间歇禁食法在改善奥氮平诱导小鼠代谢紊乱中的机制研究

doi:10.12300/j.issn.1674-5817.2022.089

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (1): 3-10.DOI: 10.12300/j.issn.1674-5817.2022.089

• 人类疾病动物模型：药学专题 • 上一篇下一篇

间歇禁食法在改善奥氮平诱导小鼠代谢紊乱中的机制研究

李晗¹, 张笑瑞², 张成芳³()()

^1.上海交通大学医学院附属精神卫生中心, 上海市重性精神病重点实验室, 上海 201108
^2.中国科学院分子细胞科学卓越创新中心动物实验技术平台, 上海 200031
^3.同济大学附属精神卫生中心, 上海市浦东新区精神卫生中心, 同济大学精神疾病临床研究中心, 上海 200124

收稿日期:2022-06-20 修回日期:2022-07-21 出版日期:2023-02-25 发布日期:2023-03-09
通讯作者: 张成芳（1987—），女，博士研究生，主治医师，研究方向：抗精神病药物代谢不良反应的机制和临床防治。E-mail：callup1987@126.com。ORCID： 0000-0002-5977-0312
作者简介:李晗（1984—），男，硕士，助理研究员，研究方向：精神疾病模型研究。E-mail：erjunda140@126.com。ORCID: 0000-0003-3903-5254
基金资助:
上海市浦东新区优秀青年医学人才培养计划(PWRq2020-19);上海市浦东新区科技发展基金（民生科研专项：医药卫生）“间歇限制热量法在奥氮平诱导的胰岛素抵抗小鼠模型中的作用”(PKJ2018-Y27)

Mechanism of Intermittent Fasting in Improving Olanzapine-induced Metabolic Disorders in Mice

Han LI¹, Xiaorui ZHANG², Chengfang ZHANG³()()

^1.Shanghai Mental Health Center, Shanghai Jiao Tong University of Medicine, Shanghai Key Laboratory of Psychotic Disorders, Shanghai 201108, China
^2.Animal Core Facility of Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China
^3.Clinical Research Center for Mental Disorders, Shanghai Pudong New Area Mental Health Center, School of Medicine, Tongji University, Shanghai 200124, China

Received:2022-06-20 Revised:2022-07-21 Published:2023-02-25 Online:2023-03-09
Contact: ZHANG Chengfang (ORCID: 0000-0002-5977-0312), E-mail: callup1987@126.com

摘要/Abstract

摘要：

目的探究间歇禁食（intermittent fasting，IF）对奥氮平诱导小鼠代谢紊乱的保护作用及其机制。方法将C57BL/6J小鼠随机分为4组：生理盐水+自由摄食组（Saline+ Ad libitum）、生理盐水+间歇禁食组（Saline+IF）、奥氮平给药+自由摄食组（Olanzapine+ Ad libitum）、奥氮平给药+间歇禁食组（Olanzapine+IF），每组8只。其中间歇禁食组采用5∶2方案，即每周的周一、周四禁食，其余时间自由摄食，干预12周；以自由摄食作为间歇禁食的对照（Control），以生理盐水灌胃作为奥氮平给药的对照（Saline）。比较奥氮平处理组与对照组经过间歇禁食干预后小鼠体质量、肝脏质量和附睾旁脂肪组织质量的差异，并分别采用磁共振和HE染色法分析小鼠体脂、瘦素以及内脏脂肪浸润的变化；同时，分别采用葡萄糖氧化酶法和放射免疫法测定小鼠糖代谢过程中空腹血糖、胰岛素和胰岛素抵抗指数（HOMA-IR）的水平差异，分别采用ELISA和实时荧光定量PCR法测定H₂O₂含量和线粒体损伤相关标志物细胞色素C（Cytochrome C）mRNA水平。结果分组处理12周后，奥氮平诱导小鼠体质量、体脂、瘦素和内脏脂肪浸润明显增加（P＜0.05），空腹血糖、胰岛素和胰岛素指数也明显增加（P＜0.05）；然而间歇禁食可以使上述指标明显降低（P＜0.05）。进一步研究表明：间歇禁食处理后小鼠脂肪组织部位的H₂O₂释放和Cytochrome C mRNA表达水平均明显降低（P＜0.05）。结论间歇禁食可以减轻奥氮平引起的小鼠代谢紊乱，其机制可能与抑制氧化应激水平和维持线粒体功能有关。

