OBJECTIVE:To observe the effects of moxibustion at Zusanli(ST36)on rats with chronic fatigue syndrome(CFS)and to analyze the mechanisms of moxibustion through hippocampal Proteomics.METHODS:Male Sprague-Dawley(SD)rats...OBJECTIVE:To observe the effects of moxibustion at Zusanli(ST36)on rats with chronic fatigue syndrome(CFS)and to analyze the mechanisms of moxibustion through hippocampal Proteomics.METHODS:Male Sprague-Dawley(SD)rats were randomly divided into three groups:control group(CON),model group(MOD),and moxibustion group(MOX),with 12 rats in each group.The MOD and MOX groups underwent chronic multi-factor stress stimulation for 35 d to establish the CFS model.After modeling,the rats in the MOX group received mild moxibustion at Zusanli(ST36)(bilateral)for 10 minutes daily for 28 d.During the treatment period,rats in both the MOD and MOX groups continued modeling,while the CON group was kept under normal breeding conditions.The general condition of the rats was monitored,and behaviors were assessed using the Open Field Test(OFT),Exhaustion Treadmill Test,and Morris Water Maze(MWM).Hematoxylin and eosin(HE)staining and transmission electron microscopy(TEM)were employed to observe morphological changes in the hippocampus.Label-free Proteomics were utilized to identify differentially expressed proteins(DEPs)in the hippocampus,followed by bioinformatics analysis.The reliability of the Proteomics results was verified using Parallel Reaction Monitoring.RESULTS:A:Moxibustion at Zusanli(ST36)significantly reduced the general condition score of CFS rats,improved their behavioral performance in OFT,treadmill and MWM,and repaired the pathological and synaptic structural damage in the hippocampus.B:We identified DEPs by applying a fold change threshold of 1.2 and a significance level of P<0.05.In the comparison between the CON and the MOD,we identified a total of 72 DEPs(31 up-regulated and 41 down-regulated)associated with the development of CFS.In the comparison between the MOX and the MOD group,we identified a total of 103 DEPs(40 up-regulated and 63 down-regulated)related to the therapeutic effects of moxibustion.Gene Ontology(GO)enrichment analysis showed that CFS and moxibustion treatment were related to multiple biological processes,molecular functions,and cellular components.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis revealed that CFS pathogenesis was linked to base excision repair,steroid biosynthesis,and systemic lupus erythematosus,Furthermore,the treatment of CFS with moxibustion was relevant to terpenoid skeleton biosynthesis.C:Compared with the two comparison groups,we identified 16 potential biomarkers,noting that moxibustion reversed the upregulation of 14 DEPs and the down-regulation of 2 DEPs in CFS.These proteins are mainly associated with synaptic plasticity,ribosomal function,neurotransmitter secretion,glycine metabolism,and mitochondrial function.CONCLUSION:Moxibustion at Zusanli(ST36)is effective in treating CFS,the potential biomarkers identified by Proteomics confirm that the mechanisms of moxibustion involve multiple targets and pathways,which may be key to regulating the structural and functional damage in the hippocampus associated with CFS,highlighting their significant value for future research.展开更多
基金National Natural Science Foundation of China projects:Research on the Mechanism of Moxibustion Activating Calcium/Calmodulin-dependent Protein KinaseⅡPhosphorylation Mediating Long-term Potentiation to Regulate Synaptic Plasticity in the Treatment of Chronic Fatigue Syndrome Cognitive Impairment(No.82305394)Research on the Mechanism of Electroacupuncture Regulating Murine Double Minute 2 Ubiquitination Postsynaptic Density Protein 95 Levels to Reshape the Synaptic Structure of Hippocampal Neurons and Improve Chronic Fatigue Syndrome Cognitive Impairment(No.82074539)+7 种基金Outstanding Youth Project of Heilongjiang Provincial Natural Science Foundation:the Mechanism of Moxibustion in Improving Chronic Fatigue Syndrome Cognitive Impairment by Regulating Calcium/Calmodulin-dependent Protein Kinase II-mediated Long-Term Potentiation(No.YQ2023H019)Youth Talent Support Project of the Chinese Association of Traditional Chinese Medicine:Research on the Molecular Pathways of Moxibustion at the Zusanli(ST36)Point in Regulating Synaptic Plasticity to Improve Chronic Fatigue Syndrome Cognitive Impairment(No.2023-QNRC2-A04)China Postdoctoral Science Foundation:Research on the Mechanism of Moxibustion in Regulating