低氧训练对营养肥胖型大鼠血清外泌体Wnt3a表达的影响

doi:10.19586/j.2095-2341.2023.0136

生物技术进展 ›› 2024, Vol. 14 ›› Issue (4): 685-693.DOI: 10.19586/j.2095-2341.2023.0136

• 研究论文 • 上一篇

低氧训练对营养肥胖型大鼠血清外泌体Wnt3a表达的影响

姜星¹(), 孟昶², 陈龙盼¹, 王清路¹, 田雪文¹()

^1.山东体育学院山东省科学健身指导中心，济南 250102
^2.齐鲁医药学院管理学院，山东淄博 255300

收稿日期:2023-10-23 接受日期:2024-03-27 出版日期:2024-07-25 发布日期:2024-08-07
通讯作者: 田雪文
作者简介:姜星 E-mail：1589106722@qq.com；
基金资助:
国家自然科学基金项目(31701042)

Effects of Hypoxic Exercise on the Expression of Serum Exosome Wnt3a in Obese Rats

Xing JIANG¹(), Chang MENG², Longpan CHEN¹, Qinglu WANG¹, Xuewen TIAN¹()

^1.Shandong Scientific Fitness Guidance Center，Shandong Sport University，Jinan 250102，China
^2.College of Management，Qilu Medical University，Shandong Zibo 255300，China

Received:2023-10-23 Accepted:2024-03-27 Online:2024-07-25 Published:2024-08-07
Contact: Xuewen TIAN

摘要/Abstract

摘要：

目前，肥胖已被公认为一种全球性流行病，研究发现，训练可通过Wnt/β-catenin信号通路降低肥胖大鼠体质量并改善其脂代谢，且Wnt3a在其中发挥着重要的作用，但影响脂肪组织中Wnt3a表达的机制尚不清楚，研究旨在探讨不同环境下训练后营养肥胖型大鼠血清外泌体Wnt3a的表达变化以完善训练影响脂代谢的理论体系。按照随机分配原则将40只肥胖大鼠分为4组，每组10只：常氧训练组（CE）、常氧安静组（C）、低氧训练组（HE）、低氧安静组（H），进行4周干预训练。测量各组大鼠的体质量、肾周脂肪的质量以及体长；通过血脂生化检测盒来检测大鼠血清中低密度脂蛋白胆固醇（low-density lipoprotein cholesterol, LDL-C）、高密度脂蛋白胆固醇（high-density lipoprotein cholesterol, HDL-C）、胆固醇（total cholesterol, TC）以及甘油三酯（triglyceride, TG）的水平；分离血清外泌体并进行蛋白质组学分析。结果发现，低氧训练使肥胖大鼠体质量、脂体比、Lee's指数、TC、TG和LDL-C降低（P＜0.05），HDL-C升高（P＜0.05）；TEM电镜观察结果显示，C组、H组、CE组以及HE组均可以清晰地观察到外泌体胞外囊泡样结构，并在外泌体WB检测中均发现CD9、CD63和TSG101条带，WB检测结果发现，进行单纯低氧或单纯运动Wnt3a蛋白表达情况无显著性变化，低氧环境下Wnt3a蛋白表达水平显著提高。3种干预方式均可改善肥胖大鼠的血脂水平，低氧训练更为显著；低氧训练提升血清外泌体中Wnt3a含量。

关键词: 低氧训练, Wnt3a, 信号通路, 降脂减重

Abstract:

