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 Liuwei Dihuang decoction and its compatible prescriptions on the function of hypothalamus-pituitary-adrenal axis in senescence-accelerated mouse

OBJECTIVE To investigate age-related functional change of hypothalamus-pituitary-adrenal(HPA)axis in senescence accelerated mouse(SAM)and the effects of Liuwei Dihuang decoction(LW)and its San-bu(three tonics)and San-xie(three eliminators)components on the function of HPA axis.METHODS Male SAM-resistance/1(SAMR1)and SAM-prone/8(SAMP8)at the ages of 6and 12 months old were used.SAMP8 were orally administered with LW,three tonics and three eliminators at the doses of 10,6.4and 3.6g·kg-1·d-1,respectively,for consecutive 60 d.Serum level of CORT was assayed with ELISA method.The levels of hypothalamic CRH and pituitary ACTH was determined with radioimmunoassay.RESULTS The levels of hypothalamic CRH,pituitary ACTH and serous CORT were much higher in 6 and 12 months old SAMP8 than those in age-matched SAMR1,which suggested the abnormal function of HPA axis in SAMP8.Oral administration of LW and three tonics significantly decreased the level of hypothalamic CRH of 6 and 12 months old SAMP8,and reduced the levels of pituitary ACTH and serous CORT of 6 month old SAMP8.Three eliminators significantly decreased the level of hypothalamic CRH of 6 months old SAMP8.The results indicated that oral administration of LW,three tonics and three eliminators improved the function of HPA axis of SAMP8.CONCLUSION The results showed the hyperactivity of HPA axis of SAMP8,and LW improved the hyperactivity of 6 month old SAMP8.Three tonics and three eliminators had similar effects as LW,which had better effect after compatibility.

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.

Clinical Observation and Network Pharmacology Study on Analgesic Effect of Qianghuo Chushi Decoction on Fasciitis

[Objectives]To observe the clinical analgesic effect of Qianghuo Chushi Decoction(QHCSD)on patients with fasciitis,and explore its possible molecular mechanism based on network pharmacology.[Methods]120 enrolled patients were randomly divided into experimental group and control group,and were separately treated with QHCSD formula granules and Diclofenac Sodium Enteric-coated Tablets for 4 weeks.The patient’s pain visual analogue scale(VAS)was used as the curative effect indicator.The molecular action mechanism of QHCSD was predicted based on network pharmacology,the active components of QHCSD were screened using TCMSP database,potential targets were predicted by PharmMapper server,compound-target network and protein interaction network were constructed,and GO-based enrichment analysis and KEGG-based biological pathway enrichment analysis were performed.[Results]After treatment,the pain scores in each group were significantly lower than those before treatment(P<0.01),the score of the experimental group was significantly lower than that of the control group(P<0.01),and the total effective rate of the experimental group was 83.33%,which was significantly higher than that of the control group(78.33%,P<0.05).Based on 108 active components in QHCSD,a compound-target network was constructed.The PPI network contained 155 nodes and 527 interaction relationships,and key nodes included FOS,ESR1,NCOA1,RELA,EGFR,MAPK8,IL-6,etc.The GO pathway mainly involved steroid hormone and its receptor activity,RNA polymerase II transcriptional regulator binding,nuclear receptor activity,protein heterodimerization activity and other pathways.KEGG metabolic pathways included PI3 K-Akt signaling pathway,Kaposi’s sarcoma-associated herpesvirus(KSHV)infection and other pathways.[Conclusions]QHCSD has a significant analgesic effect on fasciitis,and the PI3 K-Akt signaling pathway may be the key pathway for its analgesic effect.

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.

Research of the anxiolytic effect of Baihe Dihuang Decoction

Objective:To study the effect of Baihe Dihuang Decoction in improving anxiety and its mechanism of action.Methods: Male mice were randomly divided into 5 groups: blank control group, Baihe Dihuang Decoction high, medium, low-dose group and diazepam group. After continuous intragastric administration for 14 d, the behavioral test of the mouse elevated plus maze experiment and opening test was performed. The brain tissue GABA and Glu content was measured by immunosorbent assay (ELISA).Results: Compared with the blank control group, the middle and high doses of Baihe Dihuang Decoction could increase the time and frequency of movement of the mice in the open cross labyrinth in the open arm and increase the number of times that the mouse entered the central area in the opening test. The anti-anxiety effect also showed an increase in the brain tissue GABA content in mice, ignificantly decreased Glu contents in mice.Conclusion: Baihe Dihuang Decoction has some anxiolytic effect, and anti-anxiety effect may be related to increasing brain tissue GABA content, decreased Glu contents in mice.

