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.

Research on Xijiao Dihuang Decoction suppressing platelet apoptosis in immune-mediated aplastic anemia based on mitochondrial mediated pathway

The aplastic anemia mice model was established in this study.BALB/c mice shall be treated with whole body irradiation with 60 Co-γradiation(5.5Gy,1.1Gy/min×5 min).then within 4 h,DBA/2 mice were injected lymphocyte suspension 1×106 cells/mouse through caudal vein.Grouping of testing animals:Normal control mice were healthy C57BL/6 mice without AA modeling.AA control group mice were exposed to radiation and cell transfusion and had no treatment with either CSA or XDD.CSA group mice received daily lavage with 0.027g/kg(0.1ml/10g)of CSA whereas XDD group mice received daily lavage with 19.5g/kg of XDD.The experimental result indicated CSA and XDD lavage mice had significantly higher platelet count andΔΨm than AA mice(P<0.05).Levels of Cyt C,PS and Ca2+were significantly lower in CSA and XDD groups compare with AA group.More specifically,compared to CSA group,XDD group also had lower level ofΔΨm and higher level of Cyt C and Ca2+(both P<0.05).Both CSA and XDD treatment reduced Bak and Bax levels significantly compared to AA mice.However,XDD treatment still showed higher expressions than CSA(P<0.05).CSA and XDD treatment increased the levels of Pro-apoptotic protein expressions(caspase-8,caspase-3,caspase-9).XDD showed less potent effect than CSA in increasing these protein expressions(all P<0.05).Therefore,we hypothesized XDD was possible to increase platelet number and prevent its apoptosis in immune-induced AA via the mitochondrial pathway.

Meta-analysis of the correlation between diabetic nephropathy and inflammatory response based on Shenqi Dihuang Decoction

Objective:A meta-analysis was used to systematically study the correlation between inflammatory response and diabetic nephropathy based on Shenqi Dihuang Decoction.Methods:The RCT literatures related to the efficacy and inflammatory response of Shenqi Dihuang Decoction in the treatment of diabetic nephropathy in Chinese and English databases were searched.The time limit was from the establishment of the database to February 2020.Literature screening and data extraction were performed according to the criteria for inclusion and exclusion.The evaluator's manual 5.1.0 was used to evaluate the risk of bias in the final included literature.Meta-analysis was performed using RevMan 5.3 software.Comparing western medicine treatment and Shenqi Dihuang decoction to inflammatory factors:interleukin-6(IL-6)Interleukin-8(IL-8),tumor necrosis factor-α(TNF-α),C-reactiveprotein(CRP),intercellular adhesion molecule-1(MCP-1),and monocyte chemotactic protein-1(ICAM-1)and efficacy indicators:the impact of urine microalbumin(mALB)and clinical efficacy.Results:A total of 11 related research reports were finally included,of which 9 evaluated IL-6[SMD=-1.09,95%CI(-1.57,-0.61),Z=4.43,P<0.00001];5 Item evaluated IL-8[SMD=-0.91,95%CI(-1.44,-0.37),Z=3.33,P=0.0009<0.05];7 items evaluated TNF-[SMD=-0.97,95%CI(-1.49,-0.45),Z=3.63,P=0.0003<0.05];9 items evaluated CRP[SMD=-1.40,95%CI(-1.89,-0.90),Z=5.49,P<0.00001];2 items evaluated MCP-1[SMD=-0.71,95%CI(-1.00,-0.43),Z=4.95,P<0.00001];1 item was ICAM-1 Evaluation was performed[SMD=-0.98,95%CI(-1.38,-0.58),Z=4.82,P<0.00001];6 items used mALB as the efficacy index[SMD=-1.85,95%CI(-2.65,-1.05),Z=4.52,P<0.00001];9 items evaluated the clinical efficacy,[OR=4.45,95%CI(2.96,6.69),Z=7.17,P<0.00001].Conclusion:Shenqi Dihuang Decoction was used in patients with diabetic nephropathy.Compared with the western medicine treatment group,the levels of IL-6,IL-8,TNF-α,CRP,MCP-1,ICAM-1and inflammatory factors were significantly reduced.The effect is significantly increased.It is suggested that Shenqi Dihuang Decoction has better anti-inflammatory response than diabetic nephropathy than western medicine alone.

Xijiao Dihuang Decoction combined with Yinqiao Powder reverses influenza virus-induced F-actin reorganization in PMVECs by inhibiting ERM phosphorylation

