血必净通过上调ADAM17酶活性抑制EA.hy926细胞TLR4信号通路  

Xuebijing injection inhibits TLR4 signaling pathway through up-regulating ADAM17 activity in EA.hy926 cells

作　　者：张英杰 王晶[2] 齐鸿飞 康雅隆 董静[2] 于远望[1] 王海芳 ZHANG Yingjie;WANG Jing;QI Hongfei;KANG Yalong;DONG Jing;YU Yuanwang;WANG Haifang(Shaanxi Province and Xianyang Key Laboratory of Integrative Traditional Chinese and Western Medicine for the Prevention and Treatment of Cardiovascular Diseases,Shaanxi University of Chinese Medicine,Xi'an 712000,China;Department of Cardiology,Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xi'an 712000,China)

机构地区：[1]陕西中医药大学陕西省及咸阳市中西医结合心血管病防治重点实验室,712000 [2]陕西中医药大学第二附属医院心血管内科,712000

出　　处：《免疫学杂志》2024年第4期359-364,共6页Immunological Journal

基　　金：陕西省感染与免疫疾病重点实验室开放课题(2022-ZD-3);2022年大学生创新创业训练计划(S202210716072);陕西省中医药传承创新暨秦药开发重点项目(2021-02-ZZ-011);陕西中医药大学校内课题(2023GP01)。

摘　　要：目的探索血必净注射液(XBJ)抑制脂多糖(LPS)诱导血管内皮细胞Toll样受体4(TLR4)炎性信号通路的分子机制。方法培养EA.hy926血管内皮细胞,加入不同浓度XBJ处理细胞4 h后,添加100 ng/ml LPS刺激细胞,Western blot法检测p-p38、p-p65、p-ERK、p-JNK及IκBα表达水平。无血清培养EA.hy926细胞过夜,加入20μl/ml XBJ处理细胞不同时间,浓缩培养液,Western blot法检测培养液中TLR4受体含量和脱落酶ADAM17磷酸化水平;利用酶活性试剂盒检测细胞中ADAM17酶活性。结果XBJ显著抑制细胞中TLR4介导的炎性信号。XBJ诱导细胞中p-ADAM17水平和酶活性随时间而升高,同时培养液中TLR4受体含量随时间而增多。TAPI-1(10μmol/L)可显著抑制XBJ诱导的细胞膜TLR4脱落。结论XBJ通过上调ADAM17活性诱导细胞膜TLR4脱落,从而抑制其对细胞内炎性信号的激活。This study was performed to investigate the inhibitory effects of Xuebijing injection(XBJ)on lipopolysaccharide(LPS)-induced inflammatory signals on EA.hy926 vascular endothelial cells and the underlying mechanism,and to provide a theoretical basis for the treatment of sepsis with XBJ.WST-1 assay was used to detect the effects of XBJ on the cell viability;Western blot analysis was performed to detect the protein expression levels of IκBα,p-p65,p-ERK,p-JNK,p-p38,p-AMAD17 in cell lysates and the content of sTLR4 fragment in the concentrated culture supernatants.ADAM17 sheddase activity in cells was detected by using a commercial available kit.Data showed that all of the TLR4-mediated inflammatory signals were significantly inhibited by the treatment of XBJ(P<0.01).ADAM17 phosphorylation and shedding activity were induced by XBJ treatment,simultaneously the sTLR4 contents in the culture media were increased.XBJ-induced shedding of TLR4 was suppressed by the preteatment of 10μmol/L TAPI-1(an ADAM17 inhibitor).Taken together,XBJ can induce the shedding of TLR4 from cell membrane by up-regulating ADAM17 shedding activity,thereby inhibiting the activation of TLR4-mediated intracellular inflammatory signals in EA.hy926 cells.

关 键 词：血必净 血管内皮细胞 炎症 TLR4 ADAM17 

分 类 号：R364.5[医药卫生—病理学] R392.5[医药卫生—基础医学]