黄芪-丹参调控脂肪酸代谢改善环孢素A慢性肾毒性的机制  

Mechanism of Astragalus and Salvia miltiorrhiza on cyclosporine A chronic nephrotoxicity via fatty acid metabolism

作　　者：韩聪 高冉冉[2] 连梦慧 申振 王振国 李伟[1] HAN Cong;GAO Ranran;LIAN Menghui;SHEN Zhen;WANG Zhenguo;LI Wei(Nephropathy Department,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China;Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]山东中医药大学附属医院肾病科,济南250014 [2]山东中医药大学,济南250355

出　　处：《中华中医药杂志》2024年第8期3983-3988,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.82204886);中国博士后科学基金特别资助(No.2023T160398);中国博士后科学基金面上项目(No.2021M702039);山东省自然科学基金青年基金项目(No.ZR2022QH133)。

摘　　要：目的:探讨黄芪-丹参通过调控脂肪酸代谢改善环孢素A(CsA)慢性肾毒性的作用。方法:40只雄性C57BL/6小鼠随机分为5组:正常组、模型组、黄芪-丹参低剂量组、黄芪-丹参高剂量组及缬沙坦组,每组8只。以CsA 30 mg·kg^(-1)·d^(-1)灌胃建立CsA慢性肾毒性模型,造模时黄芪-丹参低、高剂量组组分别给予4.2、8.4 g·kg^(-1)·d^(-1)灌胃,缬沙坦组给予10 mg·kg^(-1)·d^(-1)。6周后观察小鼠肾功能相关血尿指标,HE、Masson及PAS染色观察肾脏病理,肾组织蛋白质测序构建黄芪-丹参干预下的肾脏蛋白表达图谱,并对高度富集的脂肪酸代谢相关通路及差异基因PPARα、GK、Hmgcs2、Plin2、Degs1、Agpat5、Neu1、Pck1进行筛选及验证。结果:与正常组比较,模型组体质量下降、尿量较少,BUN、Scr、UACR、NAG、IL-6、MDA、FFA升高,黄芪-丹参干预后均显著降低(P<0.05,P<0.01)。与正常组比较,模型组肾小管胞质空泡化、扩张及萎缩,肾间质炎症细胞浸润,肾小球基底膜增厚,胶原纤维明显增生,黄芪-丹参干预后均有不同程度改善。鉴定出PPAR信号通路等10条与脂肪酸代谢等密切相关的靶通路,脂肪酸代谢相关通路中的差异蛋白PPARα、GK、Hmgcs2、Plin2、Degs1、Agpat5、Neu1、Pck1均有较高的AUC预测性。与模型组比较,黄芪-丹参上调了PPARα、PGC-1α、Acox1的蛋白表达,下调了TGF-β1、Caspase3的蛋白表达,改善了肾组织凋亡、脂质累积及Ⅰ型与Ⅲ型胶原沉积。PPARα敲减后,PGC-1α、Acox1蛋白表达下调,TGF-β1、Caspase3蛋白表达上调,肾组织凋亡、脂质累积及Ⅰ型与Ⅲ型胶原沉积加剧,黄芪-丹参干预后均有不同程度的改善。结论:黄芪-丹参能够通过调控PPARα/PGC-1α/TGF-β1轴,改善脂肪酸氧化供能,减少肾组织凋亡、脂质累积及胶原沉积,改善机体炎症、氧化应激及脂质过载,保护肾功能。Objective:To explore the improvement effect of Astragalus and Salvia miltiorrhiza (AS) on cyclosporine A(CsA) chronic nephrotoxicity by regulating fatty acid metabolism.Methods:Forty male C57BL/6 mice were randomly divided into 5 groups:Qi deficiency and blood stasis normal group (N group),model group (M group),low-dose AS group (AS-L group),high-dose AS group (AS-H group),and valsartan group (Val group).A chronic nephrotoxicity model of CsA was established by gavage with 30 mg·kg^(-1)·d^(-1) of CsA.The AS group was given 4.2,8.4 g·kg^(-1)·d^(-1),respectively,while the Val group was given10 mg·kg^(-1)·d^(-1).After 6 weeks,renal function related indicators were observed,and renal pathology was observed using HE,Masson,and PAS staining.Renal tissue protein sequencing was performed to construct a renal protein expression map under the intervention of AS,and highly enriched fatty acid metabolism related pathways and differential genes PPARα,GK,Hmgcs2,Plin2,Degs1,Agpat5,Neu1,Pck1 were verified.Results:Compared with N group,the M group had a decrease in body mass,less urine output,and an increase in BUN,Scr,UACR,NAG,IL-6,MDA,and FFA.After intervention with AS,there was a varying degree of decrease (P<0.05,P<0.01).Compared with N group,the M group showed vacuolization,dilation,and atrophy of renal tubular cytoplasm,as well as infiltration of inflammatory cells in the renal interstitium.The basement membrane of the glomerulus thickened,and collagen fibers significantly proliferated.After intervention with AS,there were varying degrees of improvement.Ten target pathways closely related to fatty acid metabolism,such as PPAR signaling pathway,had been identified,and the differential protein PPARα,GK,Hmgcs2,Plin2,Degs1,Agpat5,Neu1,Pck1 all had high predictive power for AUC.Compared with M group,AS upregulated the protein expression of PPARα,PGC-1α,Acox1,and downregulated TGF-β1 and Caspase3.It improved renal tissue apoptosis,lipid accumulation,and type 1 and type 3 collagen deposition.After PPARα knockdown,the pr

关 键 词：蛋白质测序 黄芪-丹参 脂肪酸代谢 环孢素A慢性肾毒性 

分 类 号：R285.5[医药卫生—中药学]