关键词: 间歇禁食法, 肥胖, 氧化应激, 线粒体, C57BL/6J小鼠

Abstract:

Objective To explore the beneficial role and potential mechanism of intermittent fasting in olanzapine-induced metabolic disorders. Methods C57BL/6J mice were randomly divided into four groups: Saline + ad libitum (Saline+Ad libitum), Saline + intermittent fasting (Saline +IF), olanzapine administration + ad libitum (Olanzapine+ Ad libitum), and olanzapine administration + intermittent fasting (Olanzapine+IF)， with eight mice in each group. The IF group adopted the 5∶2 scheme, that is, fasting on Monday and Thursday every week, and eating freely in the rest of the time. Ad libitum feeding as the control of intermittent fasting, Saline gavage as the control of olanzapine administration. The experiment lasted for 12 weeks. The differences of body mass, liver mass and epididymal adipose tissue mass were compared between the olanzapine-treated group and the control group after IF intervention. The body fat mass, lean body mass, and visceral fat infiltration of mice were analyzed by nuclear magnetic resonance and HE staining, respectively. Furthermore, the levels of fasting blood glucose, insulin, and insulin resistance index (HOMA-IR) in the process of glucose metabolism were also measured by glucose oxidase method and radioimmunoassay, respectively. The effects of IF on H₂O₂ release and the level of cytochrome C mRNA, a marker related to mitochondrial damage, were detected by ELISA and real-time fluorescence quantitative PCR. Results After 12 weeks of treatment, olanzapine induced a significant increase in body mass, body fat, lean body mass and visceral fat infiltration (P<0.05), as well as fasting blood glucose, insulin, and HOMA-IR (P<0.05); however, IF significantly reduced the above indicators (P<0.05). Further studies showed that the release of H₂O₂ and the expression of Cytochrome C mRNA in adipose tissue of mice after intermittent fasting treatment were significantly decreased (P<0.05). Conclusion Intermittent fasting therapy can alleviate olanzapine-induced metabolic disorders in mice. The underlying mechanism may involve the inhibition of oxidative stress level and the maintenance of mitochondrial functions.

Key words: Intermittent fasting therapy, Obesity, Oxidative stress, Mitochondria, C57BL/6J mice

中图分类号:

Q95-33

李晗, 张笑瑞, 张成芳. 间歇禁食法在改善奥氮平诱导小鼠代谢紊乱中的机制研究[J]. 实验动物与比较医学, 2023, 43(1): 3-10.

Han LI, Xiaorui ZHANG, Chengfang ZHANG. Mechanism of Intermittent Fasting in Improving Olanzapine-induced Metabolic Disorders in Mice[J]. Laboratory Animal and Comparative Medicine, 2023, 43(1): 3-10.

图/表 4

图1 间歇禁食对奥氮平小鼠体质量、体脂分布和内脏脂肪浸润的影响注：A，连续12周记录不同处理组小鼠的体质量变化（*F=161.20，P<0.001）；B，奥氮平摄入12周后间歇禁食对小鼠体质量的影响（*F=62.87，P<0.001）；C，奥氮平摄入12周后小鼠脂肪含量的变化（*F=0.027，P<0.001）；D，奥氮平摄入12周后小鼠瘦素含量（nsF=78.54，P=0.994）；E，奥氮平摄入12周后小鼠肝脏和附睾旁脂肪细胞形态（HE染色，比例尺大小均为200 μm）。

Figure 1 Effects of intermittent fasting on body mass， body fat distribution， visceral fat infiltration in olanzapine-induced miceNote：A， Body mass changes of mice in different treatment groups for 12 consecutive weeks （F=161.20， *P<0.001， two-way ANOVA followed by Turkey's post-test）. B， Effect of intermittent fasting on body mass of mice with olanzapine diet for 12 weeks （*F=62.87，P<0.001， one-way ANOVA followed by Turkey's post-test）. C， The changes of fat mass in mice after olanzapine diet for 12 weeks （*F=0.027，P<0.001， one-way ANOVA followed by Turkey's post-test）. D， The changes of lean mass in mice after olanzapine diet for 12 weeks （n.sF=78.54， P=0.994，one-way ANOVA followed by Turkey's post-test）. E， Liver and epididymal adipose morphology after olanzapine diet for 12 weeks（HE staining，the scale bar is 200 μm）.

