左归降糖解郁方调控CD300f/GLUT1信号通路改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制

刘检; 唐林; 赵洪庆; 姜帆; 刘林; 胡超

doi:10.3969/j.issn.1006-2157.2024.11.012

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左归降糖解郁方调控CD300f/GLUT1信号通路改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制

实验研究 | 更新时间：2024-12-02

- 左归降糖解郁方调控CD300f/GLUT1信号通路改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制
- Zuogui Jiangtang Jieyu Formula regulates the CD300f/GLUT1 signaling pathway to improve the synaptic damage of hippocampal neurons in rats with diabetes-related depression
- 北京中医药大学学报 2024年47卷第11期页码：1573-1584
- 作者机构：
  
  1.湖南中医药大学第一附属医院，长沙　410007
  2.湖南中医药大学科技创新中心
- 作者简介：
  
  刘检，男，副研究员，博士，硕士生导师
  #胡超，女，硕士，主管药师，主要研究方向：中药防治慢性重大疾病，E-mail: 348009051@qq.com
- 基金信息：
  
  国家自然科学基金青年项目(82104793);湖南省自然科学基金面上项目(2022JJ30451);湖南省三尖创新人才工程湖湘青年英才项目(2022RC1226)
- DOI：10.3969/j.issn.1006-2157.2024.11.012
  中图分类号： R285.5
- 收稿日期：2024-05-31，
  
  纸质出版日期：2024-11-30
- 稿件说明：
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刘检, 唐林, 赵洪庆, 等. 左归降糖解郁方调控CD300f/GLUT1信号通路改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制[J]. 北京中医药大学学报, 2024,47(11):1573-1584.

LIU Jian, TANG Lin, ZHAO Hongqing, et al. Zuogui Jiangtang Jieyu Formula regulates the CD300f/GLUT1 signaling pathway to improve the synaptic damage of hippocampal neurons in rats with diabetes-related depression[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(11): 1573-1584.
刘检, 唐林, 赵洪庆, 等. 左归降糖解郁方调控CD300f/GLUT1信号通路改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制[J]. 北京中医药大学学报, 2024,47(11):1573-1584. DOI： 10.3969/j.issn.1006-2157.2024.11.012.

LIU Jian, TANG Lin, ZHAO Hongqing, et al. Zuogui Jiangtang Jieyu Formula regulates the CD300f/GLUT1 signaling pathway to improve the synaptic damage of hippocampal neurons in rats with diabetes-related depression[J]. Journal of beijing university of traditional chinese medicine, 2024, 47(11): 1573-1584. DOI： 10.3969/j.issn.1006-2157.2024.11.012.

摘要

目的

基于白细胞单免疫球蛋白样受体3(CD300f)/葡萄糖转运体1(GLUT1)信号通路介导小胶质细胞糖代谢，探讨左归降糖解郁方改善糖尿病并发抑郁症大鼠海马神经元突触损伤的作用机制。

方法

80只雄性SD大鼠按照随机数字表法选取10只大鼠作为正常组，其余70只高脂饲料喂养4周后单次尾静脉注射链脲佐菌素(38 mg/kg)复制大鼠糖尿病模型，筛选造模成功的大鼠60只，按随机数字表法分为模型组、CD300f阻断剂组、CD300f激动剂组、二甲双胍＋氟西汀组(二甲双胍0.18 g/kg＋氟西汀1.8 mg/kg)和左归降糖解郁方高、低剂量组(20.52、10.26 g/kg)。除正常组外，其余各组大鼠予28 d慢性不可预知温和应激加孤养复制糖尿病并发抑郁症大鼠模型。其中，二甲双胍＋氟西汀组与左归降糖解郁方高、低剂量组在造模第2周后连续灌胃相应药物，正常组、模型组灌胃等体积蒸馏水，连续14 d；CD300f阻断剂组、激动剂组则进行海马区微量注射给药，分别注射髓系细胞触发受体抑制因子(CLM1，2 μg/kg)和免疫球蛋白Fc段表面蛋白(Fcγ，5 μg/kg)，每周1次。干预结束后，采用旷场实验、强迫游泳实验检测大鼠抑郁样行为；生化分析检测大鼠海马组织葡萄糖、乳酸含量及腺苷二磷酸(ADP)/三磷酸腺苷(ATP)；采用酶联免疫吸附测定法检测大鼠海马组织胰岛素、5－羟色胺(5-HT)和多巴胺(DA)水平；采用免疫荧光检测海马组织CD300f、GLUT1、突触相关前膜蛋白3(RIMS3)、突触后膜相关蛋白102(SAP102)平均荧光强度；蛋白质印迹法检测大鼠海马组织CD300f、GLUT1、RIMS3、SAP102蛋白表达情况；采用尼氏染色和透射电镜观察大鼠海马组织病理改变。

