络风宁1号方对ox-LDL介导的人树突状细胞成熟过程中IL-27/JAK-STATs信号通路的影响  

Influence of Luofengning Granules I on IL-27/JAK-STATs signaling pathway during ox-LDLmediated maturation of human dendritic cells

作　　者：魏蔚 孙晓婷 孔维颖 李海燕[1] 王显[1] Wei Wei;Sun Xiaoting;Kong Weiying;Li Haiyan;Wang Xian(Department of Cardiology,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;不详)

机构地区：[1]北京中医药大学东直门医院心内科,北京100700 [2]郑州市骨科医院综合内科,郑州450052 [3]北京市密云区中医医院内一科,北京101500

出　　处：《中国循证心血管医学杂志》2020年第9期1109-1113,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

摘　　要：目的探究络风宁1号方对氧化低密度脂蛋白(ox-LDL)介导的人树突状细胞(DCs)成熟过程中白介素27/Janus激酶信号转导子和转录激活子(IL-27/JAK-STATs)信号通路的影响。方法选取临床确诊为冠状动脉粥样硬化性心脏病(冠心病)的患者,采集其外周血样本,用密度梯度离心法分离出单个核细胞(PBMCs),用磁珠分选出CD14+单核细胞,并诱导成未成熟的DCs。选取10只大鼠,随机分为空白组和药物组,每组5只,分别以络风宁1号方和等量生理盐水灌胃,7 d后腹主动脉取血,取血清备用。将未成熟的DCs分为3组:空白组,模型组,药物组。空白组加入空白大鼠血清,模型组加入空白大鼠血清和ox-LDL,药物组加入含药大鼠血清和ox-LDL,培养48 h。酶联免疫吸附测定(ELISA)检测细胞上清液中IL-27水平,蛋白质印迹法(Western blot)检测白细胞分化抗原36(CD36)、凝集素样氧化低密度脂蛋白受体-1(LOX-1)、Janus激酶1(JAK1)、Janus激酶2(JAK2)、信号转导子和转录激活子1(STAT1)、信号转导子和转录激活子3(STAT3)蛋白水平。结果模型组的IL-27、JAK1、JAK2、STAT1、STAT3、CD36、LOX-1较空白组均升高,差异具有统计学意义(P<0.05)。药物组的IL-27、JAK1、JAK2、STAT1、STAT3、CD36、LOX-1水平较模型组均下降,差异具有统计学意义(P<0.05)。结论ox-LDL促进外周血来源的未成熟DCs分泌IL-27,上调JAK1、JAK2、STAT1、STAT3蛋白表达,上调与ox-LDL结合的受体CD36、LOX1的表达,而络风宁1号方含药血清则可以下调上述蛋白的表达,从而抑制ox-LDL介导的DCs免疫成熟过程。这可能是络风宁1号方抗动脉粥样硬化的机制之一。Objective To investigate the influence of Luofengning Granules I on interleukin-27(IL-27)/JAK-STATs signaling pathway during the maturation of human dendritic cells(DCs)mediated by oxidized low density lipoprotein(ox-LDL).Methods The patients with coronary heart disease(CHD)were chosen and samples of peripheral blood were collected from them.The peripheral blood mononuclear cells were isolated by using density gradient centrifugation,and CD14+monocytes were separated by using magnetic beads to induce immature DCs.The rats(n=10)were divided randomly into blank group and medication group(each n=5),and respectively given orally normal saline and Luofengning Granules I.After 7 d,the serum samples were collected from abdominal aortic blood.The immature DCs were divided into blank group,model group and medication group.The blank group was given blank rat serum,model group was given blank rat serum and ox-LDL,and medication group was given medicated rat serum and ox-LDL and all groups were cultivated for 48 h.The level of IL-27 was detected in supernatant by using ELISA.The levels of CD36,LOX-1,JAK1,JAK2,STAT1 and STAT3 were detected by using Western blotting assay.Results The levels of IL-27,JAK1,JAK2,STAT1,STAT3,CD36 and LOX-1 increased in model group than those in blank group(P<0.05).The levels of IL-27,JAK1,JAK2,STAT1,STAT3,CD36 and LOX-1 decreased in medication group than those in model group(P<0.05).Conclusion Ox-LDL can improve peripheral blood immature DCs to secrete IL-27,up-regulate the protein expressions of JAK1,JAK2,STAT1 and STAT3,and up-regulate the expressions of receptors CD36 and LOX-1 bound to ox-LDL.Luofengning Granules I can down-regulate all above protein expressions and inhibit DCs maturation mediated by ox-LDL,which may be one of mechanisms of Luofengning Granules I’s anti-atherosclerosis effect.

关 键 词：络风宁1号方 动脉粥样硬化 树突状细胞 白介素27 JAK-STATs信号通路 大鼠 

分 类 号：R543.5[医药卫生—心血管疾病]