图2 间歇禁食对奥氮平小鼠空腹血糖、胰岛素和胰岛素抵抗水平的影响注：间歇禁食干预12周后，检测小鼠空腹胰岛素含量（A）（*F=18.08，P<0.001）、胰岛素抵抗的稳态模型评估（HOMA-IR）指数［空腹血糖（mmol/L）×空腹胰岛素（mU/L）/22.5］（B）（*F=9.707，P=0.000 4），以及小鼠血糖含量（C）（n.sF=0.860，P=0.477）.

Figure 2 Effects of intermittent fasting on fasting blood glucose, insulin, and HOMA-IR, in olanzapine-induced miceNote：After 12 weeks of intermittent fasting， detecting fasting insulin levels （A）（*F=18.08，P<0.001， one-way ANOVA followed by Turkey's post-test）， homeostatic model assessment of insulin resistance （HOMA-IR）（B）（*F=9.707， P=0.000 4， one-way ANOVA followed by Turkey's post-test）， and fasting glucose levels （C）（n.sF=0.860， P=0.477， one-way ANOVA followed by Turkey's post-test）.

图3 间歇性禁食对奥氮平小鼠氧化应激因子和线粒体功能的影响注：间歇禁食12周后，ELISA检测小鼠H2O2释放量（A）（*F=4.633，P=0.016），实时定量PCR检测Cytochrome C的mRNA转录水平（B）（*F=3.605，P=0.031）。

Figure 3 Effects of intermittent fasting on oxidative stress， mitochondrial function in olanzapine-induced miceNote：Hydrogen peroxide release was detected by ELISA （A）（*F=4.633， P=0.016）， and the transcriptional level of Cytochrome C mRNA was detected by real-time fluorescence quantitative PCR （B）（*F=3.605， P=0.031） after 12 weeks of intermittent fasting treatment in mice.

图4 间歇禁食法改善奥氮平诱导小鼠代谢紊乱的潜在通路模式图

Figure 4 Schematic illustration of potential pathways underlying the mechanism of intermittent fasting in ameliorating olanzapine-induced metabolic disorders in mice