Construction of NIM sub-network regulated by Liuwei Dihuang decoction and its characteristics

OBJECTIVE To construct the neuroendocrine immunomodulation(NIM) sub-network regulated by Liuwei Dihuang decoction(LW) and analyze its characteristics.METHODS We took the GSE57273 in GEO database and screened the differentially expressed genes(DEGs)(P<0.01) by the GEO2 R tool as gene expression signature of LW.The global PPI network was constructed in the context of whole PPI network through direct interaction algorithm and forest algorithm respectively.Then the enrichment and the topological characteristics of NIM signaling molecules were evaluated by permutation test.Finally,we abstracted the NIM sub-network by NIMNT,which combined the NIM molecular network and forest algorithm,and analyzed the topological characteristics of it by the Network Analyzer(release 2.7) plugin in Cytoscape v3.5.1.RESULTS We got 2468 DEGs in the gene expression signature of LW.After analyzing the global PPI network of LW got by two kinds of algorithms,we found that the NIM signaling molecules significantly enriched and located in important positions in the global PPI network.The NIM sub-network regulated by LW contained 1099 nodes and 1056 edges.We screened out 22 hub nodes(Degree>10).Among them,there were 19 NIM signaling molecules in which only ESR1 changed significantly and 3 non-DEGs(NFATC2,RARA,TP53).However,the down.stream of the hub nodes were significantly changes.CONCLUSION The results suggested that LW might mainly regulate the non-hub nodes to recovery of the network imbalance of the body in the state of disease.

Liuwei Dihuang Decoction improves cognitive impairments via regulating immune system in APP/PS1 transgenic mouse, a mouse model of Alzheimer disease

OBJECTIVE To investigate the effect and mechanisms of Liuwei Dihuang Decoction(LW) on cognition in PrP-hAβPPswe/PS1ΔE9(APP/PS1) transgenic mice.METHODS LW was adminis.trated with oral for 3 months.The locomotor activity test was performed to investigate the spontaneous motor activity of mice.The Morris water maze test and shuttle box test were performed to investigate the spatial learning and memory and active avoidance response respectively.The Αβ deposits and neuron loss in the hippocampus was detected by immunofluorescence staining and nissl staining respectively.The flow cytometry was employed to investigate the lymphocyte subsets of the mice.The 3 H-thymidine incorporation was performed to investigate the splenocytes proliferation.RESULTS The treatment of LW ameliorated the impairments of spatial learning and memory and active and passive avoidance in APP/PS1 mice.The administration of LW alleviated neuron loss in the brain,suppressed amyloid-β(Αβ) deposits in the hippocampus of APP/PS1 mice.The treatment of LW significantly increased ConAand LPS-induced proliferation of splenocytes,increased CD3+ T cells and CD19+ B cells in the spleen lymphocytes and reduced Gr1+ cells in APP/PS1 mice.CONCLUSION This data indicated the adminis.tration of LW ameliorated behavioral and pathological deterioration via regulating immune function.

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.

梅花针配合首乌羌活汤治疗肝肾不足型斑秃临床观察

目的观察梅花针配合首乌羌活汤治疗肝肾不足型斑秃(AA)的临床效果。方法选取2020年1月—2020年6月大连市皮肤病医院门诊就诊的62例肝肾不足型AA患者作为研究对象,采用数字随机表法随机分为治疗组与对照组,各31例。对照组予口服首乌羌活汤治疗,治疗组在此基础上联用梅花针疗法,均以5周为1疗程,连续2个疗程。比较两组患者1个疗程和2个疗程后的效果。结果1个疗程后,治疗组总有效率为58.06%(18/31),高于对照组的25.81%(8/31)(P<0.05);2个疗程后,治疗组总有效率为83.87%(26/31),亦高于对照组的48.39%(15/31)(P<0.05)。结论梅花针配合首乌羌活汤治疗肝肾不足型AA,总有效率高、不良反应少,具有较好的临床效果。

Comparative pharmacokinetics of six components in normal and rheumatoid arthritis rats after intragastrical administration of Qianghuo Shengshi Decoction granules by LC-MS/MS

