Active fraction combination from Liuwei Dihuang decoction improves adult hippocampal neurogenesis and neurogenic microenvironment in cranially irradiated mice

OBJECTIVE Cranial radiotherapy is clinically used in the treatment of brain tumors;however,the consequent cognitive and emotional dysfunctions seriously impair the life quality of patients.LW-AFC,an active fraction combination extracted from classical traditional Chinese medicine prescription Liuwei Dihuang decoction,can improve cognitive and emotional dysfunctions in many animal models;however,the protective effect of LW-AFC on cranial irradiation-induced cognitive and emotional dysfunctions has not been reported.Recent studies indicate that impairment of adult hippocampal neurogenesis(AHN)and alterations of the neurogenic microenvironment in the hippocampus constitute critical factors in cognitive and emotional dysfunctions following cranial irradiation.Here,our research further investigated the potential protective effects and mechanisms of LW-AFC on cranial irradiation-induced cognitive and emotional dysfunctions in mice.METHODS LW-AFC(1.6 g·kg^(-1))was intragastrically administered to mice for 14 d before cranial irradiation(7 Gyγ-ray).AHN was examined by quantifying the number of proliferative neural stem cells and immature neurons in the dorsal and ventral hippocampus.The contextual fear conditioning test,open field test,and tail suspension test were used to assess cognitive and emotional functions in mice.To detect the change of the neurogenic microenvironment,colorimetry and multiplex bead analysis were performed to measure the level of oxidative stress,neurotrophic and growth factors,and inflammation in the hippocampus.RESULTS LW-AFC exerted beneficial effects on the contextual fear memory,anxiety behavior,and depression behavior in irradiated mice.Moreover,LW-AFC increased the number of proliferative neural stem cells and immature neurons in the dorsal hippocampus,displaying a regional specificity of neurogenic response.For the neurogenic microenvironment,LW-AFC significantly increased the contents of superoxide dismutase,glutathione peroxidase,glutathione,and catalase and decreased the content of malondialdehyde in the hippocampus of irradiated mice,accompanied by the increase in brain-derived neurotrophic factor,insulin-like growth factor-1,and interleukin-4 content.Together,LW-AFC improved cognitive and emotional dysfunctions,promoted AHN preferentially in the dorsal hippocampus,and ameliorated disturbance in the neurogenic microenvironment in irradiated mice.CONCLUSION LW-AFC ameliorates cranial irradiation-induced cognitive and emotional dysfunctions,and the underlying mechanisms are mediated by promoting AHN in the dorsal hippocampus and improving the neurogenic microenvironment.LW-AFC might be a promising therapeutic agent to treat cognitive and emotional dysfunctions in patients receiving cranial radiotherapy.

Effects of LW-AFC,a new formula derived from Liuwei Dihuang decoction,on intestinal microbiome in senescence-accelerated mouse prone 8 strain,a mouse model of Alzheimer disease

OBJECTIVE To investigate the effects of LW-AFC,a new formula derived fromLiuwei Dihuang decoction,on gut microbiota and the behavior of learning and memory of SAMP8 mice,a mouse model of Alzheimer Disease(AD),and identify the specific intestinal microbiota correlating with cognitive ability.METHODS Morris-water maze test,novel object recognition test and shuttle-box test were conducted to observe the ability of learning and memory.16S rRNA amplicon sequencing(Illumina,San Diego,CA,USA)was employed to investigate gut microbiota.RESULTS The treatment of LW-AFC improved cognitive impairments of SAMP8 mice,including spatial learning and memory ability,active avoidance response,and object recognition memory capability.Our data indicated that there were significantly 8 increased and 12 decreased operational taxonomic units(OTUs)in the gut microbiota of SAMP8 mice compared with senescence accelerated mouse resistant 1(SAMR1) strains,the control of SAMP8 mice.The treatment of LW-AFC altered 22(16 increased and 6 decreased)OTUs in SAMP8 mice and among them,15 OTUs could be reversed by LW-AFC treatment resulting in a microbial composition similar to that of SAMR1 mice.We further showed that there were7(3 negative and 4 positive correlation)OTUs significantly correlated with all the three types of cognitive abilities,at the order level,including Bacteroidales,Clostridiales,Desulfovibrionales,CW040,and two unclassified orders.LW-AFC had influences on bacterial taxa correlated with the abilities of learning and memory in SAMP8 mice and restored them to SAMR1 mice.CONCLUSION The effects of LW-AFC on improving cognitive impairments of SAMP8 mice might be via modulating intestinal microbiome and LW-AFC could be used as a potential anti-AD agent.

