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

Abstract

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

CLC Number:

R587.1

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.

Figures/Tables 11

Tab.1

Fig.1

Tab.2

Tab.3

Fig.2

Tab.4

Fig.3

Fig.4

Fig.5

Tab.5

Tab.6

References 26

1	NCD Risk Factor Collaboration （NCD-RisC）.Worldwide trends in diabetes prevalence and treatment from 1990 to 2022： A pooled analysis of 1108 population-representative studies with 141 million participants ［J］. Lancet， 2024， 404（10467）： 2077-2093.
2	PANG D， YANG C， LUO Q， et al. Soy isoflavones improve the oxidative stress induced hypothalamic inflammation and apoptosis in high fat diet-induced obese male mice through PGC1-alpha pathway ［J］. Aging （Albany NY）， 2020， 12（9）： 8710-8727. doi:10.18632/aging.103197 doi: 10.18632/aging.103197
3	JO D， YOON G， SONG J. Role of Exendin-4 in Brain Insulin Resistance， Mitochondrial Function， and Neurite Outgrowth in Neurons under Palmitic Acid-Induced Oxidative Stress ［J］. Antioxidants （Basel）， 2021， 10（1）：78. doi:10.3390/antiox10010078 doi: 10.3390/antiox10010078
4	MALONE J I， HANSEN B C. Does obesity cause type 2 diabetes mellitus （T2DM）？Or is it the opposite？［J］. Pediatr Diabetes， 2019， 20（1）： 5-9. doi:10.1111/pedi.12787 doi: 10.1111/pedi.12787
5	马刚，孙家忠. GLP-1 RAs联合恩格列净对2型糖尿病患者的治疗效果及对胰岛素抵抗的影响［J］. 实用医学杂志，2020，36（18）：2500-2504.
6	DE SOUZA CORDEIRO L M， ELSHEIKH A， DEVISETTY N， et al. Hypothalamic MC4R regulates glucose homeostasis through adrenaline-mediated control of glucose reabsorption via renal GLUT2 in mice ［J］. Diabetologia， 2021， 64（1）： 181-194. doi:10.1007/s00125-020-05289-z doi: 10.1007/s00125-020-05289-z
7	POLENI P E， AKIEDA-ASAI S， KODA S， et al. Possible involvement of melanocortin-4-receptor and AMP-activated protein kinase in the interaction of glucagon-like peptide-1 and leptin on feeding in rats ［J］. Biochem Biophys Res Commun， 2012， 420（1）： 36-41. doi:10.1016/j.bbrc.2012.02.109 doi: 10.1016/j.bbrc.2012.02.109
8	TANAKA T， MASUZAKI H， YASUE S， et al. Central melanocortin signaling restores skeletal muscle AMP-activated protein kinase phosphorylation in mice fed a high-fat diet ［J］. Cell Metab， 2007， 5（5）： 395-402. doi:10.1016/j.cmet.2007.04.004 doi: 10.1016/j.cmet.2007.04.004
9	GOKUL P R， APPERLEY L， PARKINSON J， et al. Semaglutide， A Long-Acting GLP-1 Analogue， for the management of early onset obesity due to MC4R defect-A case report ［J］. Horm Res Paediatr， 2024. doi： 10.1159/000537921 . doi: 10.1159/000537921
10	LAMBADIARI V， PAVLIDIS G， KOUSATHANA F， et al. Effects of 6-month treatment with the glucagon like peptide-1 analogue liraglutide on arterial stiffness， left ventricular myocardial deformation and oxidative stress in subjects with newly diagnosed type 2 diabetes ［J］. Cardiovasc Diabetol， 2018， 17（1）： 8. doi:10.1186/s12933-017-0646-z doi: 10.1186/s12933-017-0646-z
11	YAGISHITA Y， URUNO A， FUKUTOMI T， et al. Nrf2 Improves Leptin and Insulin Resistance Provoked by Hypothalamic Oxidative Stress ［J］. Cell Rep， 2017， 18（8）： 2030-2044. doi:10.1016/j.celrep.2017.01.064 doi: 10.1016/j.celrep.2017.01.064
12	LIN M H， CHENG P C， HSIAO P J， et al. The GLP-1 receptor agonist exenatide ameliorates neuroinflammation， locomotor activity， and anxiety-like behavior in mice with diet-induced obesity through the modulation of microglial M2 polarization and downregulation of SR-A4 ［J］. Int Immunopharmacol， 2023， 115： 109653. doi:10.1016/j.intimp.2022.109653 doi: 10.1016/j.intimp.2022.109653