Objective:To investigate the plasma pharmacokinetics of six representative components(nodakenin,osthole,5-O-methylvisammioside,ferulic acid,liquiritigenin,and liquiritin),which were the ingredients of Qianghuo Shengshi Decoction(QSD)granules,in normal and rheumatoid arthritis(RA)rats administrated QSD granules intragastrically.Methods:A rapid and accurate ultra-high performance liquid chromatography-tandem mass spectrometry(LC-MS/MS)method was developed for the simultaneous determination of six components in plasma,and it showed a good specificity,linearity,intra-day and inter-day precision,intra-day and inter-day accuracy,extraction recovery,stability,and the less matrix effect.Results:The validated LC-MS/MS method was successfully used to compare the plasma pharmacokinetics of six ingredients between normal and RA rats after intragastrical administration of QSD granules and differences in the pharmacokinetics were found in two types of rats.The absorption rate in the RA rats was lower for nodakenin,osthole,5-O-methylvisammioside,liquiritigenin and liquiritin than in the normal group,while the absorption rate of ferulic acid remained constant in two groups.In comparison with the normal rats,the exposure concentration of nodakenin was higher and that of other five components except for nodakenin was lower under pathological conditions.Additionally,the absorptive amount of nodakenin,osthole,5-O-methylvisammioside and liquiritin was increased and that of ferulic acid and liquiritigenin was reduced in the RA rats than in the normal rats.Compared with the normal rats,the retention time of nodakenin,ferulic acid and liquiritin was reduced in vivo,whereas the retention time of osthole,5-O-methylvisammioside and liquiritigenin was raised in the body for the RA rats.In contrast to the normal rats,the data demonstrated an increase in the elimination velocity of nodakenin and a decrease in the elimination velocity of the other five components except for nodakenin in the pathological state.Conclusion:This study showed that the pharmacokinetic behavior of the six components,nodakenin,osthole,5-O-methylvisammioside,ferulic acid,liquiritigenin,and liquiritin,is different in vivo between normal and pathological states of rats,and this research provided the necessary experimental data to explain the pharmacokinetics of QSD granules in both normal and pathological states and provide some references for its clinical application at some level.

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.

The components transitive regularity of three dosage forms of Liuwei Dihuang Fufang

Objective To explore the transitive regularity of holistic constituents from the crude slices of the medicinal raw materials(MCS)to the formula granules(FG),fufang decoction(FD),and finally,the concentrated pills(CP)of Liuwei Dihuang Fufang(六味地黄复方,LWDHF).Methods Samples for MCS,FG,FD,and CP of LWDHF were obtained,and a fingerprint data-base was established using high-performance liquid chromatography(HPLC),by separating the samples in an XB-C18 column and analyzing the transitive regularity of components us-ing the total quantum statistical moment(TQSM),including total quantum zero moment(AUCT),total quantum first moment(MRTT),total quantum second moment(VRTT),and its similarity approach.The AUCT,MRTT,and VRTT were calculated based on the representative HPLC chromatograms of FG,FD,and CP of LWDHF.Results AUCT of FG,FD,and CP of LWDHF was 71804,46553,and 144646μV·s,respectively;MRTT was 14.43,14.54,and 18.85 min,respectively;and VRTT was 106.98,112.84,and 269.12 min2,respectively.Comparing the similarity of FG/FD,FG/CP and FD/CP of LWDHF,the TQSM similarity values were 98.66%,76.62%,and 75.37%,respectively,whereas the tradi-tional similarity evaluation values were 98.68%,85.43%,and 85.60%,respectively.Conclusion The results perform little distinction in the total composition between FG and FD,whereas some distinction existed between FD and CP.Experimental evidence,therefore indicates that FG could be used as the alternative of MCS in clinical applications.

