基于高通量转录组测序和网络药理学方法筛选抗高尿酸血症燕麦活性肽

Screening of Anti-hyperuricemia Peptides from Oat Protein Based on High-throughput Transcriptome Sequencing and Network Pharmacology

从组学出发为挖掘和筛选抗高尿酸血症燕麦活性肽,并研究燕麦活性肽在抗高尿酸血症上的作用机制。本研究以燕麦籽粒为原料提取RNA,用于高通量转录组测序,通过与参考基因组比对和蛋白编码基因表达量定量以获取燕麦籽粒蛋白序列,利用高通量虚拟酶解、Peptide Ranker和ADME/T筛选燕麦活性肽成分,借助网络药理学技术挖掘燕麦蛋白源抗高尿酸血症活性多肽。结果表明,‘坝莜1号’和‘白燕7号’燕麦籽粒在籽粒形成期和灌浆期所表达重复性低于90%的蛋白序列数分别为6310、3157和5804、5107条。其中最优肽文库为在灌浆期裸燕麦‘坝莜1号’的蛋白序列经蛋白酶K模拟酶解产生的肽文库,在该条件获得的肽文库中预测筛选出42条具有潜在活性和良好成药性的燕麦活性肽。初步筛选主要抗高尿酸血症燕麦活性肽序列为PPF、PPPL、MPF、MPL和PPPF,可能通过靶向ALB、IL1B、SRC、CASP3和STAT3等基因,作用于癌症、脂质与动脉粥样硬化和化学致癌-受体激活等信号通路干预高尿酸血症。分子对接验证显示结合能小于-5 kJ/mol占整体82.86%,表示燕麦多肽主要活性成分与大部分靶点结合活性较好。此外,最优燕麦合成多肽PPF表现出良好的黄嘌呤氧化酶抑制效果(IC_(50)=6.132 mmol/L)。裸燕麦蛋白可作为酶解释放抗高尿酸血症活性肽的良好前体,同时为燕麦蛋白酶解生产生物活性肽和利用燕麦活性肽开发抗高尿酸血症功能性食品提供理论参考。

Initiating from omics,the research aimed to discover and filter oat active peptides effective against hyperuricemia,and to study the operational mechanism of these oat active peptides in treating hyperuricemia.In the present study,RNA was extracted from oat grains for high-throughput transcriptomic sequencing,and oat grain protein sequences were acquired by comparing with a reference genome and quantifying the expression of protein-coding genes.Active components of oat peptides were selected by employing high-throughput enzymolysis in silico,Peptide Ranker,and ADME/T.Network pharmacology were utilized to discover active peptides from oat proteins that were effective against hyperuricemia.The findings indicated that for'Bayou No.1'and'Baiyan No.7'oat grains,the counts of protein sequences expressed with under 90%repeatability during the grain formation and grain-filling phases were respectively 6310,3157,and 5804,5107.The optimal peptide library was from the protein sequences of'Bayou No.1'naked oat in the grain-filling stage,processed through simulated enzymatic digestion in silico with proteinase K,yielding a library with 42 oat active peptides predicted to have potential activity and favorable pharmacological properties.The initial screening revealed key oat active peptides sequences against hyperuricemia to be PPF,PPPL,MPF,MPL,and PPPF,potentially targeting genes like ALB,IL1B,SRC,CASP3,and STAT3,influencing pathways in cancer,lipid and atherosclerosis,and chemically induced carcinogenesis-receptor activation to mitigate hyperuricemia.Molecular docking showed that binding energy<-5 kJ/mol accounted for 82.86%,indicating that the main active components of oat peptides had good binding activity with most of the targets.The optimal oat synthetic peptide PPF showed good xanthine oxidase inhibition(IC_(50)=6.132 mmol/L).Naked oat protein can act as a promising precursor for the enzymatic release of active peptides effective against hyperuricemia,and also offers theoretical guidance for producing bioactive peptides through oat protein enzymolysis and developing functional foods using oat active peptides to combat hyperuricemia.