13	STENLID R， CERENIUS S Y， WEN Q， et al. Adolescents with obesity treated with exenatide maintain endogenous GLP-1， reduce DPP-4， and improve glycemic control ［J］. Front Endocrinol （Lausanne）， 2023， 14： 1293093. doi:10.3389/fendo.2023.1293093 doi: 10.3389/fendo.2023.1293093
14	BAI C， WANG Y， NIU Z， et al. Exenatide improves hepatocyte insulin resistance induced by different regional adipose tissue［J］. Front Endocrinol （Lausanne）， 2022， 13： 1012904. doi:10.3389/fendo.2022.1012904 doi: 10.3389/fendo.2022.1012904
15	XU Q， ZHANG X， LI T， et al. Exenatide regulates Th17/Treg balance via PI3K/Akt/FoxO1 pathway in db/db mice ［J］. Mol Med， 2022， 28（1）： 144. doi:10.1186/s10020-022-00574-6 doi: 10.1186/s10020-022-00574-6
16	XU F， CAO H， CHEN Z， et al. Short-term GLP-1 receptor agonist exenatide ameliorates intramyocellular lipid deposition without weight loss in ob/ob mice ［J］. Int J Obes （Lond）， 2020， 44（4）： 937-947. doi:10.1038/s41366-019-0513-y doi: 10.1038/s41366-019-0513-y
17	ADRIAENSSENS A E， BIGGS E K， DARWISH T， et al. Glucose-Dependent Insulinotropic Polypeptide Receptor-Expressing Cells in the Hypothalamus Regulate Food Intake ［J］. Cell Metab， 2019， 30（5）：987-996.e6. doi:10.1016/j.cmet.2019.07.013 doi: 10.1016/j.cmet.2019.07.013
18	HUANG Z， LIU L， ZHANG J， et al. Glucose-sensing glucagon-like peptide-1 receptor neurons in the dorsomedial hypothalamus regulate glucose metabolism ［J］. Sci Adv， 2022， 8（23）： eabn5345. doi:10.1126/sciadv.abn5345 doi: 10.1126/sciadv.abn5345
19	HAVEL P J， HAHN T M， SINDELAR D K， et al. Effects of streptozotocin-induced diabetes and insulin treatment on the hypothalamic melanocortin system and muscle uncoupling protein 3 expression in rats ［J］. Diabetes， 2000， 49（2）： 244-252. doi:10.2337/diabetes.49.2.244 doi: 10.2337/diabetes.49.2.244
20	DONG Y， CARTY J， GOLDSTEIN N， et al. Time and metabolic state-dependent effects of GLP-1R agonists on NPY/AgRP and POMC neuronal activity in vivo ［J］. Mol Metab， 2021， 54： 101352. doi:10.1016/j.molmet.2021.101352 doi: 10.1016/j.molmet.2021.101352
21	SPEZANI R， MARINHO T S， REIS T S， et al. Cotadutide （GLP-1/Glucagon dual receptor agonist） modulates hypothalamic orexigenic and anorexigenic neuropeptides in obese mice ［J］. Peptides， 2024， 173： 171138. doi:10.1016/j.peptides.2023.171138 doi: 10.1016/j.peptides.2023.171138
22	GONZÁLEZ P， LOZANO P， ROS G， et al. Hyperglycemia and Oxidative Stress： An Integral， Updated and Critical Overview of Their Metabolic Interconnections ［J］. Int J Mol Sci， 2023， 24（11）：9352. doi:10.3390/ijms24119352 doi: 10.3390/ijms24119352
23	XIA B， DING J， LI Q， et al. Loganin protects against myocardial ischemia-reperfusion injury by modulating oxidative stress and cellular apoptosis via activation of JAK2/STAT3 signaling ［J］. Int J Cardiol， 2024， 395： 131426. doi:10.1016/j.ijcard.2023.131426 doi: 10.1016/j.ijcard.2023.131426
24	PANDEY S， MANGMOOL S， MADREITER-SOKOLOWSKI C T， et al. Exendin-4 protects against high glucose-induced mitochondrial dysfunction and oxidative stress in SH-SY5Y neuroblastoma cells through GLP-1 receptor/Epac/Akt signaling ［J］. Eur J Pharmacol， 2023， 954： 175896. doi:10.1016/j.ejphar.2023.175896 doi: 10.1016/j.ejphar.2023.175896
25	WANG Y， FANG N， WANG Y， et al. Activating MC4R Promotes Functional Recovery by Repressing Oxidative Stress-Mediated AIM2 Activation Post-spinal Cord Injury ［J］. Mol Neurobiol， 2024， 61（8）：6101-6118. doi:10.1007/s12035-024-03936-9 doi: 10.1007/s12035-024-03936-9
26	ZHANG H H， LIU J， QIN G J， et al. Melanocortin 4 Receptor Activation Attenuates Mitochondrial Dysfunction in Skeletal Muscle of Diabetic Rats ［J］. J Cell Biochem， 2017， 118（11）： 4072-4079. doi:10.1002/jcb.26062 doi: 10.1002/jcb.26062