基于生物信息学探讨犀角地黄汤治疗银屑病的作用机制

目的运用生物信息学方法筛选银屑病炎症相关的特征基因并探索犀角地黄汤的干预机制。方法从GEO数据库中筛选包含银屑病皮损及正常对照的数据集。从GSEA数据库中获得炎症相关基因。使用limma包筛选出差异表达基因并与炎症相关基因取交集获取差异表达的炎症相关基因(differentially expressed inflammation-related genes,DEIRGs),并对DEIRGs进行基因本体论(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。通过加权基因共表达网络分析(weighted geneco-expression network analysis,WGCNA)和最小绝对值收敛和选择算子(least absolute shrinkage and selection operator,LASSO)回归筛选出银屑病炎症相关的特征基因,并进行受试者工作特征曲线(receiver operating characteristic curve,ROC)检验。检索TCMSP、TCMID数据库,筛选犀角地黄汤活性成分和靶点,与DEIRGs取交集得到有效作用成分和有效靶点。然后,通过Pubchem、PDB数据库获取相关结构文件,再利用swissdock数据库进行分子对接并计算结合能。结果本研究共筛选560个差异表达基因和20个DEIRGs。GO和KEGG富集分析共得到779条生物功能和37条通路。WGCNA分析共筛选出483个基因,与20个DEIRGs取交集得到9个银屑病核心炎症反应基因,通过LASSO回归筛选出6个与银屑病发病高度相关的炎症反应特征基因,分别为CXCL10、F3、NMI、P2RY2、RTP4和TPBG。ROC曲线显示各个基因的曲线下面积(area under curve,AUC)均>0.9,LASSO回归模型的AUC为1。将犀角地黄汤的中药成分采用网络药理学检索解析后发现赤芍中的活性成分鞣花酸(ellagic acid)可作用于CXCL8,芍药苷(paeoniflorin)可作用于IL-6;牡丹皮中的活性成分栎精(quercetin)可作用于CXCL8、IL-6、F3和CXCL10。结论CXCL10、F3、NMI、P2RY2、RTP4和TPBG可能是银屑病的炎症相关特征基因,预测犀角地黄汤中的赤芍、牡丹皮有作用于银屑病相关特征基因的成分,可为后续药物研究提供参考。

加味百合地黄汤对围绝经期抑郁症模型大鼠HPO轴和雌激素受体表达的影响

目的基于下丘脑-垂体-卵巢(hypothalamic-pituitary-ovarian,HPO)轴及雌激素受体(estrogen receptor,ER)探明加味百合地黄汤治疗围绝经期抑郁症的功效及潜在机制。方法将大鼠随机分为正常对照组、模型组、阳性药物组(氟哌噻吨美利曲水溶液1.89 mg/kg)以及加味百合汤高、中、低剂量组(16.2 g/kg、8.1 g/kg、4.05 g/kg),每组10只。除正常对照组外,其他组均采用双侧卵巢摘除联合慢性不可预见性应激建立围绝经期抑郁症大鼠模型。各组给药21 d后采用糖水偏好实验和强迫游泳实验评价大鼠抑郁样行为,酶联免疫吸附法检测血清促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)、卵泡刺激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)、雌二醇(estradiol,E2)含量,HE染色观察下丘脑、垂体、海马等组织病理损伤情况,RT-PCR和Western blot法检测下丘脑、海马组织雌激素受体基因及蛋白的表达。结果与正常对照组比较,模型组大鼠糖水偏好度显著下降(P<0.01),强迫游泳不动时间显著增加(P<0.01),表现出明显抑郁样行为,血清GnRH、FSH、LH含量均显著升高(P<0.01或P<0.05),E2降低(P<0.01),下丘脑、垂体及海马组织受损,雌激素受体表达显著降低(P<0.01);经加味百合地黄汤干预治疗后,模型组大鼠抑郁样行为显著改善(P<0.01或P<0.05),HPO轴亢进状态及海马损伤得以缓解,下丘脑、海马雌激素受体表达均显著增加(P<0.01或P<0.05)。与阳性药物组比较,加味百合地黄汤高剂量组下丘脑雌激素受体基因表达显著增加(P<0.05)。结论加味百合地黄汤通过抑制HPO轴亢进及增加脑内雌激素受体水平,发挥抗围绝经期抑郁症作用。

