Effect of 2-arachidonoylglycerol level on hippocampal neuroinflammatory response in autistic model rats

doi:10.11852/zgetbjzz2020-1372

Abstract

Abstract: Objective To analyze the effect of regulating 2-arachidonoylglycerol(2-AG) level on the expression of cytokines in hippocampus of autistic model rats,in order to provide a theoretical basis for the etiology research of autism spectrum disorders (ASD). Methods Autistic model rats induced by sodium valproate (VPA) were chosen as objects,and were divided into control group (CON group),VPA group,VPA + 1 mg/kg JZL184 group (VPA + 1RJ group),VPA + 3 mg/kg JZL184 group (VPA + 3RJ group),VPA + 10 mg/kg JZL184 group (VPA + 10RJ group). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to detect the level of 2-AG in the hippocampus. Real-time fluorescent quantitative PCR (RT-PCR) and enzyme-linked immuno sorbent assay (ELISA) were used to measure the mRNA expression levels and tissue contents in the hippocampus,including pro-inflammatory cytokines TNF-α,iNOS,IL-1β and anti-inflammatory cytokines Arg-1,IL-4,IL-10 and IL-13. Results LC-MS/MS results showed that the level of 2-AG decreased in hippocampus of VPA rats than that in the CON group (P<0.05),which was significantly higher in VPA+10RJ group than that in VPA group (P<0.05). RT-PCR results showed that compared with the CON group,the expression level of iNOS was increased in the VPA group (P<0.05). Compared with the VPA group,the expression levels of IL-1β,Arg-1,IL-10,IL-13 increased,while the expression level of iNOS decreased in the VPA + 1RJ group. In the VPA+3RJ group,TNF-α and iNOS expression levels decreased than those in VPA group. iNOS expression decreased,and IL-10 expression increased in the VPA + 10RJ group (P<0.05). ELISA results showed that compared with CON group,the expression of iNOS in VPA group was up-regulated,and the expression of Arg-1 and IL-10 was decreased (P<0.05). Compared with the VPA group,the expression of IL-4 in the VPA + 1RJ group was up-regulated,the expression of iNOS was reduced and the expression of IL-10 was up-regulated in the VPA + 3RJ group. In the VPA + 10RJ group,the expression of iNOS was reduced,and the expression of Arg-1 and IL-10 was up-regulated compared with VPA group (P<0.05). Conclusion Elevating 2-AG level could relieve the abnormal neuroinflammation in hippocampus of autistic model rats.

Key words: autism spectrum disorder, valiproic acid, neuroinflammation, 2-arachidonoylglycerol, cytokine

摘要： 目的分析花生四烯酸甘油酯(2-AG)水平升高对孤独症模型鼠海马神经炎性反应的影响,为孤独症谱系障碍(ASD)病因研究提供理论依据。方法以丙戊酸钠(VPA)诱导的孤独症模型鼠为研究对象,分为对照组(CON组)、VPA组、VPA+1 mg/kg JZL184组(VPA+1RJ组)、VPA+3 mg/kg JZL184组(VPA+3RJ组)、VPA+10 mg/kg JZL184组(VPA+10RJ组)。采用液相色谱-质谱联用技术(LC-MS/MS)检测海马组织中2-AG水平;实时荧光定量PCR(RT-PCR)和酶联免疫吸附实验(ELISA)检测海马组织中促炎因子TNF-α、iNOS、IL-1β和抗炎因子Arg-1、IL-4、IL-10和IL-13的mRNA表达水平和组织含量。结果 LC-MS/MS结果显示:VPA组海马组织中2-AG水平显著低于CON组(P＜0.05);VPA+10RJ组2-AG水平显著高于VPA组(P＜0.05)。RT-PCR结果显示:与CON组相比,VPA组中iNOS表达增加(P＜0.05)。与VPA组相比,VPA+1RJ组中IL-1β、Arg-1、IL-10、IL-13表达增加,iNOS表达降低(P<0.05);VPA+3RJ组中TNF-α和iNOS表达降低(P<0.05);VPA+10RJ组中iNOS表达降低,IL-10表达增加(P＜0.05)。ELISA结果显示:与CON组相比,VPA组中iNOS表达上调,Arg-1、IL-10表达降低(P＜0.05)。与VPA组相比,VPA+1RJ组中IL-4表达上调;VPA+3RJ组中iNOS表达降低,IL-10表达上调;VPA+10RJ组中iNOS表达降低,Arg-1、IL-10表达上调(P＜0.05)。结论 2-AG水平的升高可缓解孤独症模型鼠海马组织中存在的异常神经炎性反应。

关键词: 孤独症谱系障碍, 丙戊酸钠, 神经炎性反应, 花生四烯酸甘油酯, 细胞因子

CLC Number:

R285.5

LIU Yu, LIU Ze-hui, XIE Shu, LI Ling, SHI Mei, ZOU Ming-yang, SUN Cai-hong. Effect of 2-arachidonoylglycerol level on hippocampal neuroinflammatory response in autistic model rats[J]. Chinese Journal of Child Health Care, 2021, 29(4): 385-388.