参考文献 20

1	STĘPNICKI P, KONDEJ M, KACZOR A A. Current concepts and treatments of schizophrenia[J]. Molecules, 2018, 23(8): E2087. DOI:10.3390/molecules23082087 .
2	JOHNSEN E, KROKEN R A, LØBERG E M, et al. Amisulpride, aripiprazole, and olanzapine in patients with schizophrenia- spectrum disorders (BeSt InTro): a pragmatic, rater-blind, semi-randomised trial[J]. Lancet Psychiatry, 2020, 7(11):945- 954. DOI:10.1016/S2215-0366(20)30341-2 .
3	CHENG Z, YUAN Y B, HAN X, et al. An open-label randomised comparison of aripiprazole, olanzapine and risperidone for the acute treatment of first-episode schizophrenia: eight- week outcomes[J]. J Psychopharmacol, 2019, 33(10):1227- 1236. DOI:10.1177/0269881119872193 .
4	POZNYAK A, GRECHKO A V, POGGIO P, et al. The diabetes mellitus-atherosclerosis connection: the role of lipid and lucose metabolism and chronic inflammation[J]. Int J Mol Sci, 2020, 21(5):1835. DOI:10.3390/ijms21051835 .
5	STANCU I C, CREMERS N, VANRUSSELT H, et al. Aggregated Tau activates NLRP3-ASC inflammasome exacerbating exogenously seeded and non-exogenously seeded Tau pathology in vivo[J]. Acta Neuropathol, 2019, 137(4):599-617. DOI:10.1007/s00401-018-01957-y .
6	BARRA N G, HENRIKSBO B D, ANHÊ F F, et al. The NLRP3 inflammasome regulates adipose tissue metabolism[J]. Bio chem J, 2020, 477(6):1089-1107. DOI:10.1042/BCJ20190472 .
7	MEYERS A K, ZHU X W. The NLRP3 inflammasome: metabolic regulation and contribution to inflammaging[J]. Cells, 2020, 9 (8):1808. DOI:10.3390/cells9081808 .
8	LIU Q Y, ZHANG D Y, HU D Y, et al. The role of mitochondria in NLRP3 inflammasome activation[J]. Mol Immunol, 2018, 103:115-124. DOI:10.1016/j.molimm.2018.09.010 .
9	MILLS E L, KELLY B, LOGAN A, et al. Succinate dehydrogenase supports metabolic repurposing of mitochondria to drive inflammatory macrophages[J]. Cell, 2016, 167(2):457-470.e13. DOI:10.1016/j.cell.2016.08.064 .
10	ANTONI R, JOHNSTON K L, COLLINS A L, et al. Effects of intermittent fasting on glucose and lipid metabolism[J]. Proc Nutr Soc, 2017, 76(3):361-368. DOI:10.1017/S002966511 6002986 .
11	LIANG B J, LIAO S R, HUANG W X, et al. Intermittent fasting therapy promotes insulin sensitivity by inhibiting NLRP3 in flammasome in rat model[J]. Ann Palliat Med, 2021, 10(5): 5299-5309. DOI:10.21037/apm-20-2410 .
12	HUANG Y Q, WANG Y, WANG H, et al. Prevalence of mental disorders in China: a cross-sectional epidemiological study [J]. Lancet Psychiatry, 2019, 6(3):211-224. DOI:10.1016/S2215- 0366(18)30511-X .
13	徐若愚, 李献云. 精神分裂症认知行为治疗的研究进展[J]. 神经疾病与精神卫生, 2022, 22(5):342-346.
	XU R Y, LI X Y. Research progress of cognitive behavioral therapy for schizophrenic[J]. J Neurosci Ment Health, 2022, 22 (5):342-346.
14	CARTERI R B, MENEGASSI L N, FELDMANN M, et al. Inter mittent fasting promotes anxiolytic-like effects unrelated to synaptic mitochondrial function and BDNF support[J]. Behav Brain Res, 2021, 404:113163. DOI:10.1016/j.bbr.2021.113163 .
15	MATTSON M P, LONGO V D, HARVIE M. Impact of intermit tent fasting on health and disease processes[J]. Ageing Res Rev, 2017, 39:46-58. DOI:10.1016/j.arr.2016.10.005 .
16	WU K K L, CHEUNG S W M, CHENG K K Y. NLRP3 inflammasome activation in adipose tissues and its implications on meta bolic diseases[J]. Int J Mol Sci, 2020, 21(11):4184. DOI:10.3390/ ijms21114184 .
17	ROTHMAN S M, GRIFFIOEN K J, WAN R Q, et al.
	Brainderived neurotrophic factor as a regulator of systemic
	and brain energy metabolism and cardiovascular health[J].
	N Y Acad Sci Ann, 2012, 1264(1):49-63. DOI:10.1111/j.1749-6632 .
	2012.06525.x.
18	LONGO V D, MATTSON M P. Fasting: molecular mechanisms and clinical applications[J]. Cell Metab, 2014, 19(2):181-192. DOI:10.1016/j.cmet.2013.12.008 .
19	ZHANG W J, CHEN S J, ZHOU S C, et al. Inflammasomes and fibrosis[J]. Front Immunol, 2021, 12:643149. DOI:10.3389/ fimmu.2021.643149 .
20	KAWAI T, AUTIERI M V, SCALIA R. Adipose tissue inflammation and metabolic dysfunction in obesity[J]. Am J Physiol Cell Physiol, 2021, 320(3): C375-C391. DOI:10.1152/ajpcell.00379.2020 .

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间歇禁食法在改善奥氮平诱导小鼠代谢紊乱中的机制研究

Mechanism of Intermittent Fasting in Improving Olanzapine-induced Metabolic Disorders in Mice

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