Chen G, Xiang X, Zeng Z, Huang R, Jin S, Xiao M, Song C
College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China.
Ezhou Central Hospital, Department of Pharmacy, Ezhou 436000, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Jul 20;44(7):1382-1388. doi: 10.12122/j.issn.1673-4254.2024.07.18.
To evaluate the regulatory effect of (DWYG) Decoction on lysoglycerophospholipids (Lyso-GPLs) in circulating exosomes in a mouse model of nonalcoholic fatty liver disease (NAFLD).
Circulating exosomes isolated from mouse serum by size exclusion chromatography were morphologically characterized using transmission electron microscope and examined for surface markers CD9, CD63 and TSG101 using Western blotting. Twenty-four male Kunming mice were randomized into 3 groups for normal feeding (control, =8) or high-fat diet feeding for 1 week to induce NAFLD, after which the latter mice were given DWYG decoction (treatment group, =8) or normal saline (model group, =8) by gavage for 4 weeks. After the last treatment, blood samples were collected from the mice for testing serum TC, HDL-C, LDL-C, ALT and AST levels and isolating circulating exosomes. Using multivariate statistical analysis based on targeted metabolomics strategy, the potential biomarkers for Lyso-GPLs in the exosomes were screened.
The isolated exosomes about 100 nm in size had a typical saucer-like structure with distinct double-layer membranes and a mean particle size of 137.5 nm and expressed the specific surface marker proteins CD9, CD63 and TSG101. The mouse models of NAFLD had significantly increased serum levels of TC, HDL-C, LDL-C and AST and lowered serum ALT level. A total of 43 Lyso-GPLs with significant reduction after DWYG Decoction treatment were identified in NAFLD mice.
DWYG Decoction can regulate Lyso-GPLs in circulating exosomes in NAFLD mice, which provides a new clue for studying the therapeutic mechanism of DWYG Decoction for liver disease.
在非酒精性脂肪性肝病(NAFLD)小鼠模型中,评估(DWYG)汤剂对循环外泌体中溶血甘油磷脂(Lyso - GPLs)的调节作用。
通过尺寸排阻色谱法从小鼠血清中分离循环外泌体,用透射电子显微镜对其形态进行表征,并用蛋白质印迹法检测表面标志物CD9、CD63和TSG101。将24只雄性昆明小鼠随机分为3组,一组正常喂养(对照组,n = 8),另一组高脂饮食喂养1周以诱导NAFLD,之后对后者给予DWYG汤剂(治疗组,n = 8)或生理盐水(模型组,n = 8)灌胃4周。末次处理后,采集小鼠血液样本检测血清总胆固醇(TC)、高密度脂蛋白胆固醇(HDL - C)、低密度脂蛋白胆固醇(LDL - C)、谷丙转氨酶(ALT)和谷草转氨酶(AST)水平,并分离循环外泌体。采用基于靶向代谢组学策略的多变量统计分析,筛选外泌体中Lyso - GPLs的潜在生物标志物。
分离得到的外泌体大小约为100 nm,具有典型的碟状结构,双层膜明显,平均粒径为137.5 nm,表达特异性表面标志物蛋白CD9、CD63和TSG101。NAFLD小鼠模型血清TC、HDL - C、LDL - C和AST水平显著升高,血清ALT水平降低。在NAFLD小鼠中鉴定出43种经DWYG汤剂处理后显著减少的Lyso - GPLs。
DWYG汤剂可调节NAFLD小鼠循环外泌体中的Lyso - GPLs,为研究DWYG汤剂治疗肝病的机制提供了新线索。