项目(2015-ZDCXXK01-2)

摘　　要：目的研究三七总皂苷(PNS)对高糖环境下巨噬细胞吞噬氧化低密度脂蛋白(ox LDL)、胆固醇转运基因Toll样受体4(TLR-4)、清道夫受体B类I型(SR-BI)和ATP结合盒转运蛋白A1(ABCA1)表达的影响。方法将人单核细胞经佛波脂诱导转化成巨噬细胞,将巨噬细胞分为6组:正常对照组(葡萄糖5.5 mmol·L^(-1))、高糖组(葡萄糖15 mmol·L^(-1))、高糖+ox LDL(葡萄糖15mmol·L^(-1)+100 mg·L^(-1))组、阿托伐他汀组(葡萄糖15 mmol·L^(-1)+阿托伐他汀10μmol·L^(-1))与低、高2个剂量PNS组(葡萄糖15 mmol·L^(-1)+PNS 20,60μg·mL^(-1))。以油红染色观察巨噬细胞内脂质含量,以反转录实时定量PCR和免疫印迹法分别测定巨噬细胞胆固醇转运基因TLR-4、SR-BI和ABCA1mRNA与蛋白的表达水平。结果高糖+oxLDL组和正常对照组的巨噬细胞内脂质含量分别为(36±1)%和(17±1)%,组间差异有统计学意义(P<0.01)。高糖+oxLDL组的TLR-4、SR-BI和ABCA1 mRNA相对表达量分别为2.82±0.22,0.40±0.02和0.38±0.06;正常对照组相应mRNA表达量分别为1.00±0.09,1.00±0.01和1.00±0.06,组间比较差异均有统计学意义(均P<0.01)。高糖+oxLDL组的TLR-4、SR-BI和ABCA1蛋白相对灰度值分别为0.96±0.02,0.24±0.02和0.27±0.01;正常对照组分别为0.34±0.01,0.87±0.02和0.82±0.02,组间比较差异均有统计学意义(均P<0.01)。高剂量PNS干预后,巨噬细胞脂质含量降低到(29±1)%,与高糖+oxLDL组相比,差异有统计学意义(P<0.01);高剂量PNS组的TLR-4、SR-BI和ABCA1的mRNA相对表达量分别为1.55±0.34,0.82±0.08和0.84±0.04;而TLR-4、SR-BI和ABCA1蛋白相对灰度值分别为0.55±0.02,0.70±0.01和0.67±0.02,与高糖+oxLDL组相比,差异均有统计学意义(均P<0.01)。结论 PNS具有上调SR-BI和ABCA1、而降低TLR-4表达、减少巨噬细胞对oxLDL摄取的作用。Objective To explore the effects of Panax notoginseng saponins（ PNS） on the oxidized low-density lipoprotein（ ox LDL） uptake,mRNA and protein expression of Toll-like receptor 4（ TLR-4）,Scavenger receptor class B type I（ SR-BI） and ATP-binding cassette transporter A1（ ABCA1） of macrophages cultured under hyperglycemic condition. Methods Human acute monocyte leukaemia cell line（ THP-1） was induced into macrophages with Phorbol ester（ PMA）.Macrophages were then divided into six groups as follows： normal controlgroup（ 5. 5 mmol·L^-1 glucose）,hyperglycemia group（ 15 mmol·L^-1 glucose）,hyperglycemia plus ox LDL group（ 15 mmol·L^-1 glucose ＋ 100 mg·L^-1 ox LDL）,hyperglycemia plus atorvastatin group（ 15 mmol·L^-1 glucose ＋ atorvastatin 10 μmol·L^-1）,hyperglycemia plus PNS groups（ 15 mmol·L^-1 glucose ＋ 20,60 μg · mL^-1 PNS,respectively）.The content of lipid in macrophages was evaluated with oil red O staining. The mRNA levels of TLR-4,SR-BI and ABCA1 were measured by reverse transcription-quantitative polymerase chain reaction,and the protein concentrations of TLR-4,SR-BI and ABCA1 were evaluated by Western blot. Results The lipid contents of hyperglycemia plus ox LDL group and normal control group were（ 36 ± 1） %,（ 17 ± 1） %,the difference was significant（ P〈0. 01）. The relative mRNA levels of TLR-4,SR-BI and ABCA1 in hyperglycemia plus ox LDL group were 2. 82 ± 0. 22,0. 40 ± 0. 02 and 0. 38 ± 0. 06,respectively; which in normal control group were 1. 00 ± 0. 09,1. 00 ± 0. 01 and 1. 00 ± 0. 06,respectively; comparison between the 2 groups,the difference of the three factors were significant（ all P〈0. 01）. The relative protein grey values of TLR-4,SR-BI and ABCA1 in hyperglycemia plus oxLDL group were0. 96 ± 0. 02,0. 24 ± 0. 02 and 0. 27 ± 0. 01,respectively; while in normal control group they were 0. 34 ± 0. 01,0. 87 ± 0. 02 and 0. 82 ± 0. 02,respectively; the difference of the three factors between the 2 groups were signi

关 键 词：三七总皂苷 高血糖 氧化低密度脂蛋白 巨噬细胞 胆固醇转运基因 

分 类 号：R972.6[医药卫生—药品]