摘要
在明确雷公藤红素(CEL)干预肥胖-抑郁共病小鼠杏仁核(AMY)及中缝背核(DRN)mRNA表达异同基础上,进一步揭示CEL干预肥胖-抑郁共病中枢炎症的药效靶标群。C57BL/6J小鼠随机分为正常(Chow)组,肥胖-抑郁共病(COM)组,CEL低、中、高剂量(CEL-L、CEL-M、CEL-H,0.5、1.0、2.0 mg·kg^(-1))组。Chow组给予普通饲料,其余组给予高脂饲料结合潮湿垫料慢性应激。饲养10周后灌胃给药3周,然后取Chow组、COM组和CEL-H组小鼠的AMY及DRN进行转录组分析,并将2个核团的目标差异基因以Venn图取交集。将交集基因导入STRING,进行蛋白-蛋白相互作用(PPI)分析,以DAVID进行基因本体论(GO)功能富集分析,明确CEL调控AMY、DRN的核心靶点。在独立样本中,通过qPCR对上述交集基因进行验证。结果显示,CEL调控AMY及DRN的共有基因为趋化因子家族Ccl2、Ccl5、Ccl7、Cxcl10、Cxcr6和Hsp70家族Hspa1a、Hspa1b及Myd88、Il2ra、Irf7、Slc17a8、Drd2、Parp9、Nampt。GO分析显示,前五位节点Ccl2、Cxcl10、Myd88、Ccl5、Irf7均参与免疫-炎症调控(P<0.01)。独立样品qPCR结果显示,在AMY中,与Chow组相比,COM组趋化因子家族、Hsp70、Myd88、Il2ra、Irf7、Slc17a8、Parp9、Nampt表达显著上调,Drd2有降低的趋势;与COM组相比,CEL-H组上述病理变化显著改善。在DRN中,与Chow组相比,COM组趋化因子家族、Hsp70、Myd88、Il2ra、Irf7、Parp9、Nampt表达显著下调,Slc17a8表达显著上调;与COM组相比,CEL-H组Cxcr6、Irf7、Drd2表达显著上调,Slc17a8表达显著下调。在AMY及DRN中,CEL对Irf7的表达抑制及激活均呈现剂量依赖性(R2分别为0.7098、0.9172)。上述研究结果显示,CEL可通过调控相同靶标蛋白的双向表达,干预肥胖-抑郁共病小鼠AMY的免疫激活及DRN的免疫抑制状态,从而有效改善神经炎症。
This study aims to further elucidate the efficacy targets of celastrol(CEL)intervention in central inflammation in mice with obesity-depression comorbiditiy,based on the differential mRNA expression in the amygdala(AMY)and dorsal raphe nucleus(DRN)after CEL intervention.C57BL/6J mice were randomly divided into a normal diet group(Chow),a obesity-depression comorbidity(COM)group,and low-,medium-,and high-dose CEL groups(CEL-L,CEL-M,CEL-H,0.5,1.0,2.0 mg·kg^(-1)).The Chow group received a normal diet,while the COM group and CEL-L,CEL-M,CEL-H groups received a high-fat diet combined with chronic stress from wet bedding.After 10 weeks of feeding,the mice were orally administered CEL for three weeks.Subsequently,the AMY and DRN of mice in the Chow,COM,and CEL-H groups were subjected to transcriptome analysis,and the intersection of target differentially expressed genes in both nuclei was visualized using a Venn diagram.The intersected genes were then imported into STRING for protein-protein interaction(PPI)analysis,and Gene Ontology(GO)analysis was performed using DAVID to identify the core targets regulated by CEL in the AMY and DRN.Independent samples were subjected to quantitative real-time PCR(qPCR)to validate the intersection genes.The results revealed that the common genes regulated by CEL in the AMY and DRN included chemokine family genes Ccl2,Ccl5,Ccl7,Cxcl10,Cxcr6,and Hsp70 family genes Hspa1a,Hspa1b,as well as Myd88,Il2ra,Irf7,Slc17a8,Drd2,Parp9,and Nampt.GO analysis showed that the top 5 nodes Ccl2,Cxcl10,Myd88,Ccl5,and Irf7 were all involved in immune-inflammation regulation(P<0.01).The qPCR results from independent samples showed that in the AMY,compared with the results in the Chow group,chemokine family genes,Hsp70,Myd88,Il2ra,Irf7,Slc17a8,Parp9,and Nampt were significantly up-regulated in the COM group,with Drd2 showing a decreasing trend;these pathological changes were significantly improved in the CEL-H group compared to the COM group.In the DRN,compared with the results in the Chow group,chemokine f
作者
张双盼
姚雪敏
朱春燕
林娜
ZHANG Shuang-pan;YAO Xue-min;ZHU Chun-yan;LIN Na(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;Institute of Chinese Materia Medica,China Academy of Chinese Medicine Sciences,Beijing 100700,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第14期3828-3836,共9页
China Journal of Chinese Materia Medica
基金
北京市自然科学基金项目(7212185)。