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

Protective effect of exenatide on oxidative stress in hypothalamus of diabetes mice and its mechanism

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 11

References 26

Related Articles 15

Recommended Articles

Metrics

Comments

组别	ROS/ （MFI）	MDA/ （nmol/mgprot）	SOD/ （U/10⁴cell）
CON组	1.00 ± 0.05	9.64 ± 1.41	0.0118 ± 0.001
PA+sh-NC组	1.54 ± 0.13^?	17.24 ± 1.01^?	0.0079 ± 0.0006^?
PA+Exe+sh-NC组	1.03 ± 0.06^#	10.26 ± 1.21^#	0.010 ± 0.0007^?#
PA+Exe+MC4RshRNA组	1.33 ± 0.15^?&	13.46 ± 0.72^?&	0.009 ± 0.0007^?
F值	29.264	48.683	12.147
P值	< 0.05	< 0.05	< 0.05

[1]	Wenjing ZHENG,Xiangling CHU,Yuqiong WU,Min ZHANG,Xiaohong CHU,Nan ZHANG,Honglin HU. Changes in serum NOV/CCN3 levels in mid⁃ to late⁃term pregnant women and their association with gestational diabetes mellitus and pregnancy outcome [J]. The Journal of Practical Medicine, 2025, 41(1): 71-77.
[2]	Yan LI,Qi WANG,Lu ZHENG,Lihui FENG,Li MA,Jian WEI,Liangshu. LIU. Correlation between IL-17 gene polymorphism and type 2 diabetes mellitus in Han population in southeast Shanxi Province [J]. The Journal of Practical Medicine, 2024, 40(5): 695-701.
[3]	Jianmei SHI,Xixi WANG,Xiaojie WEI. Role of ferritinophagy in the pathogenesis of diabetes and its complications： A review of literature [J]. The Journal of Practical Medicine, 2024, 40(3): 417-422.
[4]	Wenqing XU,Yishan LI,Qiuyu HAN,Fangjing SONG,Lin. MENG. Expression levels of HIF⁃3α methylation and DDIT4 in gestational diabetes mellitus and its relationship with pregnancy outcomes [J]. The Journal of Practical Medicine, 2024, 40(24): 3497-3502.
[5]	Menglu ZHU,Fengjiao ZHANG,Zhiqiang KANG. Effects of telomere length and plasma AGEs on bone mineral density in type 2 diabetic patients [J]. The Journal of Practical Medicine, 2024, 40(20): 2860-2866.
[6]	Chunmei ZHANG,Xinhui HUANG,Jinqiu HU,Xiaoyan BI,Fuli. YA. Sulforaphane protects human platelets from high glucose⁃induced cellular apoptosis through down-regulating PI3K/Akt signaling pathway [J]. The Journal of Practical Medicine, 2024, 40(18): 2530-2536.
[7]	Liehua LIU,Yanbing. LI. Remission of type 2 diabetes： Contending perspectives and practical approaches [J]. The Journal of Practical Medicine, 2024, 40(16): 2206-2210.
[8]	Xin HUANG,Pu ZHANG,Yu GAO,Kai CHEN,Xiaofeng LI,Huiyang GU,Xue. LIANG. Multivariate analysis and prediction model of mild cognitive impairment in patients with atrial fibrillation and diabetes mellitus [J]. The Journal of Practical Medicine, 2024, 40(16): 2236-2243.
[9]	Tingting YANG,Yidan HUANG,Rongrong YANG,Ying YANG,Jing. ZHANG. Study on the relationship between serum LRG1， LDH with periodontal indexes and periodontal lesions in patients with type 2 diabetes and chronic periodontitis [J]. The Journal of Practical Medicine, 2024, 40(16): 2250-2255.
[10]	Jing GUI,Feng WANG,Hui YANG,Yumao CAI,Chuangyue. HONG. Investigation of oxidised low⁃density lipoprotein as a risk assessment indicator in patients with type 2 diabetes mellitus combined with pulmonary tuberculosis [J]. The Journal of Practical Medicine, 2024, 40(14): 1995-2002.
[11]	Dechao LI,Mingjin ZHANG,Yibi LAN,Chunlei MA,Weijin. FU. Expression of allograft inflammatory factor⁃1 in the testicular model of diabetes mellitus rats [J]. The Journal of Practical Medicine, 2024, 40(1): 65-71.
[12]	WANG Lei , GUI Yaoyao, KOU Li. . Effect of exenatide combined with polyene phosphatidyl choline on serum HMGB1 and APN levels in pa⁃ tients with alcoholic fatty liver disease [J]. The Journal of Practical Medicine, 2023, 39(9): 1164-1168.
[13]	LIN Danhong, QUAN Huibiao, OU Qianying, CEN Chaoping, CHEN Kaining.. Effect of liraglutide on testosterone level and sexual function in overweight or obese men with type 2 diabe⁃ tes mellitus [J]. The Journal of Practical Medicine, 2023, 39(7): 894-898.
[14]	NIU Chunyan, CHEN Yao, SONG Yongqiang.. Key points of the American Association of Clinical Endocrinology Clinical Practice Guideline for the diagno⁃ sis and management of nonalcoholic fatty liver disease in primary care and endocrinology clinical settings co ⁃sponsored [J]. The Journal of Practical Medicine, 2023, 39(3): 267-273.
[15]	Yan YANG,Zhen WANG,Defeng. WANG. Clinical research progress of bexagliflozin： A new hypoglycemic drug [J]. The Journal of Practical Medicine, 2023, 39(22): 3015-3020.