Objective:It has been documented that ezrin/radixin/moesin(ERM)phosphorylation by the p38 mitogen-activated protein kinase(MAPK),Rho/ROCK,and protein kinase C(PKC)pathways leads to filamentous actin(F-actin)reorganization and microvascular endothelial cell hyperpermeability.In this study,we investigated the effects of Xijiao Dihuang Decoction combined with Yinqiao Powder(XDY)on influenza virus(IV)-induced F-actin restructuring and ERM phosphorylation regulated by the Rho/Rho kinase 1(ROCK),p38 MAPK,and PKC signaling pathways in pulmonary microvascular endothelial cells(PMVECs).Methods:Serum containing XDY(XDY-CS;13.8 g/kg)was acquired using standard protocols for serum pharmacology.Primary PMVECs were obtained from male Wistar rats and cultured.After adsorption of IV A(multiplicity of infection,0.01)for 1 h,medium with 20%XDY-CS was added to the PMVECs.The distributions of F-actin and phosphorylated ERM were determined by confocal microscopy,and F-actin expression was measured by flow cytometry.The expression levels of ROCK1,phosphorylated myosin phosphatase target-subunit(p-MYPT),phosphorylated MAPK kinase,phosphorylated p38(p-p38),phosphorylated PKC(p-PKC),and phosphorylated ERM(p-ERM)were determined by western blotting.Results:F-actin reorganization in IV-infected PMVECs was reversed by XDY-CS treatment,which was accompanied by reduced p-ERM production.The p-ERM protein accumulated at plasma membrane of PMVECs infected with IV,which was also inhibited by XDY-CS treatment.

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.

Effect of Modified Xijiao Dihuang Decoction (加味犀角地黄汤) on Intestinal Flora and Th17/Treg in Rats with Radiation Enteritis

Objective To observe the effect of Modified Xijiao Dihuang Decoction(加味犀角地黄汤,MXDD)on rats with radiation enteritis,and explore its action mechanism.Methods Thirty female Sprague Dawley rats were divided into the control,model,dexamethasone(DXM),golden bifid(GB)and MXDD groups using random number table,6 rats in each group.Except the control group,the other rats were developed into radiation enteritis model by exposing to a single 60Co-γray at a dose of 11 Gy.The rats in the DXM,GB and MXDD groups were treated with DXM(1.425 mg/kg),GB(0.8 g/kg)and MXDD(36.0 g/kg)for 3 days,respectively.Body weight and diarrhea condition of rats were evaluated daily.On day 3,the feces of rats were collected for intestinal flora detection and the small intestinal tissues were also collected.Bacterial species annotation,alpha and beta diversities as well as composition of intestinal flora were detected and compared.The protein and mRNA expressions of interleukin 17(IL-17),retinoid-related orphan nuclear receptor gamma t(ROR-γt)and forkhead/winged helix transcription factor p3(FoxP3)were determined by Western blot and polymerase chain reaction,respectively.The abundance and diversity of intestinal flora as well as the proportion at the phylum and genus levels were assayed by 16S rRNA metagenome sequencing.Correlation between intestinal flora and Th17/Treg was analyzed by heatmap method.Results On day 1 to 3 after radiation,compared with the control group,the body weight in model group was decreased(P<0.05 or P<0.01).Compared with the model group,MXDD could alleviate weight loss and diarrhea caused by irradiation.At the phylum level,MXDD cause a significant increase in Firmicutes,and a decrease in Proteobacteria(P<0.05 or P<0.01).At the genus level,MXDD reduced the proportion of Escherichia Shigella(P<0.01).In addition,IL-17 and FoxP3 mRNA and protein expression levels were down-regulated and ROR-γt was up-regulated by MXDD treatment(P<0.05).Besides,Firmicutes and Lactobacillus were positively correlated with FoxP3(r=0.73,0.79,respectively;P<0.01),negatively correlated with IL-17(r=0.66,0.64,respectively;P<0.01 or P<0.05)and ROR-γt(r0.73,0.81,respectively;P<0.01).Proteobacteria and Escherichia Shigella both had positive correlation with IL-17(r 0.77,0.57,respectively;P<0.01 or P<0.05)and ROR-γt(r=0.94,0.79,respectively;P<0.01)and negative correlation with FoxP3(r0.74,0.65;P<0.01).Conclusion MXDD could improve the survival status of irradiated rats by regulating the richness,diversity and composition of intestinal flora,and restoring the balance of Th17/Treg.

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.

Effect of modified Dihuang Yinzi recipe and Huatan Tongluo decoction on neurological function of MCAO rats

Objective To observe the effect of Modified Dihuang Yinzi Recipe(MDYR)and Hua-tan Tongluo Decoction(HTD)on neurological function of middle cerebral artery occlusion(MCAO)model rats.Methods Forty SD rats were randomly divided into 5 groups,i.e.,the sham-op-

Clinical Observation of the Treatment of Exudative Central Chorioretinitis with Traditional Chinese Medicine

Objective: To observe the curative effects of Jiajian Huaban decoction ( ) and Zhibai Dihuang decoction ( ) in the treatment of exudative central chorioretinitis.Methods: Forty-nine eyes (47 patients) with exudative central chorioretinitis were treated with Jiajian Huaban Decoction in the period of active stage of inflammation and with Zhibai Dihuang Decoction in the period of retrograde stage. The curative effects were evaluated according to fluorescin fundus angiography and their visual functions.Results: The average period of treatment was 4 months. Forty-three eyes among the 49 were effective and the effective rate was 87.75%.Conclusion: Jiajian Huaban Decoction and Zhibai Dihuang Decoction could promote visual acuity and shorten the period of disease in the treatment of exudative central chorioretinitis.