Pharmacological mechanism of Liuwei Dihuang Decoction in treating Osteoporosis and Alzheimer’s disease based on Network Pharmacology

Objective:To predict the active components,targets of Liuwei Dihuang Decoction for the treatment of osteoporosis and Alzheimer’s disease with network pharmacology analysis.Methods:The components and targets of all the herbs of Liuwei Dihuang Decoction were obtained from ETCM database,Osteoporosis and Alzheimer’s disease targets were extracted from the GeneCards and DrugBank databases,selected based on STRING database for PPI network construction and build the network through the software of Cytoscape,Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses were carried out by R software.Results:181 active compounds and 762 disease-related from Liuwei Dihuang Decoction,there were 144 disease-related common targets obtained from Venn Diagram.IL6,TNF,PPARA,PPARG,SRC and other 11 targets are the node protein of the whole network.In the multisystem biological reaction process such as neurotransmitter,glucose and lipid metabolism,calcium and phosphorus metabolism.Conclusion:Liuwei Dihuang pill may play a co-therapeutic role in Osteoporosis and Alzheimer’s disease by participating in the multi-system biological reaction process such as neurotransmitter,glucose and lipid metabolism,calcium and phosphorus metabolism.

Shengmaisan combined with Liuwei Dihuang Decoction alleviates chronic intermittent hypoxia-induced cognitive impairment by activating the EPO/EPOR/JAK2 signaling pathway

Chronic intermittent hypoxia(CIH),a principal pathophysiological aspect of obstructive sleep apnea(OSA),is associated with cognitive deficits.Clinical evidence suggests that a combination of Shengmaisan and Liuwei Dihuang Decoctions(SMS-LD)can enhance cognitive function by nourishing yin and strengthening the kidneys.This study aimed to assess the efficacy and underlying mechanisms of SMS-LD in addressing cognitive impairments induced by CIH.We exposed C57BL/6N mice to CIH for five weeks(20%-5%O_(2),5 min/cycle,8 h/day)and administered SMS-LD intragastrically(15.0 or 30 g·kg^(-1)·day)30 min before each CIH session.Additionally,AG490,a JJanus kinase 2(JAK2)inhibitor,was administered via intracerebroventricular injection.Cognitive function was evaluated using the Morris water maze,while synaptic and mitochondrial structures were examined by transmission electron microscopy.Oxidative stress levels were determined using DHE staining,and the activation of the erythropoietin(ER)/ER receptor(EPOR)/JAK2 signaling pathway was analyzed through immunohistochemistry and Western blotting.To further investigate molecular mechanisms,HT22 cells were treated in vitro with either SMS-LD medicated serum alone or in combination with AG490 and then exposed to CIH for 48 h.Our results indicate that SMS-LD significantly mitigated CIH-induced cognitive impairments in mice.Specifically,SMS-LD treatment enhanced dendritic spine density,ameliorated mitochondrial dysfunction,reduced oxidative stress,and activated the EPO/EPOR/JAK2 signaling pathway.Conversely,AG490 negated SMS-LD’s neuroprotective and cognitive improvement effects under CIH conditions.These findings suggest that SMS-LD’s beneficial impact on cognitive impairment and synaptic and mitochondrial integrity under CIH conditions may predominantly be attributed to the activation of the EPO/EPOR/JAK2 signaling pathway.

