白术多糖通过花生四烯酸代谢通路缓解环磷酰胺诱导的小鼠肾损伤

李冰心; 龚淑影; 许丹宁; 李婉雁; 曹楠; 付新亮; 江丹莉; 洪龙胜; 余梓榆; 白帅飞; 田允波

doi:10.13386/j.issn1002-0306.2023070030

白术多糖通过花生四烯酸代谢通路缓解环磷酰胺诱导的小鼠肾损伤

Polysaccharide of Atractylodes macrocephala Koidz Alleviate Kidney Injury Induced by Cyclophosphamide in Mice through Arachidonic Acid Metabolic Pathway

摘要

摘要: 本研究旨在探讨白术多糖（PAMK）对环磷酰胺（CTX）诱导的小鼠肾脏损伤的影响及潜在的作用机制。选用100只42~43日龄雄性C57BL/6小鼠随机分为四组，白术多糖（PAMK）组和白术多糖+环磷酰胺（PAMK+CTX）组灌胃200 mg/kg PAMK，1次/d；对照（Control）组和环磷酰胺（CTX）组给予等量的生理盐水。实验第25~27 d，CTX组和PAMK+CTX组腹腔注射100 mg/kg CTX，1次/d；Control组和PAMK组注射等量生理盐水。实验第35 d采集肾脏进行组织学观察，氧化应激检测和转录组测序。结果显示，与CTX组相比，PAMK+CTX组肾脏损伤有所缓解；肾脏中丙二醛量显著下降（P<0.05）、谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶活性和总抗氧化能力显著升高（P<0.05）。为了进一步探究PAMK缓解CTX诱导肾损伤的调控机制，本实验进行了肾脏转录组测序。结果显示，在Control组vs CTX组和CTX组vs PAMK+CTX组分别鉴定到493个和333个差异表达基因（DEGs）。两组DEGs功能富集分析结果发现DEGs富集在花生四烯酸代谢等信号通路。花生四烯酸通路相关基因检测结果显示，与Control组相比，CTX组Cyp2b9 、PTGS1、NF-κB和Mfsd2a mRNA表达量显著升高（P<0.05）；与CTX组相比，PAMK+CTX组Cyp2c65 、BCL6 mRNA表达量显著升高（P<0.05），Cyp2b9 、PTGS1 和Mfsd2a mRNA表达量显著降低（P<0.05）。综上所述，PAMK可能通过花生四烯酸代谢通路缓解小鼠肾脏氧化应激，从而降低CTX诱导的小鼠肾脏损伤。

Abstract: This study aimed to investigate the effects of the polysaccharide of Atractylodes macrocephala Koidz (PAMK) on cyclophosphamide (CTX)-induced renal injury in mice and its potential underlying mechanisms. One hundred male C57BL/6 mice, aged 42~43 days, were randomly divided into four groups, with five repetitions in each group and five mice in each repetition. The PAMK group and PAMK+CTX group were orally administered 200 mg/kg PAMK once daily, while the control group and CTX group were given an equivalent amount of saline. From days 25~27 of the experiment, the CTX group and PAMK+CTX group were intraperitoneally injected with 100 mg/kg CTX once daily, while the control group and PAMK group were injected with an equivalent amount of saline. On day 35 of the experiment, the kidneys were collected for histological observation, oxidative stress detection, and transcriptome sequencing. The results showed that compared with the CTX group, the renal injury in the PAMK+CTX group was alleviated. The content of malondialdehyde (MDA) in the kidneys was significantly decreased (P<0.05), while the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), and total antioxidant capacity (T-AOC) were significantly increased (P<0.05). To further explore the regulatory mechanisms of PAMK in alleviating CTX-induced renal injury, transcriptome sequencing of the kidneys was performed. The results showed that compared with the control group, 493 differentially expressed genes (DEGs) were identified in the CTX group vs control group comparison, and 333 DEGs were identified in the CTX group vs PAMK+CTX group comparison. Functional enrichment analysis of the DEGs in both groups revealed significant enrichment in signaling pathways related to arachidonic acid metabolism. The expression levels of arachidonic acid pathway-related genes were examined, and it was found that compared with the control group, the mRNA expression levels of Cyp2b9, PTGS1, NF-κB and Mfsd2a were significantly increased in the CTX group (P<0.05). On the other hand, in the PAMK+CTX group, the expression levels of Cyp2c65 and BCL6 were significantly increased (P<0.05), while the expression levels of Cyp2b9, PTGS1and Mfsd2a were significantly decreased (P<0.05). In conclusion, PAMK may alleviate oxidative stress in the kidneys of mice and reduce CTX-induced renal injury through the arachidonic acid metabolism pathway.