耐力运动联合间歇性冷刺激促进大鼠白色脂肪组织棕色化

王潇; 李梁; 崔成立; 蔡志平; 于畅; 张立佳

doi:10.16098/j.issn.0529-1356.2023.03.014

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解剖学报 ›› 2023, Vol. 54 ›› Issue (3) : 348-356. DOI: 10.16098/j.issn.0529-1356.2023.03.014

组织学胚胎学发育生物学

耐力运动联合间歇性冷刺激促进大鼠白色脂肪组织棕色化

王潇¹ 李梁^1* 崔成立¹ 蔡志平¹ 于畅² 张立佳²

作者信息 +

Endurance exercise combined with intermittent cold stimulation promoting white adipose tissue browning in rats

WANG Xiao¹ LI Liang^1* CUI Cheng-li¹ CAI Zhi-ping¹ YU Chang² ZHANG Li-jia²

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文章历史 +

摘要

目的探讨耐力运动和间歇性冷刺激对肥胖大鼠棕色脂肪组织活性及白色脂肪组织棕色化的影响及机制。方法 52只SD雄性肥胖大鼠分为常温对照组（NC组）、间歇性冷刺激组（IC组）、耐力运动组（EE组）和耐力运动联合间歇性冷刺激组（EI组），每组13只，常温对照组环境温度为24~26℃；运动组采用动物跑台运动，跑速为20 m/min，坡度为0°，每次运动50 min，运动2次，中间间歇20 min；冷刺激组在环境温度为4 ℃下暴露4 h（低温仓内），每周刺激6 d，持续4周；持续刺激4周后，正电子发射断层显像/X 线计算机体层成像（PET/CT）检测大鼠肩胛区棕色脂肪组织（BAT）代谢情况及腹股沟区白色脂肪组织（iWAT）棕色化情况；称体重、量体长、取材，采集血样检测血脂4项；免疫组织化学和Western blotting检测大鼠BAT和iWAT线粒体解耦联蛋白1(UCP1)、血管内皮生长因子A (VEGF-A)、p38 MAPK、过氧化物酶体增殖物激活受体-γ共激活因子-1α(PGC-1α)蛋白表达。结果 IC组、EE组、EI组大鼠体重及血清总胆固醇（CHOL）、甘油三酯（TG）、低密度脂蛋白（LDL）显著降低。PET/CT显示，在BAT中，IC组、EE组、EI组最大标准摄取值（SUVmax）显著升高；在iWAT中，EI组SUVmax显著升高。免疫组织化学显示，IC组、EE组、EI组脂肪细胞的数目增多，密度增高，iWAT 脂肪细胞中呈现棕色脂肪样细胞。Western blotting显示，在BAT中，IC组、EE组、EI组UCP1 蛋白表达水平显著升高；在iWAT中，IC组、EE组、EI组UCP1、PGC-1α 蛋白表达水平明显升高；IC组、EI组p38 MAPK蛋白表达水平明显升高；EE组、EI组VEGF-A蛋白表达水平明显降低。结论 3种实验方案干预后均能促进肥胖大鼠棕色脂肪组织活化并且促进白色脂肪组织棕色化，以间歇性冷刺激联合耐力运动作用最为显著。

Abstract

Objective To explore the effect and mechanism of endurance exercise and intermittent cold stimulation on brown adipose tissue activity and white adipose tissue browning in obese rats. Methods Totally 52 SD male obese rats were divided into normothermic control group (NC group)，intermittent cold stimulation group (IC group)，endurance exercise group (EE group) and endurance exercise combined with intermittent cold stimulation group (EI group),13 pieces per group. The ambient temperature of the normothermic control group was 24-26℃；The exercise group used animal running table exercise with a running speed of 20 m/min and a slope of 0°，50 minutes per exercise，exercise 2 times with 20 minutes interval; The cold stimulation group was exposed to an ambient temperature of 4℃ for 4 hours （in the cold storage）, and stimulated 6 days per week for 4 weeks. After 4 weeks of continuous stimulation, positron emission tomography/computed tomography (PET/CT) was applied to detect the metabolism of interscapular brown adipose tissue (BAT) and browning of inguinal white adipose tissue (iWAT) in rats. Weighting，measuring body length，sampling tissue and collecting blood samples were detected the four lipids. The expression of mitochondrial uncoupling protein 1(UCP1)，vascular endothelial growth factor-A(VEGF-A)，p38 MAPK and peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1α) in interscapular brown adipose tissue and inguinal white adipose tissue of rats were detected by immunohistochemistry and Western blotting. Results The body weight，serum total cholesterol （CHOL），triglyceride（TG）and low density lipoprotein （LDL）of IC，EE and EI groups decreased significantly. PET/CT showed that, in BAT，SUVmax increased significantly in IC，EE and EI groups；In iWAT，maximum standard intake value (SUVmax) increased significantly in EI group. Immunohistochemistry showed: in BAT and iWAT, the number and the density of adipocytes of IC, EE and EI groups increased，and iWAT adipocytes appeared brown adipocytes. Western blotting showed that, in the BAT, the expression level of UCP1 protein in IC，EE and EI groups increased significantly；In the iWAT, the expression levels of UCP1 and PGC-1α in IC group，EE group and EI group increased significantly ；The expression level of p38 MAPK protein in IC group and EI group increased significantly；The expression level of VEGF-A protein in EE group and EI group decreased significantly. Conclusion All three experimental protocols can promote brown adipose tissue activation and white adipose tissue browning in obese rats after intervention，with intermittent cold stimulation combined with endurance exercise having the most significant effect.

关键词

耐力运动 / 冷刺激 / 白色脂肪组织 / 棕色脂肪组织 / 免疫组织化学 / 免疫印迹法 / 正电子发射断层显像/X 线计算机体层成像 / 大鼠

Key words

Endurance exercise

/ Cold stimulation / White adipose tissue / Brown adipose tissue / Immunohistochemistry / Western blotting / Positron emission tomography/computedtomography / Rat

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王潇李梁崔成立蔡志平于畅张立佳. 耐力运动联合间歇性冷刺激促进大鼠白色脂肪组织棕色化[J]. 解剖学报. 2023, 54(3): 348-356 https://doi.org/10.16098/j.issn.0529-1356.2023.03.014

WANG Xiao LI Liang CUI Cheng-li CAI Zhi-ping YU Chang ZHANG Li-jia. Endurance exercise combined with intermittent cold stimulation promoting white adipose tissue browning in rats[J]. Acta Anatomica Sinica. 2023, 54(3): 348-356 https://doi.org/10.16098/j.issn.0529-1356.2023.03.014

中图分类号： R322

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