基于网络药理学和实验验证探究软坚清脉颗粒治疗动脉硬化闭塞症的作用机制  

Exploration on the mechanism of Ruanjian Qingmai Granules on the treatment of arteriosclerosis obliterans based on network pharmacology and experimental validation

作　　者：邓婕 洪碧莹 蔡同凯 李嘉诚 曹烨民 贾成林 曹永兵 DENG Jie;HONG Biying;CAI Tongkai;LI Jiacheng;CAO Yemin;JIA Chenglin;CAO Yongbing(Institute of Vascular Disease,Shanghai TCM-Integrated Hospital,Shanghai University of TCM,Shanghai 200082,China;School of Basic Medicine and Clinical Pharmacy,China Pharmaceutical University,Nanjing 210009,China;Shanghai Diacart Biomedical Science and Technology Limited Company,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属上海市中西医结合医院脉管病研究所,上海200082 [2]中国药科大学基础医学与临床药学学院,南京210009 [3]上海迪亚凯特生物医药科技有限公司,上海201203

出　　处：《中华中医药杂志》2024年第12期6762-6768,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家科技重大专项重大新药创制(No.2019ZX09201004-002-092);上海市卫生健康委员会卫生行业临床研究专项基金(No.202140334);上海中医药大学预算内项目(No.2021LK062)。

摘　　要：目的:运用网络药理学探索软坚清脉颗粒(RJQM)治疗动脉硬化闭塞症(ASO)的潜在治疗靶点及机制,并利用载脂蛋白E基因缺陷(ApoE^(-/-))小鼠验证。方法:利用数据库分析RJQM作用于ASO的交集靶点,进行GO、KEGG信号通路分析。30只ApoE^(-/-)小鼠高脂饮食,随机分为模型组,RJQM低、中、高剂量组及阳性药组,每组6只。以同周龄C57BL/6J小鼠作为空白对照组。连续灌胃18周。测定血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平,HE染色检测肝脏、主动脉弓形态学变化,油红O染色观察主动脉弓根部的病变程度,并结合qRT-PCR检测肝脏中ABCA1、ABCG5、ABCG8mRNA表达情况。结果:KEGG通路富集分析发现RJQM可能通过调节脂质代谢信号通路发挥治疗作用。与模型组比较,RJQM高剂量组TG水平显著降低(P<0.05);RJQM各剂量组肝脏细胞肿胀与脂肪空泡样得到改善,主动脉弓中粥样斑块面积显著减小(P<0.01),油红着色脂滴面积显著减小(P<0.05);RJQM高剂量组肝脏中ABCA1、ABCG5 mRNA表达显著上调(P<0.01);RJQM各剂量组ABCG8 mRNA表达显著下调(P<0.01)。结论:RJQM能够调控脂质和动脉硬化信号通路,通过影响脂质代谢途径发挥干预ASO的作用。Objective:To explore the potential therapeutic targets and mechanism of Ruanjian Qingmai Granules(RJQM)on the treatment of arteriosclerosis obliterans(ASO)by using network pharmacology techniques,and further validated using ApoE^(-/-)mice.Methods:Various databases were used to analyze the intersectional targets of RJQM action on ASO,and GO and KEGG signaling pathway analysis was performed.A total of 30 ApoE^(-/-)mice were given a high-fat diet,and randomly divided into model group,RJQM low-dose group(RJQM-L),RJQM medium-dose group(RJQM-M),RJQM high-dose group(RJQM-H),and positive control group,6 mice in each group,C57BL/6J mice of the same weekly age were used as control group.These mice were gavaged continuously for 18 weeks.The serum levels of TC,TG,LDL-C and HDL-C were measured.The morphological changes of liver tissue and aortic arch were detected by HE staining.The lipid deposition in intact arteries of mice was detected by oil red ostaining.And the mRNA expression of ABCA1,ABCG5,and ABCG8 genes related to cholesterol metabolism was detected by qRT-PCR.Results:KEGG pathway enrichment analysis revealed that RJQM may play a therapeutic role by regulating lipid metabolism signaling pathway.Compared with the model group,the TG level was reduced in the RJQM-H group(P<0.05),the swelling of liver cells and fat vacuolation pattern in mice treated with three dose RJQM groups were improved,the area of atheromatous plaque in the aortic arch significantly reduced(P<0.01),and the area of oil-red coloring lipid droplets significantly reduced(P<0.05),the expression of ABCA1 and ABCG5 mRNA was significantly up-regulated in the RJQM-H group(P<0.01),and the expression of ABCG8 mRNA in three dose RJQM groups was down-regulated(P<0.01).Conclusions:RJQM can regulate lipid and atherosclerotic signaling pathways and exert an intervention in ASO.

关 键 词：网络药理学 软坚清脉颗粒 动脉硬化闭塞症 ApoE^(-/-)小鼠 胆固醇代谢 脂质代谢 机制 

分 类 号：R54[医药卫生—心血管疾病]