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PPAR α、PGC-1α、Acox1的蛋白表达,下调了 TGF-β1、Cαspase3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.PPAR α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.
Abstract
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-1of CsA.The AS group was given 4.2,8.4 g·kg-1·d-1,respectively,while the Val group was given 10 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 PPAR α,PGC-1 α,Acoxl,and downregulated TGF-β 1 and Caspase3.It improved renal tissue apoptosis,lipid accumulation,and type 1 and type 3 collagen deposition.After PPAR α knockdown,the protein expression of PGC-1 α and Acox1 was downregulated,and TGF-β 1 and Caspase3 was upregulated,and renal tissue apoptosis,lipid accumulation,and type 1 and type 3 collagen deposition were intensified.After intervention with AS,there were varying degrees of improvement.Conclusion:AS can regulate PPAR α/PGC-1 α/TGF-β 1 axis to improve fatty acid oxidation,reduce renal tissue apoptosis,lipid accumulation,and collagen deposition,and improve inflammation,oxidative stress and lipid overload,and protect renal function.
基金项目
国家自然科学基金青年科学基金项目(82204886)
中国博士后科学基金特别资助(2023T160398)
中国博士后科学基金面上项目(2021M702039)
山东省自然科学基金青年基金项目(ZR2022QH133)
国家科技期刊平台
NSTL
万方数据