Effects of Liuwei Dihuang Decoction(Yukmijihwang-tang)on Physical Fatigue by Regulating Neurotransmitters in Brain

Objective:To study the effect of Liuwei Dihuang Decoction(六味地黄汤)or Yukmijihwangtang(YJT)on endurance exercise by in vivo experiment.Methods:ICR mice were randomly divided into the control group(distilled water)and the YJT groups(1,10,100 mg/kg),5 animals per group.YJT and distilled water were orally administered.The anti-fatigue effect of YJT was evaluated by open fifiled test(OFT),forced swimming test(FST),and tail suspension test(TST).Results:In the OFT,YJT signifificantly increased the total movement distance in a dose-dependent manner.Additionally,treatment with YJT signifificantly decreased immobility time in the FST and the TST.Various neurotransmitters such as norepinephrine(NE),serotonin(5-HT),dopamine(DA)levels were increased by FST.Administration of YJT down-regulated the expression levels of NE,5-HT,5-hydroxyindole-acetic acid(5-HIAA),and DA in the brain stem and hypothalamus of mice.Moreover,protein expression of HSP70 in mice liver and heart muscles was signifificantly increased in the YJT groups.Conclusions:YJT could ameliorate fatigue and enhance exercise tolerance through suppressing of brain monoamines including NE,5-HT,5-HIAA,and DA in FST mice model.

Effect of Liuwei Dihuang Decoction (六味地黄汤) on LongTerm Potentiation in Hippocampal Dentate Gyrusin Rats

Objective: To investigate effect of Liuwei Dihuang decoction (LWDHD), a traditional Chinese medicinal prescription, on the formation of long term potentiation (LTP) in the hippocampal dentate gyrus in anesthetized rats. Methods: LTP experiments were conducted by using a bipolar stimulating electrode to stimulate perforant pathway and a monopolar electrode to record the evoked field potential in hippocampal dentate gyrus. In the present study, both suprathreshold and subthreshold tetanic stimulation to the perforant pathway were employed. Results: Single oral administration of LWDHD (per os, 5 g/kg and 10 g/kg respectively) has no effect on the basal spike amplitude. However, LWDHD (per os, 5 g/kg and 10 g/kg respectively) significantly intensified the spike amplitude induced by a subthreshold stimulation (60 Hz, 20 pulses) while it did not affect formation of LTP formation induced by suprathreshold tetanic stimulation (60 Hz, 30 pulses). Conclusion: The present results suggested that LWDHD given orally potentiate the synaptic plasticity and transmission in hippocampal dentate gyrus, which might account for the enhancing effect of LWDHD on learning and memory in senescence accelerated mice.

LW-AFC,a new traditional Chinese medicine formula,shows promising therapeutic effects on Alzheimer disease

Alzheimer disease(AD) is the leading cause of dementia that affects millions of elderly people worldwide.The currently available therapies have limited efficacy. Liuwei Dihuang decoction(LW),a classical traditional Chinese medicine(TCM) formula,has long been used to treat various diseases,including dementia. A large number of pharmacological studies have showed that LW has beneficial effects on AD. LW-AFC is a new formula consisted of the main active components prepared fromLW.In this study,we found that administration of LW-AFC significantly improved behavioral performances in spontaneous locomotor activity,object recognition memory,spatial learning and memory,passive and active avoidance impairment in some AD mouse models,such as SAMP8 and APP/PS1 transgenic mice. Meanwhile,the impairments of long-term potentiation(LTP) were significantly ameliorated by LW-AFC administration in SAMP8 mice,as well as in corticosterone-induced LTP inhibition mice model.These effects were companied by alleviating of the neuron loss in the hippocampus,suppressing of A-beta deposition in the brain,and reducing of the concentration of Abeta42 in the hippocampus and plasma of APP/PS1 mice. The restoring and correcting of imbalance of hypothalamic-pituitary-adrenal(HPA) and hypothalamic-pituitary-gonadal(HPG) axis,the disorder of lymphocyte subsets,the abnormality of production of cytokine were also observed in LW-AFC treated SAMP8 and APP/PS1 mice.These findings indicated that LW-AFC ameliorated the behavioral and pathological deterioration of AD mice via the restoration of the NIM network in a holistic way,suggesting that LW-AFC might bea promising therapy for AD.