Mitochondrial Autophagy Mediating Microglial Polarization to Promote Synaptic Remodeling in Chronic Fatigue Syndrome Cognitive Impairment(No.2024MD763980)Post-doctoral Program of Heilongjiang Province:based on the Calcium/Calmodulin-dependent Protein Kinase II/Parkin/NLR Family Pyrin Domain Containing 3 Signaling Axis to Explore the Mechanism of Moxibustion in Mitochondrial Autophagy Mediating Microglial Polarization to Promote Synaptic Remodeling in Chronic Fatigue Syndrome Cognitive Impairment(No.LBH-TZ2420)Study on the Mechanism of Moxibustion at the Zusanli(ST36)Regulating Long-Term Potentiation to Promote Synaptic Remodeling and Improve Cognitive Impairment in Chronic Fatigue Syndrome(No.LBH-Z23281)Chunhui Plan of the Ministry of Education:Study on the Mechanism of the Tongdu Yu Pi Tiaoshen Acupuncture in Improving Hippocampal Synaptic Plasticity of Chronic Fatigue Syndrome Cognitive Impairment(No.HZKY20220308-202201357)Youth Talent Support Project of the Heilongjiang Provincial Association of Traditional Chinese Medicine:Study on the Effect and Mechanism of Tongdu Yupi Tiaoshen Acupuncture on Improving Hippocampal Synaptic Plasticity in Chronic Fatigue Syndrome Cognitive Impairment(No.2022-QNRC1-05)Research Project of Traditional Chinese Medicine in Heilongjiang Province:Study on the Mechanism of Acupuncture in Improving Hippocampal Synaptic Plasticity of Chronic Fatigue Syndrome Cognitive Impairment(No.ZHY2022-136)。
文摘OBJECTIVE:To observe the effects of moxibustion at Zusanli(ST36)on rats with chronic fatigue syndrome(CFS)and to analyze the mechanisms of moxibustion through hippocampal Proteomics.METHODS:Male Sprague-Dawley(SD)rats were randomly divided into three groups:control group(CON),model group(MOD),and moxibustion group(MOX),with 12 rats in each group.The MOD and MOX groups underwent chronic multi-factor stress stimulation for 35 d to establish the CFS model.After modeling,the rats in the MOX group received mild moxibustion at Zusanli(ST36)(bilateral)for 10 minutes daily for 28 d.During the treatment period,rats in both the MOD and MOX groups continued modeling,while the CON group was kept under normal breeding conditions.The general condition of the rats was monitored,and behaviors were assessed using the Open Field Test(OFT),Exhaustion Treadmill Test,and Morris Water Maze(MWM).Hematoxylin and eosin(HE)staining and transmission electron microscopy(TEM)were employed to observe morphological changes in the hippocampus.Label-free Proteomics were utilized to identify differentially expressed proteins(DEPs)in the hippocampus,followed by bioinformatics analysis.The reliability of the Proteomics results was verified using Parallel Reaction Monitoring.RESULTS:A:Moxibustion at Zusanli(ST36)significantly reduced the general condition score of CFS rats,improved their behavioral performance in OFT,treadmill and MWM,and repaired the pathological and synaptic structural damage in the hippocampus.B:We identified DEPs by applying a fold change threshold of 1.2 and a significance level of P<0.05.In the comparison between the CON and the MOD,we identified a total of 72 DEPs(31 up-regulated and 41 down-regulated)associated with the development of CFS.In the comparison between the MOX and the MOD group,we identified a total of 103 DEPs(40 up-regulated and 63 down-regulated)related to the therapeutic effects of moxibustion.Gene Ontology(GO)enrichment analysis showed that CFS and moxibustion treatment were related to multiple biological processes,molecular functions,and cellular components.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis revealed that CFS pathogenesis was linked to base excision repair,steroid biosynthesis,and systemic lupus erythematosus,Furthermore,the treatment of CFS with moxibustion was relevant to terpenoid skeleton biosynthesis.C:Compared with the two comparison groups,we identified 16 potential biomarkers,noting that moxibustion reversed the upregulation of 14 DEPs and the down-regulation of 2 DEPs in CFS.These proteins are mainly associated with synaptic plasticity,ribosomal function,neurotransmitter secretion,glycine metabolism,and mitochondrial function.CONCLUSION:Moxibustion at Zusanli(ST36)is effective in treating CFS,the potential biomarkers identified by Proteomics confirm that the mechanisms of moxibustion involve multiple targets and pathways,which may be key to regulating the structural and functional damage in the hippocampus associated with CFS,highlighting their significant value for future research.
