艾塞那肽对糖尿病小鼠下丘脑氧化应激的保护作用及其机制

doi:10.3969/j.issn.1006-5725.2025.03.004

摘要/Abstract

摘要：

目的探究艾塞那肽（Exe）对糖尿病小鼠下丘脑氧化应激的影响及其可能机制。方法将C57BL/6J小鼠适应性喂养1周后随机分为CON组（普通饲料）、T2DM组（高脂饲料）及T2DM+Exe组（高脂饲料+艾塞那肽），T2DM+Exe组高脂喂养8周后，予腹腔内注射艾塞那肽［24 nmol/（kg·d）］8周。检测小鼠体质量及糖脂代谢指标，酶联免疫吸附法（ELISA）检测小鼠炎症因子及脂肪因子水平，Western blot检测下丘脑黑皮素4受体（MC4R）及前阿黑皮素原（POMC）表达，提取下丘脑线粒体，采用流式细胞仪检测线粒体活性氧（ROS）含量，试剂盒检测线粒体丙二醛（MDA）含量、超氧化物歧化酶（SOD）活性，透射电镜观察线粒体超微结构的变化。体外实验采用棕榈酸（PA）和艾塞那肽干预下丘脑GT1-7细胞，并应用短发夹RNA（shRNA）沉默MC4R，观察细胞氧化应激和脂质沉积情况。结果与CON组相比，T2DM组小鼠糖脂代谢指标、促炎因子及脂肪因子水平明显升高（P < 0.05），下丘脑MC4R及POMC蛋白表达减少（P < 0.05），下丘脑线粒体ROS及MDA含量显著增加（P < 0.05），而SOD活性下降（P < 0.05），线粒体形态异常。艾塞那肽干预后，上述指标均得到显著改善。体外实验抑制MC4R表达后，与艾塞那肽干预组相比，ROS及MDA含量显著增加（P < 0.05），SOD活性降低（P < 0.05），细胞内出现脂质沉积。结论艾塞那肽对糖尿病小鼠下丘脑氧化应激损伤具有一定的保护作用，其机制与上调下丘脑MC4R表达有关。

关键词: 艾塞那肽, 糖尿病, 下丘脑氧化应激, 黑皮素受体4

Abstract:

Objective To explore the effect of exenatide on oxidative stress in the hypothalamus of diabetes mice and its potential mechanism. Methods After one week of adaptive feeding， C57BL/6J mice were randomly divided into the CON group （normal chaw diet）， the T2DM group （high-fat diet， HFD）， and the T2DM+Exe group （HFD+ exenatide）. After 8 weeks of HFD， mice in the T2DM+Exe group were intraperitoneally injected with exenatide ［24 nmol/（kg·d）］ for 8 weeks.The weight and glucose and lipid metabolism levels of the mice were measured， and the levels of inflammatory and adipokine factors in mice were detected using the ELISA method. Western Blot was used to detect the expression of melanocortin receptor-4 （MC4R） and proopiomelanocortin （POMC） in the hypothalamus. Hypothalamic mitochondria were extracted， and the content of mitochondrial reactive oxygen species （ROS） was measured using a flow cytometer. The content of malondialdehyde （MDA） and the activities of superoxide dismutase （SOD） in the mitochondria were detected using assay kits. Changes in the ultrastructure of mitochondria were observed using a transmission electron microscope. In vitro experiments， palmitic acid （PA） and exenatide were used to treat hypothalamic GT1-7 cells， and short hairpin RNA （shRNA） was used to silence the melanocortin 4 receptor （MC4R）， and observe the cellular oxidative stress and lipid deposition. Results Compared with the CON group， the T2DM group mice showed a significant increase in glucose and lipid metabolism indicators， pro-inflammatory factors， and adipose factor levels （P < 0.05）， the expression of MC4R and POMC proteins in the hypothalamus were decreased （P < 0.05）， and the mitochondrial ROS and MDA content in the hypothalamus significantly were increased （P < 0.05）， while SOD and CAT activities were decreased （P < 0.05）. Mitochondrial morphology was abnormal. After intervention with exenatide， the above indicators were significantly improved. After inhibiting MC4R expression in vitro experiments， compared with the intervention group with exenatide， the ROS and MDA content was significantly increased （P < 0.05）， SOD activity was decreased （P < 0.05）， and lipid deposition occurred in the cells. Conclusions Exenatide exhibits a protective effect on hypothalamic oxidative stress injury in diabetic mice， and this mechanism may be associated with the upregulation of MC4R expression.

