背景:脂质代谢失调是影响糖尿病肾病(DN)进展的关键因素。莫罗尼甙(MOR)是一种从传统中药山茱萸中分离出的主要活性化合物,我们之前的研究发现,它可以改善肾小管上皮细胞的脂质沉积。本研究旨在探讨MOR是否能改善足细胞脂质沉积及其降低DN的作用机制。
方法:最初,利用网络药理学和生物信息学技术预测MOR与DN的关系。随后,通过分子对接研究了MOR与脂质相关蛋白的结合活性,以确定MOR如何通过这些蛋白发挥作用.在确定MOR的目标之后,进行了动物实验和细胞测试以进行验证。
结果:使用网络药理学,生物信息学,和分子对接,预测和筛选MOR治疗DN的靶蛋白,包括PGC-1α,LXRs,ABCA1,PPARY,CD36和nephrin。特别注意MOR有效结合PGC-1α,而LXR,ABCA1、PPARY和CD36是PGC-1α的下游分子。沉默PGC-1α基因显著降低MOR的治疗效果。相反,在没有PGC-1α敲除的组中,MOR能够增加PGC-1α的表达水平并影响下游蛋白的表达。此外,通过体内和体外实验,利用脂滴染色等技术,PAS,MASSON染色,免疫荧光,和蛋白质印迹,我们发现MOR有效地提高了足细胞蛋白nephrin和脂质代谢调节蛋白PGC-1α的表达水平,PPARY,和ABCA1,同时显著抑制脂质积累启动子CD36的表达。
结论:MOR可通过PGC-1α/LXRs/ABCA1信号通路调节足细胞内胆固醇流出,并通过PGC-1α/PPARY/CD36信号通路控制胆固醇摄入,从而改善DN中的脂质沉积。
BACKGROUND: Dysregulation of lipid metabolism is a key factor influencing the progression of diabetic nephropathy (DN). Morroniside (MOR) is a major active compound isolated from the traditional Chinese herb Cornus officinalis, our previous research found that it can improve the lipid deposition of renal tubular epithelial cells. The purpose of this
study is to explore whether MOR can improve podocyte lipid deposition and its mechanism of reducing DN.
METHODS: Initially, we used network pharmacology and bioinformatics techniques to predict the relationship between renal lipid metabolism of MOR and DN. Subsequently, the binding activity of MOR with lipid-related proteins was studied by molecular docking to determine how MOR acts through these proteins. After determining the target of MOR, animal experiments and cell tests were carried out to verify it.
RESULTS: Using network pharmacology, bioinformatics, and molecular docking, target proteins for MOR treatment of DN were predicted and screened, including PGC-1α, LXRs, ABCA1, PPARY, CD36, and nephrin. It is particularly noted that MOR effectively binds to PGC-1α, while LXRs, ABCA1, PPARY and CD36 are downstream molecules of PGC-1α. Silencing the PGC-1α gene significantly reduced the therapeutic effects of MOR. Conversely, in groups without PGC-1α knockdown, MOR was able to increase the expression levels of PGC-1α and influence the expression of downstream proteins. Furthermore, through in vivo and in vitro experiments, utilizing techniques such as lipid droplet staining, PAS, MASSON staining, immunofluorescence, and Western blot, we found that MOR effectively elevated the expression levels of the podocyte protein nephrin and lipid metabolism-regulating proteins PGC-1α, PPARY, and ABCA1, while significantly inhibiting the expression of the lipid accumulation promoter CD36.
CONCLUSIONS: MOR can regulate the cholesterol efflux in
podocytes via the PGC-1α/LXRs/ABCA1 signaling pathway, and control cholesterol intake via the PGC-1α/PPARY/CD36 signaling pathway, thereby ameliorating lipid deposition in DN.