加减参芪地黄汤治疗慢性肾脏病3-4期的临床研究

目的探讨加减参芪地黄汤在慢性肾病3-4期患者临床治疗中的作用。方法将2019年1月至2023年5月在苏州市中医医院住院及门诊治疗的慢性肾病3-4期患者60例,按随机数字表分组,各30例。对照组在常规治疗的基础上辅以肾衰宁片口服,治疗组在常规治疗的基础上给予加减参芪地黄汤中药治疗。检测两组患者肌酐、尿素氮、尿酸等指标,比较两组患者中医证候积分及临床疗效。结果治疗组总有效率为86.67%,显著高于对照组(77.33%)(P<0.05)。治疗组治疗后血Scr、BUN、UA显著低于对照组,差异具有统计学意义(P<0.05)。治疗组倦怠乏力、食少纳呆、腰膝酸软、肢体困重等证候积分与对照组比较均有统计学差异(P<0.01或P<0.05)。结论加减参芪地黄汤对慢性肾病3-4期患者疗效良好,可降低血肌酐、尿素氮、尿酸,改善临床症状,延缓慢性肾脏病进展。

六味地黄汤对阿霉素肾病小鼠尿蛋白及肾小球p-p38 MAPK、α-actinin-4、synaptopodin的影响

目的探讨六味地黄汤对阿霉素肾病(adriamycin nephropaghy,ADRN)小鼠尿蛋白的干预作用及可能机制。方法30只SPF级雄性C57BL/6小鼠随机选取5只为空白组,余下小鼠以单次尾静脉注射阿霉素(0.01 g·kg^(-1))复制ADRN模型,2周后将造模成功的小鼠随机分为模型组、贝那普利组及低、中、高剂量六味地黄汤组,每组5只。空白组、模型组予等体积注射用水灌胃,贝那普利组予0.0013 g·kg^(-1)贝那普利灌胃,低、中、高剂量六味地黄汤组分别予9.75、19.5、39 g·kg^(-1)六味地黄汤灌胃,每日1次,连续8周。自动生化仪检测小鼠24 h尿蛋白定量、血清白蛋白、血清肌酐、血清钾;HE染色法、电镜观察小鼠肾脏病理改变及足突形态;免疫组织化学法、Western bolt检测小鼠肾脏磷酸化p38 MAPK(phosphorylated p38 MAPK,p-p38 MAPK)、α-辅肌动蛋白-4(α-actinin-4)、突触孔蛋白(synaptopodin)的表达;RT-PCR检测小鼠肾脏α-actinin-4、synaptopodin mRNA表达。结果与空白组比较,模型组小鼠体质量、24 h尿量、血清白蛋白降低,24 h尿蛋白定量升高(P<0.05);肾小球系膜细胞增生,部分系膜区扩大、肾小管蛋白管型,间质可见炎性细胞浸润,足突广泛融合;α-actinin-4蛋白及mRNA、p-p38 MAPK蛋白表达升高(P<0.05),synaptopodin蛋白及mRNA表达降低(P<0.05)。与模型组比较,各干预组病理改变及足突融合均得到不同程度改善,贝那普利组及中、高剂量六味地黄汤组24 h尿蛋白定量减少,高剂量六味地黄汤组p-p38 MAPK蛋白表达降低(P<0.05),低、中、高剂量六味地黄汤组synaptopodin蛋白及mRNA表达均升高(P<0.05)。贝那普利组及低、中、高剂量六味地黄汤组组间比较,各指标差异无统计学意义(P>0.05)。结论六味地黄汤可能通过抑制p38 MAPK通路活化和上调α-actinin-4、synaptopodin蛋白及mRNA表达,改善肾脏病理及足突融合,减轻足细胞损伤,减少ADRN小鼠尿蛋白。

经典名方羌活胜湿汤中12种成分的LC-MS/MS定量分析

[目的]建立液相色谱-质谱联用(LC-MS/MS)分析方法用于测定羌活胜湿汤颗粒中5-O-甲基维斯阿米醇苷、紫花前胡苷、蔓荆子黄素、羌活醇、蛇床子素、二氢欧山芹醇当归酸酯、异甘草苷、甘草苷、绿原酸、异甘草素、甘草素、阿魏酸12种成分的含量。[方法]色谱柱为ACQUITY UPLC BEH C18柱(2.1 mm×100 mm,1.7μm),流动相为0.1%的甲酸水溶液和乙腈,梯度洗脱,流速为0.3 mL/min,柱温为30℃,进样量为5μL。采用电喷雾离子源(ESI)、多反应监测模式(MRM)进行正负离子同时扫描。[结果]羌活胜湿汤颗粒中12种成分在线性范围内线性关系良好,精密度、重复性、稳定性及加样回收率试验均满足要求。[结论]实验建立的含量测定方法高效、稳定、可靠,可用于羌活胜湿汤颗粒的质量控制研究。