应用单细胞测序探索参七解毒颗粒对衰老小鼠模型心肾功能的潜在药效机制  

Exploring the Potential Pharmacological Mechanisms of Shenqi Jiedu Granules on Heart and Kidney Functions in Aging Mice Mouse by Single-Cell Sequencing

作　　者：陈星伶 赵强强 李锦 孙术宁[1,2] 何星灵 张小娇 李思静 李姿儒 林声荣 倪世豪 鲁路 CHEN Xingling;ZHAO Qiangqiang;LI Jin;SUN Shuning;HE Xingling;ZHANG Xiaojiao;LI Sijing;LI Ziru;LIN Shengrong;NI Shihao;LU Lu(National Key Laboratory of Traditional Chinese Medicine Syndrome,The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Guangdong Clinical Research Institute of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院中医证候全国重点实验室,广东广州510405 [2]广州中医药大学岭南医学研究中心,广东广州510405 [3]广东省中医临床研究院,广东广州510405

出　　处：《中药新药与临床药理》2025年第3期435-445,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(82374406);中华中医药学会青年人才托举工程项目(CACM-2021-QNRC2-B30);广东省自然基金青年提升项目(2023A1515030146);广州中医药大学青年菁英人才培养“揭榜挂帅”团队项目;广州中医学院第一临床学院2023年度优秀博士学位论文培养项目(YB202301)。

摘　　要：目的应用单细胞测序技术探索参七解毒颗粒对衰老及未明确潜能的克隆性造血(CHIP)小鼠模型心衰后心肾功能的潜在药效机制。方法将10只C57BL/6J年轻小鼠作为对照组,50只C57BL/6J老年小鼠随机分为模型组、参七解毒颗粒(中药)低浓度组(7.8 g·kg^(-1))、中药中浓度组(15.6 g·kg^(-1))和中药高浓度组(31.2 g·kg^(-1))及达格列净组(10 mg·kg^(-1)混悬液),每组10只,每日给药1次,连续给药10周。30只CD45.1老年小鼠,随机分为对照组、Tet2+/-移植组和Tet2+/-移植+中药组(高浓度),每组10只,进行非清髓型骨髓移植(BMT);8周后,植入式缓释泵皮下注射1.5 mg·kg^(-1)·min-1的血管紧张素Ⅱ,对BMT后的小鼠进行持续注射,对照组小鼠则输注PBS,4周后开始药物治疗。给药10周后,使用小动物超声观察小鼠心功能变化;检测小鼠血肌酐水平;Masson染色法观察小鼠肾脏形态学变化及胶原纤维沉积情况;qPCR法检测小鼠心脏、肾脏炎症和衰老相关基因mRNA表达;免疫荧光法观察小鼠心脏Collagen I、α-SMA表达;通过单细胞转录组数据联合流式细胞术探究参七解毒颗粒改善模型组小鼠心脏、肾脏纤维化的作用和机制。结果与模型组比较,中药高浓度组可改善心脏收缩和舒张功能(P<0.0001),心脏免疫荧光结果显示心脏Collagen I、α-SMA蛋白表达下降(P<0.0001),并有效改善血肌酐水平和肾脏纤维化(P<0.0001),降低炎症和衰老相关基因(IL-6、TNF、P21、S100A8)mRNA表达(P<0.0001);单细胞数据拟时序分析结果发现,与对照组比较,模型组克隆性造血信号明显升高,细胞明显向克隆性造血方向分化;与模型组比较,中药干预组克隆性造血信号明显下降,细胞向CHIP方向减少。qPCR结果表明中药干预的小鼠心脏和肾脏中的炎症因子(TNF)和衰老标记物(Spp1、S100a8)的mRNA水平均明显下降(P<0.05)。Tet2+/-骨髓原代巨噬细胞与成纤维细胞共培养免疫荧光结果显示,中药�Objective To explore the potential pharmacodynamic mechanisms of the Shenqi Jiedu Granules in improving cardiac and renal function in a mouse model of heart failure induced by clonal hematopoiesis of indeterminate potential(CHIP)using single-cell sequencing technology.Methods There were 10 C57BL/6J young mice were used as control group,and 50 C57BL/6J old mice were randomly divided into model group,low-concentration group(7.8 g·kg^(-1)),medium-concentration group(15.6 g·kg^(-1)),high-concentration group(31.2 g·kg^(-1))and Dapagliflozin group(10 mg·kg^(-1)suspension),10 mice in each group,once a day for 10 consecutvie weeks.Thirty CD45.1 aged mice were randomly divided into control group,Tet2+/-transplantation group and Tet2+/-transplantation+Chinese medicine group(high concentration),with 10 mice in each group.Non-myeloablative bone marrow transplantation(BMT)was performed.After 8 weeks,1.5 mg·kg^(-1)·min-1 angiotensin II was injected subcutaneously by osmotic pumps,and the mice after BMT were continuously injected,while the mice in the control group were infused with PBS.Drug treatment was started after 4 weeks.After 10 weeks of administration,the changes of cardiac function in mice were observed by small animal ultrasound.The serum creatinine level of mice was detected;Masson staining was used to observe the morphological changes and collagen fiber deposition in the kidney of mice.The mRNA expressions of inflammation and aging related genes in heart and kidney of mice was detected by qPCR.The expressions of Collagen I and α-SMA in mouse heart was observed by immunofluorescence.The effect and mechanism of Shenqi Jiedu Granuless on improving cardiac and renal fibrosis in model group mice were explored by single cell transcriptome data combined with flow cytometry.Results Compared with the model group,high concentration group of Shenqi Jiedu Granules improved cardiac systolic and diastolic functions(P<0.0001),cardiac immunofluorescence results showed a decrease in the protein expressions of cardiac Collagen

关 键 词：衰老 益气活血解毒 参七解毒颗粒 心肾功能 单细胞测序 克隆性造血 小鼠 

分 类 号：R285.5[医药卫生—中药学]