结果

与正常组比较，模型组大鼠旷场总活动路程减少、强迫游泳不动时间增加，海马组织葡萄糖、乳酸含量及ADP/ATP升高、胰岛素水平降低、5-HT和DA水平下降，海马组织CD300f、GLUT1、RIMS3、SAP102平均荧光强度及蛋白相对表达量均降低(均

0.05)，海马神经元突触超微结构受损。与模型组比较，CD300f激动剂组及左归降糖解郁方高、低剂量组上述抑郁样行为学改变、糖代谢和单胺神经递质失衡得到改善(均

0.05)；CD300f激动剂组及左归降糖解郁方高剂量组海马CD300f、GLUT1、RIMS3、SAP102平均荧光强度和蛋白相对表达量均升高(均

0.05)，突触损伤减轻；CD300f阻断剂组葡萄糖、乳酸、ADP/ATP、5-HT异常及CD300f蛋白表达异常情况加重(均

0.05)，突触损伤加重。

结论

左归降糖解郁方能有效缓解糖尿病并发抑郁症大鼠海马小胶质细胞糖代谢异常与神经元突触损伤，其机制可能与调控CD300f/GLUT1信号通路有关。

Abstract

Objective

To explore the protective mechanism of

Zuogui Jiangtang Jieyu

Formula (ZGF) on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3 (CD300f)/glucose transporter 1 (GLUT1) signal-mediated microglial glucose metabolism in rats with diabetes-related depression.

Methods

Eighty male SD rats were randomly selected using random number table method

with 10 rats serving as the normal group. The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model. Sixty rats were screened and successfully modeled

which were randomly divided into the model

CD300f blocker

CD300f agonist

metformin + fluoxetine (metformin 0.18 g/kg + fluoxetine 1.8 mg/kg)

and ZGF high- and low-dose (20.52 and 10.26 g/kg

respectively) groups using random number table method. In addition to the normal group

the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model. The metformin + fluoxetine and ZGF high- and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling. The normal and model groups were administered an equal amount of distilled water by gavage. The CD300f blocker group and agonist group received microinjection into the hippocampus

with injection of myeloid cell trigger receptor inhibitory factor (CLM1

2 μg/kg) and immunoglobulin Fc surface protein (Fcγ

5 μg/kg) once a week

respectively. Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention. Biochemical analysis was used to detect the glucose

lactic acid

and adenosine diphosphate (ADP)/adenosine triphosphate (ATP) ratio contents. The insulin

5-hydroxytryptamine (5-HT)

and dopamine (DA) levels in the hippocampus were detected using an enzyme-linked immunosorbent assay. Immunofluorescence was used to detect the average fluorescence intensity of CD300f

GLUT1

regulating synaptic membrane wxocytosis 3 (RIMS3)

and synapse-associated protein 102 (SAP102) in hippocampal tissue. Western blotting was used to detect the CD300f

GLUT1

RIMS3

and SAP102 protein expression levels in the hippocampus. The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.