Key words: exenatide, diabetes mellitus, hypothalamic oxidative stress, melanocortin receptor-4

中图分类号:

R587.1

郑路,张好好,吴飞飞,郭佳琪,王有琴,郝芮敏,冯李慧,李燕. 艾塞那肽对糖尿病小鼠下丘脑氧化应激的保护作用及其机制[J]. 实用医学杂志, 2025, 41(3): 330-338.

Lu ZHENG,Haohao ZHANG,Feifei WU,Jiaqi GUO,Youqin WANG,Ruimin HAO,Lihui FENG,Yan. LI. Protective effect of exenatide on oxidative stress in hypothalamus of diabetes mice and its mechanism[J]. The Journal of Practical Medicine, 2025, 41(3): 330-338.

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组别	体质量/g	FBG/（mmol/L）	TC/（mmol/L）	TG/（mmol/L）	FFA/（mmol/L）	Ins/（mIU/L）	HOMA-IR
NC组	30.35 ± 3.34	5.70 ± 0.93	2.36 ± 0.72	0.50 ± 1.17	0.41 ± 0.12	2.51 ± 0.12	0.64 ± 0.13
T2DM组	35.32 ± 2.65^?	14.15 ± 2.24^?	4.99 ± 0.39^?	1.73 ± 0.24^?	0.83 ± 0.08^?	5.56 ± 0.45^?	3.52 ± 0.78^?
T2DM+Exe组	31.45 ± 2.00^#	9.50 ± 0.85^?#	3.97 ± 0.28^?#	0.88 ± 0.10^?#	0.34 ± 0.14^#	2.65 ± 0.13^#	1.12 ± 0.10^?#
F值	10.33	32.45	34.99	61.67	25.93	153.34	44.83
P值	< 0.05	< 0.05	< 0.05	< 0.05	< 0.05	< 0.05	< 0.05

组别	IL-4/ （pg/mL）	TNF-α/ （pg/mL）	LEP/ （pg/mL）	Visfitin/ （ng/mL）
NC组	185.74 ± 7.75	1.04 ± 0.79	0.76 ± 0.08	17.75 ± 0.72
T2DM组	102.13 ± 16.29^?	17.71 ± 2.38^?	6.06 ± 0.24^?	28.98 ± 1.09^?
T2DM+Exe组	169.04 ± 3.44^?#	2.25 ± 1.02^#	0.96 ± 0.12^#	19.66 ± 0.79^*#
F值	87.014	176.09	1764.931	233.229
P值	< 0.05	< 0.05	< 0.05	< 0.05

组别	MC4R蛋白水平	POMC蛋白水平
NC组	1.00 ± 0.13	1.00 ± 0.11
T2DM组	0.50 ± 0.11^?	0.53 ± 0.10^?
T2DM+Exe组	0.75 ± 0.10^?#	0.80 ± 0.10^?#
F值	27.613	32.647
P值	< 0.05	< 0.05

组别	ROS/（MFI）	MDA/（nmol/mgprot）	SOD/（U/mgprot）
NC组	1.00 ± 0.13	3.22 ± 0.40	13.42 ± 2.30
T2DM组	2.15 ± 0.39^?	6.56 ± 1.98^?	4.94 ± 1.50^?
T2DM+Exe组	1.24 ± 0.37^#	4.35 ± 1.85^#	7.80 ± 1.32^?#
F值	21.726	11.041	35.8
P值	< 0.05	< 0.05	< 0.05

组别	MC4R mRNA
CON组	1.03 ± 0.30
MC4R shRNA1组	0.25 ± 0.06^?
MC4R shRNA2组	0.35 ± 0.03^?*
MC4R shRNA3组	0.15 ± 0.02^?
F值	18.397
P值	< 0.05