摘要
目的以人单核细胞株THP-1细胞为基础,观察养心氏对THP-1源性巨噬细胞极化和活化的影响,为养心氏抗动脉粥样硬化机制提供理论依据。方法分别以不同浓度(10mg/L,25mg/L和50mg/L)的氧化型低密度脂蛋白(ox-LDL)刺激THP-1源性巨噬细胞24h,以空白为对照组,用ELISA法测定上清液中的单核细胞趋化蛋白-1(MCP-1)和巨噬细胞移动抑制因子(MIF)的浓度,用流式细胞术检测巨噬细胞表面CD16及CD68的表达。用不同浓度的养心氏(10mg/L,50mg/L和100mg/L)预处理THP-1源性巨噬细胞12h,再用50mg/L的ox-LDL刺激THP-1源性巨噬细胞24h,检测上清液中MIF、MCP-1的浓度以及CD16及CD68的表达情况。结果 ox-LDL能以剂量依赖的方式诱导THP-1源性巨噬细胞分泌MCP-1和MIF,其中50mg/L组表达最高,MCP-1(29.30±1.48)pg/mL,对照组为(5.71±1.94)pg/mL;MIF(18.67±0.15)ng/mL,对照组为(4.61±0.40)ng/mL(P<0.01)。ox-LDL能改变巨噬细胞表面抗原CD16、CD68的表达,50mg/L组能显著下调CD16、CD68的表达(CD16:26.9vs对照组29.8;CD68:22.3vs对照组25.2)。养心氏能够以剂量依赖的方式抑制ox-LDL所致的THP-1源性巨噬细胞分泌MCP-1和MIF,随着养心氏刺激浓度的增加,MCP-1和MIF的分泌减少。结论养心氏可能通过抑制巨噬细胞炎性活化,抑制巨噬细胞向促炎表型转化进而抑制动脉粥样硬化的形成和发展。
Objective To explore the anti atherosclerotic mechanism of Yangxinshi tablets（YXST）,we stimulated THP 1 derived macrophages with YXST and observed its effects on macrophage polarity switch and activation.Methods Human monocytic cel line THP 1 were differentiated into macrophages by stimulated with 160 nmol/L phorbol 12 myristate 13 acetate （PMA）for 24 h.Then THP 1 derived macrophages were stimulated by different concentrations of oxidized low density lipoprotein （ox LDL）for 24 h, which were 10mg/L,25mg/L and 50mg/L,and blank was used as control.The secretion of monocyte chemotactic protein 1 （MCP 1） and macrophage migration inhibition factor （MIF）were detected by ELISA,and the membrane molecule CD16 and CD68 were tested by flow cytometry.THP 1 derived macrophages were pretreated with different concentrations of YXST （10 mg/L,50 mg/L and 100 mg/L）12 h.Then we stimulated these cel s by ox LDL （50 mg/L）for 24 h and detected the secretion of MIF,MCP 1 and the ex-pression of CD16 and CD68 again.Results The ox LDL up regulatet the secretion of MCP 1 and MIF as a dose dependent manner in THP 1 derived macrophages.With the increasing concentration of ox LDL,the secretion of MCP 1 and MIF increased, 50mg/L group reached their secretion peaks after 24h（MCP 1：29.30± 1.48 pg/mL vs.control 5.71± 1.94 pg/mL;MIF：18.67± 0.15 ng/mL vs.control 4.61± 0.40 ng/mL;P〈0.01）.The expression of CD16 and CD68 were varied with different ox LDL concentrations.The mean fluorescence intensity of CD16 and CD68 was significantly reduced in 50mg/L group （CD16：26.9 vs.control 29.8;CD68：22.3 vs. control 25.2）.YXST inhibited the expression of MCP 1 and MIF induced by ox LDL as a dose dependent manner.With the increas-ing concentration of Yangxinshi,the secretion of MCP 1 and MIF decreased.The 100mg/L group declined mostly（MCP 1：9.95± 2.09 pg/mL vs.control 33.30± 2.37 pg/mL;MIF：4.85± 0.12 ng/mL vs.control 18.65± 0.15 ng/mL;P〈0.01）.Yangxinshi change the ex-pression of CD16 and CD68 reduced
出处
《中西医结合心脑血管病杂志》
2016年第15期1722-1726,共5页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