Results

Compared with the normal group

the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time

with an increase in glucose and lactic acid contents and ADP/ATP ratio

whereas a decrease in insulin

5-HT

and DA levels was observed in the hippocampus. The average fluorescence intensity and relative protein expression levels of CD300f

GLUT1

RIMS3

and SAP102 in hippocampal tissue decreased (

0.05)

and the synaptic ultrastructure of hippocampal neurons was damaged. Compared with the model group

depression-like behavioral changes

glucose metabolism

and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high- and low-dose group (

0.05). The average fluorescence intensity and relative protein expression levels of CD300f

GLUT1

RIMS3

and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased (

0.05)

and synaptic damage was alleviated. The abnormal levels of glucose

lactate

ADP/ATP

5-HT

and CD300f protein expression were aggravated in the CD300f blocker group (

0.05)

and synaptic damage was aggravated.

Conclusion

ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression. Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.

关键词

Keywords

references

BELKO S , HUTCHINSON M , HAYDEN G , et al . Co-designing diabetes care with patients [J ] . J Diabetes Sci Technol , 2024 , 18 ( 1 ): 53 － 58 .

XU HJ , PAN JH , CHEN Q . The progress of clinical research on the detection of 1, 5－anhydroglucitol in diabetes and its complications [J ] . Front Endocrinol , 2024 , 15 : 1383483 .

ALI MK , PEARSON－STUTTARD J , SELVIN E , et al . Interpreting global trends in type 2 diabetes complications and mortality [J ] . Diabetologia , 2022 , 65 ( 1 ): 3 － 13 .

PRABU P , POONGOTHAI S , SHANTHIRANI CS , et al . Altered circulatory levels of miR－128, BDNF, cortisol and shortened telomeres in patients with type 2 diabetes and depression [J ] . Acta Diabetol , 2020 , 57 ( 7 ): 799 － 807 .

BAI S , WANG JS , LIU JT , et al . Analysis of depression incidence and influence factors among middle-aged and elderly diabetic patients in China: based on CHARLS data [J ] . BMC Psychiatry , 2024 , 24 ( 1 ): 146 .

VAN DER FELTZ－CORNELIS C , ALLEN SF , HOLT RIG , et al . Treatment for comorbid depressive disorder or subthreshold depression in diabetes mellitus: systematic review and meta-analysis [J ] . Brain Behav , 2021 , 11 ( 2 ): e01981 .

MOSILI P , MKHIZE BC , SIBIYA NH , et al . Review of the direct and indirect effects of hyperglycemia on the HPA axis in T2DM and the co-occurrence of depression [J ] . BMJ Open Diabetes Res Care , 2024 , 12 ( 1 ): e003218 .

LI SX , YANG D , ZHOU XH , et al . Neurological and metabolic related pathophysiologies and treatment of comorbid diabetes with depression [J ] . CNS Neurosci Ther , 2024 , 30 ( 4 ): e14497 .

WANG J , ZHOU DP , DAI ZJ , et al . Association between systemic immune-inflammation index and diabetic depression [J ] . Clin Interv Aging , 2021 , 16 : 97 － 105 .

SU WJ , LI JM , ZHANG T , et al . Microglial NLRP3 inflammasome activation mediates diabetes-induced depression-like behavior via triggering neuroinflammation [J ] . Prog Neuropsychopharmacol Biol Psychiatry , 2023 , 126 : 110796 .

蔺晓源，韩远山，孟盼，等 . 糖尿病并发抑郁症“虚、瘀、郁”的中医病机探讨 [J ] . 时珍国医国药， 2016 , 27 ( 8 ): 1942 － 1943 .

李萍，刘洋，邹蔓姝，等 . 基于CX3CL1－CX3CR1轴探讨左归降糖解郁方对糖尿病并发抑郁症模型大鼠的神经保护作用及机制 [J ] . 中国中药杂志， 2023 , 48 ( 21 ): 5822 － 5829 .

刘洋，李萍，杨蕙，等 . 左归降糖解郁方改善糖尿病并发抑郁症大鼠N－甲基D－天冬氨酸受体过激致海马突触损伤的作用机制 [J ] . 中草药， 2023 , 54 ( 19 ): 6323 － 6335 .

