暖心康调控CD72hi巨噬细胞对压力负荷致心力衰竭小鼠的作用和有效成分  

Effects and active component of Nuanxinkang on regulating CD72hi macrophages to improve heart failure mice induced by pressure load

作　　者：倪世豪 孙术宁 李悦[1,2,3,4] 黎欢 王陵军 冼绍祥[1,2,3,4] 鲁路 杨忠奇[1,2,3,4] NI Shi-hao;SUN Shu-ning;LI Yue;LI Huan;WANG Lngjnun;XIAN Shao-xiang;LU Lu;YANG Zhong-qi(The First Afiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Guangdong Provincial University Key Laboratory of Traditional Chinese Medicine Prevention and Treatment of Chronic Heart Failure,Guangzhou 510405,China;Guangzhou Key Laboratory for Chinese Medicine Prevention and Treatment of Chronic Heart Failure,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一附属医院,广州510405 [2]广州中医药大学岭南医学研究中心,广州510405 [3]广东省普通高校慢性心力衰竭中医药防治重点实验室,广州510405 [4]广州市慢性心力衰竭中医药防治重点实验室,广州510405

出　　处：《中华中医药杂志》2023年第5期2329-2335,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家中医临床研究基地建设项目(No.国中医药科技函[2018]131号);2022年医疗卫生健康事业发展专项资金(传承发展中医药事业)中医药人才培养类项目(No.08002011009);国家重点研发计划(No.2018YFC1707401);国家自然科学基金项目(No.81803928);青年人才托举工程项目(No.2021-QNRC2-B30);广东省自然科学基金项目(No.2021A1515011457);广州市科技计划项目重点研发计划(No.202206080015);广州市科技计划(No.202102020269)。

摘　　要：目的:探究暖心康通过CD72hi巨噬细胞(CD72hi-CMφs)治疗慢性心力衰竭小鼠的作用和有效成分。方法:采用主动脉弓缩窄术(TAC)建立慢性心力衰竭小鼠模型,治疗6周后综合评估暖心康对TAC小鼠心脏和炎症的影响。使用单细胞测序标记物检测多种心力衰竭病理和心力衰竭小鼠模型中新型炎症亚群CD72hi-CMφs信号,通过转录组联合反卷积算法评估暖心康对CD72hi-CMφs的影响,并基于关键靶点筛选暖心康调控CD72hiCMφs的有效成分。通过体外研究暖心康有效成分对CD72hi-CMφs分化以及相关细胞因子的影响。结果:与模型组比较,暖心康改善心力衰竭小鼠模型心功能和心室重构,减少炎症。单细胞数据结果显示CD72hi-CMφs在各种心力衰竭病理和心力衰竭动物模型中均有显著升高(P<0.01)。生物信息分析和实验结果显示暖心康降低CD72hiCMφs水平(P<0.01),且调控CD72hi-CMφs的转录因子Rel。分子对接结果显示暖心康有效成分果糖精氨酸能与c-Rel活性区域结合。荧光素酶报告基因检测显示果糖精氨酸减少了由CD72启动子Rel驱动的荧光素酶活性(P<0.01)。细胞实验显示果糖精氨酸能够抑制CD72hi-CMφs分化及相关炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的表达(P<0.01)。结论:暖心康可能通过果糖精氨酸抑制CD72hi-CMφs改善慢性心力衰竭。Objective:To explore the effects and active component of Nuanxinkang(NXK)on mouse chronic heart failure model via regulating CD72hi macrophages(CD72hi-CMφs).Methods:Heart failure was induced by transverse aortic constriction.After a six-week-treatment,the effects of NXK on the mice's hearts and cardiac inflammation was evaluated.Single-cell sequencing markers were used to detect CD72hi-CMφs signatures in several heart failure pathologies and animal models.A transcriptome joint deconvolution algorithm was applied to explore how NXK impacted CD72hi-CMφs.Based on the critical target,the active component of NXK regulating CD72hi CMφs was screened,with its effect on CD72hi-CMφs differentation and related cytokines studied.Results:NXK improved cardiac function,ameliorated ventricular remodeling and reduced infammation.CD72hi-CMφs were significantly upregulated in various heart failure pathologies and animal models(P<0.01).In the bioinformatical analysis and experimental validation,NXK inhibited CD72hi-CMφs expression(P<0.01)and regulated CD72hi-CMφs transcription factor Rel.Molecular docking identified fructose arginine(FA),an active component of NXK,can bind to the c-Rel active region.Luciferase reporter assay showed that FA reduced lucifrase activity driven Rel by the CD72 promoter(P<0.01).FA also inhibited CD72hi-CMφs differentiation and related infammatory factors TNF-a,IL-IβmRNA expression in cell experiments(P<0.01).Conclusion:NXK may inhibit CD72hi-CMφs through FA to improve chronic heart failure.

关 键 词：慢性心力衰竭 单细胞测序 CD72hi巨噬细胞 暖心康 果糖精氨酸 

分 类 号：R285.5[医药卫生—中药学]