1.Improvement of depressive-like behavior in Hemerocallis citrina Baroni using a zebrafish model
Hanwen ZHENG ; Zihang WANG ; Xinyue LIU ; Jiawei YU ; ALBERTO Carlos Pires Dias ; Fengzhong WANG ; Qiong WANG
Chinese Journal of Comparative Medicine 2024;34(4):20-27
Objective To investigate the effects of alcoholic extract of Hemerocallis citrina Baroni on the depressive-like behaviors in zebrafish larvae(Danio rerio)induced by reserpine.Methods Zebrafish larvae were divided into various groups:control(Con)group,reserpine group,fluoxetine group,H.citrina alcohol extract(HCE)low dose group(1.5 mg/L),HCE medium dose group(3 mg/L),and HCE high dose group(4.5 mg/L).Depressive-like behaviors were analyzed using sound and light stimulation.Real-time PCR was used to investigate the effects of HCE on depression related astrocyte markers(GFAP,C3,C4B,EMP-1,S100α-10)and the neurotrophic factor BDNF and its receptor genes(P75,TrkB).Results In comparison to the control group,the model group demonstrated significantly shorter movement distance and reduced movement time under sound and light stimulation(P<0.05,P<0.001,P<0.0001).Following the administration of HCE,zebrafish larvae exhibited significantly heightened sensitivity to light and sound stimulation compared to the model group(P<0.05,P<0.0001).Astrocyte marker genes were up-regulated in the model group zebrafish brains compared to the control group(P<0.0001).However,when the model group was administered HCE,the expression of astrocyte markers was significantly down-regulated compared to the model group(P<0.0001).Neurotrophic factor and its receptor genes(BDNF,P75,TrkB)were down-regulated in zebrafish brains in the model group compared to those in the control group(P<0.0001).However,in the group administered HCE,the expression of BDNF,P75,and TrkB was significantly up-regulated compared to that in the model group(P<0.01,P<0.0001).These findings suggest that HCE suppressed the inflammatory responses caused by astrocyte activation and promoted the production of neurotrophic factors and their receptor genes,thereby exerting an ameliorative effect on depression.Conclusions Alcoholic extracts of H.citrina can ameliorate the depression-like behavioral changes induced by reserpine in zebrafish larvae.They reduce the expression of astrocyte markers in the zebrafish brain and promote the production of neurotrophic factors and their receptor genes,playing an antidepressant role.
2.Exploring the mechanism of action of sour jujube nut compound formula for depression based on network pharmacology and experimental validation
Hanwen ZHENG ; Xinyue LIU ; Haiyan ZHAO ; Jiayin WANG ; Fulong LUO ; Bei FAN ; Alberto Carlos Pires Dias ; Fengzhong WANG ; Qiong WANG
Acta Laboratorium Animalis Scientia Sinica 2024;32(7):901-912
Objective In this study,we aimed to use network pharmacology techniques to predict the key targets of a prescription of Ziziphi spinosae semen formula(ZSSF)compound for depression,and to verify its mechanism of action using a zebrafish model of rifampicin-induced depression.Methods The drug targets of ZSSF were retrieved from the TCMSP database,and the target names were corrected using the UniProt database.Depression-related targets were identified using the GeneCards,OMIM,and NCBI databases.Protein-protein interaction information for the shared targets was predicted using the STRING database.The collected data were then analyzed using the Metascape database to determine GO and KEGG pathway enrichment,and the result were visualized using microbiotics.Behavioral experiments and reverse-transcription quantitative PCR experiments were conducted to verify the therapeutic effects of ZSSF on a zebrafish depression model induced by risperdal.Results 188 targets were screened to find the interactions between depression and ZSSF.The protein-protein interaction result showed that ZSSF primarily targeted TNF-α,IL-2,IL-6,IL-1β,and IL-10 to produce its antidepressant effect.KEGG pathway enrichment analysis revealed that ZSSF exerted its effects on depression through various signaling pathways,including the TNF,PI3K-Akt,and cGMP-PKG signaling pathways.The result of the animal experiments showed that the treatment groups given high,medium,and low doses of ZSSF exhibited significant improvements in movement distance under acoustic and light stimulation compared with the model group(P<0.05).The speed of movement of the treatment groups was also significantly faster(P<0.01).Additionally,the mRNA expression levels of TNF-α,IL-2,IL-6,IL-1β,and IL-10 were up-regulated in the brain tissues of zebrafish in the high-,medium-,and low-dosage groups of ZSSF compared those in the model group(P<0.001).Conclusions ZSSF exerts its antidepressant effect through multiple components and targets,and its antidepressant effects may be associated with its inhibition of inflammatory factors.
3.Hypoglycemic effect of Chinese yam polysaccharide on DEX-induced IR-3T3-L1 adipocytes
Xinyue LIU ; Fengzhong WANG ; Hanwen ZHENG ; ALBERTO Carlos Pires Dias ; Bei FAN ; Qiong WANG
Chinese Journal of Comparative Medicine 2024;34(4):73-83
Objective To examine the hypoglycemic effect of Chinese yam polysaccharide on the 3T3-L1 insulin resistance cell model.Methods IR-3T3-L1 adipocytes were randomly divided into control group(Con),model group(DEX),metformin group(Met,positive drug group)and CYPS group(0.05,0.15,0.45 mg/mL).After the creation of the IR-3T3-L1 cell model by dexamethasone(DEX)(1 μmol/L)stimulation,Chinese yam polysaccharide treatment was applied.Glucose intake and the levels of TC,TG,LDL-C,HDL-C,GSH-Px,MDA,HK,and PK were then measured.AMPK-PI3K/Akt signaling pathway-associated gene expression was identified using qRT-PCR.Results The glucose consumption of IR-3T3-L1 cells was considerably decreased after 48 hours of treatment with 1 μmol/L DEX compared to that of the Con group(P<0.01),and the reduction persisted for 60 hours.0.05,0.15,0.45 mg/mL CYPS treatment significantly increased glucose consumption(P<0.01),HK,PK,GSH-Px enzyme activities(P<0.01),HDL-C content(P<0.01),and decreased MDA enzyme activity(P<0.01),T-CHO,TG,LDL-C content(P<0.01)in IR-3T3-L1 cells.01).Additionally,CYPS administration dramatically decreased PI3K,Akt,GLUT-4,AMPK,IRS-2,PPARa,and adiponectin mRNA expression levels(P<0.05)and reduced IRS-1,GSK-3β,ACC,and FAS mRNA expression levels(P<0.01).Conclusions In IR-3T3-L1 cells,CYPS can reduce oxidative stress,control lipid metabolism disorders,and enhance DEX-induced glucose intake.AMPK-PI3K/Akt signaling pathway-associated genes may be connected to the process.