刘瑜, 刘泽慧, 谢姝, 李玲, 施梅, 邹明扬, 孙彩虹. 花生四烯酸甘油酯水平对孤独症模型鼠海马神经炎性反应的影响[J]. 中国儿童保健杂志, 2021, 29(4): 385-388.

References

[1] Francesmonneris A,Pincus H,First M. Diagnostic and statistical manual of mental disorders:DSM-V[M]. Washington DC:American Psychiatric Association,2013.
[2] Maenner MJ,Shaw KA,Baio J,et al.Prevalence of autism spectrum disorder among children aged 8 years-autism and developmental disabilities monitoring network,11 sites,United States,2016[J].MMWR Surveill Summ,2020,69(4):1-12.
[3] Jacome MCI,Chacon LMM,Cuesta HV,et al.Peripheral inflammatory markers contributing to comorbidities in autism[J].Behav Sci (Basel),2016,6(4):29.
[4] Li X,Chauhan A,Sheikh AM,et al.Elevated immune response in the brain of autistic patients[J].J Neuroimmunol,2009,207(1-2):111-116.
[5] Nomura DK,Morrison BE,Blankman JL,et al.Endocannabinoid hydrolysis generates brain prostaglandins that promote neuroinflammation[J].Science,2011,334(6057):809-813.
[6] Rahmani MR,Shamsizadeh A,Moghadam-Ahmadi A,et al.JZL184,as a monoacylglycerol lipase inhibitor,down-regulates inflammation in a cannabinoid pathway dependent manner[J].Biomed Pharmacother,2018,103:1720-1726.
[7] 李德欣. eCB系统异常与ASD儿童认知及社交功能的相关性研究 [D]. 哈尔滨:哈尔滨医科大学,2020.
[8] 李玲. VPA诱导的孤独症模型鼠中内源性大麻素系统表达的差异研究[D]. 哈尔滨:哈尔滨医科大学,2020.
[9] Blankman JL,Simon GM,Cravatt BF. A comprehensive profile of brain enzymes that hydrolyze the endocannabinoid 2-arachidonoylglycerol[J].Chem Biol,2007,14(12):1347-1356.
[10] Hu CC,Xu X,Xiong GL,et al.Alterations in plasma cytokine levels in chinese children with autism spectrum disorder[J].Autism Res,2018,11(7):989-999.
[11] Xie J,Huang L,Li X,et al.Immunological cytokine profiling identifies TNF-alpha as a key molecule dysregulated in autistic children[J].Oncotarget,2017,8(47):82390-82398.
[12] Eissa N,Azimullah S,Jayaprakash P,et al.The dual-active histamine H3 receptor antagonist and acetylcholine esterase inhibitor E100 ameliorates stereotyped repetitive behavior and neuroinflammmation in sodium valproate induced autism in mice[J].Chem Biol Interact,2019,312:108775.
[13] Ahmad SF,Nadeem A,Ansari MA,et al.The potent immunomodulatory compound VGX-1027 regulates inflammatory mediators in CD4(+) T cells,which are concomitant with the prevention of neuroimmune dysregulation in BTBR T(+) Itpr3(tf)/J mice[J].Life Sci,2019,237:116930.
[14] Habib SS,Al-Regaiey K,Bashir S,et al.Role of endocannabinoids on neuroinflammation in autism spectrum disorder prevention[J].J Clin Diagn Res,2017,11(6):Ce1-ce3.
[15] Wilson RI,Nicoll RA. Endogenous cannabinoids mediate retrograde signalling at hippocampal synapses[J].Nature,2001,410(6828):588-592.
[16] Zamberletti E,Gabaglio M,Parolaro D. The endocannabinoid system and autism spectrum disorders:Insights from animal models[J].Int J Mol Sci,2017,18(9):1916.
[17] Mecha M,Feliú A,Machín I,et al.2-AG limits Theiler's virus induced acute neuroinflammation by modulating microglia and promoting MDSCs[J].Glia,2018,66(7):1447-1463.