Currently， obesity stands as a worldwide epidemic. Research has found that exercise training could effectively reduce body weight and enhances lipid metabolism in obese rats， with Wnt/β-catenin signaling pathway playing a pivotal role， especially Wnt3a. However， the mechanism underlying of influence Wnt3a in adipose tissue remains unclear. This study aimed to delve into the expression alterations of serum exosome Wnt3a in nutritionally obese rats following exercise training under varying hypoxic conditions， ultimately contributing to the theoretical framework of lipid metabolism modulated by hypoxic training. A total of forty obese rats were randomly assigned into four groups （n=10 each）： normoxic exercise group （CE）， normoxic quiet group （C）， hypoxic exercise group （HE）， and hypoxic quiet group （H）. After a four-week hypoxic exercise intervention， the body weight， perirenal fat weight， and body length were recorded. The serum levels of low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， total cholesterol （TC）， and triglyceride （TG） were determined using a lipid biochemical detection kit. Subsequently， serum exosomes were isolated and subjected to proteomic analysis. The findings revealed that hypoxic training significantly reduced body weight， fat-body ratio， Lee's index， TC， TG， and LDL-C levels （P< 0.05）， while increasing HDL-C （P< 0.05） in obese rats. TEM electron microscopy distinctly depicted the extracellular vesicle-like structure of exosomes in groups C， H， CE， and HE， and WB detection of exosomes confirmed the presence of CD9， CD63 and TSG101 bands. Notably， the WB analysis showed no significant difference in Wnt3a protein expression between simple hypoxia and exercise alone. However， hypoxia significantly elevated Wnt3a protein expression. All three intervention methods contributed to improved blood lipid profiles in obese rats， with hypoxic training exhibiting a more profound effect. Consequently， hypoxic training increased the level of Wnt3a in serum exosomes.

Key words: hypoxia training, Wnt3a, signaling pathway, lipid-lowering weight loss

中图分类号:

R723.14

姜星, 孟昶, 陈龙盼, 王清路, 田雪文. 低氧训练对营养肥胖型大鼠血清外泌体Wnt3a表达的影响[J]. 生物技术进展, 2024, 14(4): 685-693.

Xing JIANG, Chang MENG, Longpan CHEN, Qinglu WANG, Xuewen TIAN. Effects of Hypoxic Exercise on the Expression of Serum Exosome Wnt3a in Obese Rats[J]. Current Biotechnology, 2024, 14(4): 685-693.

图/表 7

图1 实验动物分组及干预流程注：适应训练：CE组速度从12 m·min-1梯度递增到25 m·min-1，运动时间从20 min·d-1递增到60 min·d-1；HE组速度从8 m·min-1梯度递增到20 m·min-1，运动时间从20 min·d-1递增到60 min·d-1。

Fig. 1 Experimental animal grouping and intervention procedure

图2 对照组与肥胖模型组大鼠体质量及血脂指标变化情况A：对照组与肥胖模型组体质量对比；B：对照组与肥胖模型组血脂指标比较情况。*和**表示在P<0.05、P<0.01水平差异有统计学意义。

Fig. 2 Changes in body weight and lipid parameters in the control and obesity model groups

图3 对照组与肥胖模型组大鼠肝脏油红O染色情况

Fig. 3 Oil red O staining of the liver of rats in the control and obesity model groups

图4 各组大鼠脂体比、体质量和Lee's指数变化情况A：实验干预后各组大鼠脂体比变化情况；B：实验干预后各组大鼠体质量和Lee's指数变化情况。*和**表示在P<0.05、P<0.01水平差异有统计学意义。

Fig. 4 Changes in fat-body ratio, body weight and Lee's index in each group

图5 各组大鼠血脂指标变化情况A：实验干预后各组大鼠TC指标变化情况；B：实验干预后各组大鼠TG指标变化情况；C：实验干预后各组大鼠HDL-C指标变化情况；D：实验干预后各组大鼠LDL-C指标变化情况。*和**表示在P<0.05、P<0.01水平差异有统计学意义。

Fig. 5 Changes in lipid parameters in each group

图6 大鼠血清中外泌体表达情况A：利用Wnt3a包被的磁珠筛选外泌体，TEM电镜（100 nm）观察情况；B：实验干预后外泌体WB检测，条带显示情况；C：实验干预后各组大鼠外泌体蛋白质含量检测情况

Fig. 6 Exosome expression in rat serum

图7 大鼠血清外泌体中Wnt3a的表达情况A：对外泌体进行显著富集的 KEGG 通路统计（HE vs. CE），对外泌体蛋白变化影响显著的信号通路进行分析；B：外泌体进行蛋白质组学分析，Wnt3a含量对比情况；C：对Wnt3a阳性外泌体中Wnt3a进行Western blot分析情况。

Fig. 7 Expression of Wnt3a in rat serum exosomes

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低氧训练对营养肥胖型大鼠血清外泌体Wnt3a表达的影响

Effects of Hypoxic Exercise on the Expression of Serum Exosome Wnt3a in Obese Rats

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