基于蛋白质组学探讨淫羊藿苷联合迷迭香酸抗慢性肾功能衰竭大鼠肾纤维化的作用机制  

Exploration on mechanism of icariin combined with rosmarinic acid against renal fibrosis in rats with chronic renal failure based on proteomics

作　　者：王静[1] 王凌晨 王蒙 WANG Jing;WANG Lingchen;WANG Meng(Department of Traditional Chinese Medicine,Huadong Hospital Af filiated to Fudan University,Shanghai 200040,China;Department of Nephrology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Traditional Chinese Medicine Institute of Kidney Disease,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)

机构地区：[1]复旦大学附属华东医院中医内科,上海200040 [2]上海中医药大学附属曙光医院肾病科,上海201203 [3]上海中医药大学中医肾病研究所,上海201203 [4]上海中医药大学肝肾疾病病证教育部重点实验室,上海201203 [5]上海市中医临床重点实验室,上海201203

出　　处：《中华中医药杂志》2024年第12期6729-6734,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.82104769);上海市科技创新行动扬帆计划(No.21YF1447900)。

摘　　要：目的:利用蛋白质组学研究淫羊藿苷(ICA)联合迷迭香酸(RA)对慢性肾功能衰竭大鼠肾组织蛋白表达的影响,探讨二者配伍抗肾纤维化(RF)的作用机制。方法:SD雄性大鼠采用5/6(A/I)术制备慢性肾功能衰竭大鼠模型,随机分为模型组、RA组、ICA组、ICA合RA组(IR组)和中药组,每组5只;另取5只大鼠为假手术组。干预8周后,HE和Masson染色比较各组大鼠肾组织病理损伤和小管间质纤维化程度;Western Blot检测纤维连接蛋白(FN)和Ⅰ型胶原(Col-Ⅰ)蛋白表达。另取大鼠肾组织制备蛋白样品,运用串联质谱标签(TMT)标记定量蛋白质组学技术检测IR组较模型组的差异蛋白,并进行生物信息学分析。结果:与假手术组比较,模型组大鼠肾组织出现明显肾小管间质损伤和胶原沉积,FN和Col-Ⅰ蛋白表达显著升高(P<0.01)。与模型组比较,各药物组肾小管间质损伤和间质纤维化明显减轻,FN和Col-Ⅰ蛋白表达均降低(除RA组FN蛋白外)(P<0.05,P<0.01),尤以IR组和中药组较为明显。与模型组比较,IR组大鼠肾组织中有258个差异蛋白,其中上调蛋白有41个,下调蛋白有217个。GO富集分析显示,差异蛋白主要富集于对雌二醇的反应、炎症反应的调节、对氧化应激反应的调节等生物过程,主要富集于载氧活性、氧结合、NADH结合、转录因子结合等分子功能。KEGG分析显示,差异蛋白所涉及的上调信号通路主要有丙酮酸代谢、PPAR信号通路、泛酸和CoA的生物合成、柠檬酸循环等通路,所涉及的下调信号通路主要有局部粘连、铁死亡、ECM受体相互作用、紧密连接等通路。结论:利用蛋白质组学筛选的差异蛋白及涉及的分子功能、生物过程和信号通路可能是ICA和RA配伍抗慢性肾功能衰竭RF的作用机制,具有深入研究的价值。Objective:To investigate the effect of icariin(ICA)combined with rosmarinic acid(RA)on the protein expression in the kidney tissue of rats with chronic renal failure(CRF)using the proteomics approach,and to explore the underlying mechanism of their combination against renal fibrosis(RF).Methods:The rat model of CRF was induced in male SD rats with 5/6 ablation and infarction(A/I)surgery,the model rats were randomly divided into the following groups:model group,RA group,ICA group,ICA combined with RA group(IR group)and Chinese medicine group,5 rats in each group.Another 5 rats were taken as sham-operated group.After 8 weeks of treatment,HE and Masson staining were performed to compare the degree of histopathological damage and tubulointerstitial fibrosis in each group of rats,and the expression of fibronectin(FN)and collagen-Ⅰ(Col-Ⅰ)proteins was detected by Western Blot.Protein samples were also prepared from rat kidney tissues to detect differential proteins in the IR group compared to the model group using TMT-labeled quantitative proteomics technology and bioinformatics analysis was performed.Results:Compared with the sham-operated group,the kidney tissues of the model group showed obvious tubular interstitial injury and collagen deposition,and the expression of FN and Col-Ⅰproteins was significantly increased(P<0.01).Compared with the model group,renal tubular interstitial injury and interstitial fibrosis were significantly reduced in each drug group,and the expression of FN and Col-Ⅰproteins was decreased(except for FN protein expression in the RA group)(P<0.05,P<0.01),especially the IR group and the Chinese medicine group.A total of 258 differential proteins in the kidney tissue of the IR group rats compared to the model group,including 41 up-regulated proteins and 217 down-regulated proteins.GO analysis showed that the differential proteins were mainly enriched in biological processes(BP)such as response to estradiol,regulation of inflammatory response,regulation of response to oxidative stress,and

关 键 词：蛋白质组学 肾纤维化 淫羊藿苷 迷迭香酸 信号通路 机制 慢性肾功能衰竭 生物信息学 

分 类 号：R28[医药卫生—中药学]