杨蕙，王宇红，杜青，等 . 左归降糖解郁方通过调控SIRT1/TORC1通路增加糖尿病并发抑郁症大鼠海马神经元树突棘密度 [J ] . 中国病理生理杂志， 2021 , 37 ( 3 ): 400 － 407 .

刘检，张尚霞，刘林，等 . 基于线粒体自噬介导的凋亡探讨左归降糖解郁方对糖尿病并发抑郁证海马突触微环境损伤的保护作用 [J ] . 中华中医药杂志， 2021 , 36 ( 4 ): 2012 － 2018 .

刘检，王宇红，韩远山，等 . 左归降糖解郁方对糖尿病并发抑郁症海马神经元CaMKⅡ及SynGap的调控作用 [J ] . 北京中医药大学学报， 2017 , 40 ( 5 ): 392 － 398 .

PAIVA IHR , MACIEL LM , SILVA RSD , et al . Prebiotics modulate the microbiota-gut-brain axis and ameliorate anxiety and depression-like behavior in HFD-fed mice [J ] . Food Res Int , 2024 , 182 : 114153 .

YANG Z , SHENG JH , IET FELLOW , et al . Glucose-oxygen coupling can serve as a biomarker for neuroinflammation-related genetic variants [J ] . Cerebral Cortex , 2024 ， 34 ( 2 )： bhad520 .

ZSOMBOK A , DESMOULINS LD , DERBENEV AV . Sympathetic circuits regulating hepatic glucose metabolism: where we stand [J ] . Physiol Rev , 2024 , 104 ( 1 ): 85 － 101 .

RAJKHOWA B , MEHAN S , SETHI P , et al . Activation of SIRT－1 signalling in the prevention of bipolar disorder and related neurocomplications: target activators and influences on neurological dysfunctions [J ] . Neurotox Res , 2022 , 40 ( 2 ): 670 － 686 .

DEMPSEY LA . CD300f in depression [J ] . Nat Immunol , 2020 , 21 ( 5 ): 488 .

LAGO N , KAUFMANN FN , NEGRO-DEMONTEL ML , et al . CD300f immunoreceptor is associated with major depressive disorder and decreased microglial metabolic fitness [J ] . Proc Natl Acad Sci U S A , 2020 , 117 ( 12 ): 6651 － 6662 .

吴丹，高耀，邢婕，等 . 逍遥散治疗肝郁脾虚型抑郁症的药理作用机制研究进展 [J ] . 中国实验方剂学杂志， 2019 , 25 ( 8 ): 187 － 193 .

柳辰玥 . 基于Homer1－mGluR5及mTOR通路探讨逍遥散对抑郁症肝郁脾虚证大鼠神经保护作用机理 [D ] . 北京：北京中医药大学， 2021 .

LYRA E SILVA NM , LAM MP , SOARES CN , et al . Insulin resistance as a shared pathogenic mechanism between depression and type 2 diabetes [J ] . Front Psychiatry , 2019 , 10 : 57 .

LEE EJ , PARK JS , LEE YY , et al . Anti-inflammatory and anti-oxidant mechanisms of an MMP-8 inhibitor in lipoteichoic acid-stimulated rat primary astrocytes: involvement of NF-κB, Nrf2, and PPAR－γ signaling pathways [J ] . J Neuroinflammation , 2018 , 15 ( 1 ): 326 .

TODOROVA V , BLOKLAND A . Mitochondria and synaptic plasticity in the mature and aging nervous system [J ] . Curr Neuropharmacol , 2017 , 15 ( 1 ): 166 － 173 .

EVANS F , ALÍ－RUIZ D , REGO N , et al . CD300f immune receptor contributes to healthy aging by regulating inflammaging, metabolism, and cognitive decline [J ] . Cell Rep , 2023 , 42 ( 10 ): 113269 .

CIZERON M , QIU Z , KONIARIS B , et al . A brainwide atlas of synapses across the mouse life span [J ] . Science , 2020 , 369 ( 6501 ): 270 － 275 .

MICHEVA KD , WEINBERG RJ , SMITH SJ . A synapse census for the ages [J ] . Science , 2020 , 369 ( 6501 ): 253 － 254 .

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