该研究旨在阐明黄蜀葵花总黄酮(total flavones of Abelmoschus manihot,TFA)联合恩格列净(empagliflozin,EM)通过调控线粒体稳态和泛凋亡减轻糖尿病肾小管病(diabetic tubulopathy,DT)的作用机制和分子靶点。在体内研究中,...
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该研究旨在阐明黄蜀葵花总黄酮(total flavones of Abelmoschus manihot,TFA)联合恩格列净(empagliflozin,EM)通过调控线粒体稳态和泛凋亡减轻糖尿病肾小管病(diabetic tubulopathy,DT)的作用机制和分子靶点。在体内研究中,首先通过“单肾切除、链脲佐菌素腹腔注射和高脂饲料喂养”建立DT大鼠模型。造模成功后,DT模型鼠分别经灌胃接受TFA、EM、TFA+EM以及生理盐水(安慰剂)连续8周的干预;在体外研究中,使用高糖(high glucose,HG)或分别联合TFA、EM、TFA+EM处理NRK52E细胞或敲低Z-DNA结合蛋白1(Z-DNA binding protein 1,ZBP1)的NRK52E细胞。在体内、外研究中分别观察了肾小管损伤和活性氧(reactive oxygen species,ROS)产物诱导的肾小管上皮细胞损伤的相关特征,分析了肾小管泛凋亡和ZBP1介导的肾小管上皮细胞泛凋亡相关特征,比较了肾脏线粒体裂变标志性分子蛋白表达水平和肾小管上皮细胞线粒体稳态相关特征。此外,在网络药理学研究中,采用HERB数据库和SwissTargetPrediction数据库分别预测并筛选TFA和EM的作用靶点;借助Cytoscape 3.7.2网络图像化软件对筛选出的TFA、EM化学成分及其作用靶点进行相关网络构建;采用OMIM数据库、GeneCards数据库整合DT相关靶点;采用DAVID数据库针对3者交集靶点富集分析基因本体论(Gene Ontology,GO)、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路。体内研究结果显示,TFA+EM能改善DT模型鼠肾小管损伤、肾脏泛凋亡通路关键信号分子蛋白表达水平和表达特征以及肾脏线粒体裂变标志性分子蛋白表达水平,而且,TFA+EM在体内的改善作用均优于TFA或EM;网络药理学的研究结果显示,TFA+EM干预DT可能是通过调控“泛凋亡信号通路”发挥作用;体外研究结果显示,TFA+EM 在HG环境下能改善ROS产物诱导的肾小管上皮细胞损伤、ZBP1介导的泛凋亡以及线粒体稳态,包括线粒体裂变标志性分子蛋白表达水平、线粒体超微结构、膜电位水平,而且,TFA+EM在体外的改善作用均优于TFA或EM。更为重要的是,借助ZBP1敲低的NRK52E细胞,发现在HG环境下,ZBP1的确是介导肾小管上皮细胞发生泛凋亡的上游因素,而TFA+EM能靶向ZBP1调控泛凋亡通路标志性信号分子蛋白表达水平。总之,该研究阐明,TFA+EM不同于TFA或EM,它是通过改善线粒体稳态并抑制ZBP1介导的泛凋亡多靶点减轻DT。这些发现为在DT临床治疗领域使用TFA+EM这一“中西医协同”新策略提供了明确的药理学证据。
骨质疏松症(OP)在绝经后女性和老年男性中较为常见,是全球范围内极为普遍的一种慢性疾病。其主要特征是骨量呈下降趋势、骨骼的脆性不断提高以及骨微结构遭到破坏。外泌体包含了蛋白质、脂质、核酸等多种生物活性物质,能够在细胞间传递信息、调节生理功能,并在多种生理和病理过程中发挥重要作用。在其中,作为功能性组成部分的微小核糖核酸(miRNA)可通过对骨形成与骨吸收过程进行调控,进而参与骨质疏松症的发生和发展进程。虽然在中医典籍中没有明确关于骨质疏松症的记载,但是如今越来越多的人运用中医药来干预骨质疏松的进展。中医药具有副作用较小的优点,并且相关的体内外实验发现,中医药是通过调节外泌体miRNA来发挥作用的。本文阐述外泌体miRNA与骨质疏松骨组织更新和修复中的关系,以及总结中药如何通过外泌体miRNA对OP发挥作用。Osteoporosis (OP) is relatively common in postmenopausal women and elderly men and is an extremely common chronic disease worldwide. Its main characteristics are a downward trend in bone mass, increasing brittleness of bones, and destruction of bone microstructure. Exosomes contain various bioactive substances, such as proteins, lipids, and nucleic acids. They can transmit information between cells, regulate physiological functions, and play an important role in various physiological and pathological processes. Among them, microRNAs (miRNAs), as functional components, can regulate the processes of bone formation and bone resorption and thus participate in the occurrence and development of osteoporosis. Although there is no clear record of osteoporosis in traditional Chinese medicine classics, nowadays more and more people use traditional Chinese medicine to intervene in the progress of osteoporosis. Traditional Chinese medicine has the advantage of relatively small side effects. And relevant in vitro and in vivo experiments have found that traditional Chinese medicine exerts its effects by regulating exosomal miRNAs. This article elaborates on the relationship between exosomal miRNAs and the renewal and repair of osteoporotic bone tissue, and summarizes how traditional Chinese medicine exerts its effects on OP through exosomal miRNAs.
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