宁心痛颗粒及其有效组分抑制HSP60诱导的THP-1细胞炎症反应的实验研究  

Inhibitory Effects of Ningxintong Granule and its Effective Components on the THP-1 Cells Inflammation Induced by Extracellular Human Heat Shock Protein 60

作　　者：高颖[1] 陈蕊[2] 顾宁[2] 何小丽[2] GAO Ying;CHEN Rui;GU Ning;HE Xiao-li(Postdoctoral Mobile Research Station of Nanjing University of Chinese Medi-cine,Nanjing（210046;Department of Cardiology,the Third Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing（210001)

机构地区：[1]南京中医药大学博士后流动站,南京210046 [2]南京中医药大学第三附属医院心血管科,南京210001

出　　处：《中国中西医结合杂志》2018年第9期1087-1092,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(No.81173399);中国博士后科学基金项目(No.2015M571791);江苏省博士后科学基金项目(No.1401137C);江苏省自然科学基金面上项目(No.BK20161115);十三五南京市医学科技创新平台重大项目(No.ZDX16013)

摘　　要：目的观察NF-κB、MAPKs信号通路在介导重组人热休克蛋白60(HSP60)诱导巨噬细胞(Mφ)炎症反应中的作用,探讨益气活血方宁心痛颗粒的抗炎机制。方法采用人急性单核白血病细胞株(human acute monocytic leukemia cell line,THP-1)体外培养模型,结合血清药理学与中药组分配伍方法进行论证。将THP-1细胞诱导分化为Mφ,分为对照组(Mφ+培养液)、HSP60(10μg/mL)组、黄芪多糖(APS,200μg/mL)组、藁本内酯(LIG,20μg/mL)组、APS(200μg/mL)+LIG(20μg/mL)组、含药血清(3%)组、辛伐他汀(15μg/mL)组,各药物干预组以相应治疗药物预处理24h后,除对照组外,其余各组均加入终浓度为10μg/mL的HSP60。检测磷酸化的NF-κB(p-NF-κB)、MAPKs[包括p-p38、胞外信号调节激酶1/2(p-ERK1/2)和c-Jun氨基末端激酶(p-JNK)]蛋白表达及TNF-α、IL-6含量。结果与对照组比较,HSP60组的p-NF-κB及p-p38、p-JNK、p-ERK1/2蛋白表达和TNF-α、IL-6含量升高(P<0.01);与HSP60组比较,各药物干预组检测蛋白表达和TNF-α、IL-6含量均降低(P<0.05,P<0.01);与APS组比较,APS+LIG组、含药血清组和辛伐他汀组检测蛋白表达和TNF-α、IL-6含量均降低(P<0.05,P<0.01);与LIG组比较,APS+LIG组检测蛋白表达均降低(P<0.05,P<0.01),TNF-α、IL-6含量差异无统计学意义(P>0.05),含药血清组和辛伐他汀组检测蛋白表达及TNF-α、IL-6含量均降低(P<0.05,P<0.01);含药血清组和辛伐他汀组均较APS+LIG组蛋白表达和TNF-α、IL-6含量降低更明显(P<0.05,P<0.01),且辛伐他汀组降低蛋白表达的效果优于含药血清组(P<0.01)。结论 NF-κB及MAPKs信号通路是HSP60诱导THP-1源性Mφ发生炎症反应的关键信号通路,宁心痛颗粒含药血清、APS、LIG、APS配伍LIG可不同程度通过调控以上通路发挥抗炎效果,其中宁心痛颗粒含药血清的效果最强。Objective To explore the inhibitory effects of Ningxintong Granule（NXT）, which can invigorate qi and activate blood, and its effective components on the inflammatory reaction induced by extracellular recombinated heat shock protein 60 （HSP60） in the human acute monocytic leukemia cell line（THP-1 ） and the underlying anti-inflammatory mechanisms. Methods The THP-1 were cultured in vitro and the serum drug of NXT was used in coordination with component combination of Astragalus membranceus -Ligusticum wallichii herb.The THP-1 cells were differentiated into macrophages （Mψ） and then divided into the control group,the HSP60 group,the astragalus polysaccharide（APS） group, the li- gustilide（LIG） group,the APS ＋ LIG group, the NXT group and the simvastatin group. HSP60 with ultimateconcentration of 10 μg/mL was added into all but the control group after incubation of APS （200 μg/mL）, LIG （20 μg/mL）, APS（200 μg/mL） plus LIG（20 μg/mL）, NXT（3%） or simvastatin （15 μg/mL） for 24 h, respectively. The protein expressions of phosphate-nuclear factor kappa B（NF-κB） and mitogen actived protein kinases [ including p38, c-Jun NH2-terminal kinases （JNK）, extracellular signal-regulated kinasel/2 （ERK112） ] and the contents of TNF-α and IL-6 were detected. Results Com pared with the control group, the expressions of the four proteins and the contents of TNF-α and IL-6 in the HSP60 group were all ele- vated remarkably（P 〈0.01 ）, which were decreased significantly in the APS group, the LIG group, the APS ＋LIG group, the NXT group and the simvastatin group（P 〈0.05, P 〈0.01 ）. Compared with the APS group, those in the APS ＋ LIG group, the NXT group and the simvastatin group were decreased dramatic- ly（P 〈0.05, P 〈0.01 ）. In contrast to the LIG group, the four protein expressions were decreased out- standingly（P 〈0.05, P 〈0.01 ） in the APS ＋ LIG group, while there was not statistically significant in the contents of TNF-α and IL-6 b

关 键 词：宁心痛颗粒 核因子-κB 丝裂原活化蛋白激酶 热休克蛋白60 

分 类 号：R285.5[医药卫生—中药学]