Network pharmacology-based exploration of molecular mechanisms underlying therapeutic effects of Jianpi Huatan Quyu recipe on chronic heart failure with spleen Qi deficiency syndrome

BACKGROUND Chronic heart failure is a complex clinical syndrome.The Chinese herbal compound preparation Jianpi Huatan Quyu recipe has been used to treat chronic heart failure;however,the underlying molecular mechanism is still not clear.AIM To identify the effective active ingredients of Jianpi Huatan Quyu recipe and explore its molecular mechanism in the treatment of chronic heart failure.METHODS The effective active ingredients of eight herbs composing Jianpi Huatan Quyu recipe were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.The target genes of chronic heart failure were searched in the Genecards database.The target proteins of active ingredients were mapped to chronic heart failure target genes to obtain the common drugdisease targets,which were then used to construct a key chemical componenttarget network using Cytoscape 3.7.2 software.The protein-protein interaction network was constructed using the String database.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed through the Metascape database.Finally,our previously published relevant articles were searched to verify the results obtained via network pharmacology.RESULTS A total of 227 effective active ingredients for Jianpi Huatan Quyu recipe were identified,of which quercetin,kaempferol,7-methoxy-2-methyl isoflavone,formononetin,and isorhamnetin may be key active ingredients and involved in the therapeutic effects of TCM by acting on STAT3,MAPK3,AKT1,JUN,MAPK1,TP53,TNF,HSP90AA1,p65,MAPK8,MAPK14,IL6,EGFR,EDN1,FOS,and other proteins.The pathways identified by KEGG enrichment analysis include pathways in cancer,IL-17 signaling pathway,PI3K-Akt signaling pathway,HIF-1 signaling pathway,calcium signaling pathway,cAMP signaling pathway,NF-kappaB signaling pathway,AMPK signaling pathway,etc.Previous studies on Jianpi Huatan Quyu recipe suggested that this Chinese compound preparation can regulate the TNF-α,IL-6,MAPK,cAMP,and AMPK pathways to affect the mitochondrial structure of myocardial cells,oxidative stress,and energy metabolism,thus achieving the therapeutic effects on chronic heart failure.CONCLUSION The Chinese medicine compound preparation Jianpi Huatan Quyu recipe exerts therapeutic effects on chronic heart failure possibly by influencing the mitochondrial structure of cardiomyocytes,oxidative stress,energy metabolism,and other processes.Future studies are warranted to investigate the role of the IL-17 signaling pathway,PI3K-Akt signaling pathway,HIF-1 signaling pathway,and other pathways in mediating the therapeutic effects of Jianpi Huatan Quyu recipe on chronic heart failure.

Current concept of Spleen Stomach theory and Spleen deficiency syndrome in TCM

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Intervention effect of traditional Chinese medicine Yi Tang Kang on metabolic syndrome of spleen deficiency

Objective:To investigate effects of herbal compound Yi Tang Kang on the spleen deficiency metabolic syndrome.Methods:Forty male Wistar rats were randomly divided into two groups:the normal control group and the MS spleen deficiency syndrome group.The control group rats were fed with standard diet and water,while MS spleen deficiency syndrome group with high fat diet and low dose intraperitoneal injection of slreptozocin.which swam to the endurance limit.After 12 weeks,the MS spleen deficiency syndrome group was randomly divided into two groups,with 13 rats in each group.Flats in model group were fed with high fat diet and conlinuouly administered with daily saline,and rats in intervention group with high fat diet were trated with traditional Chinese medicines Yi Tang Kang by gavage,2 mL/200 g at the same lime every day.10 weeks later,the expression of serum proteomics was investigated through abdominal aortic puncture and separation of serum,using isotope labeling technique,high performance liquid chromatography and four bar-Orbitrap mass spectrometer.Results:After treatment with traditional Chinese medicine yitangkang,in the model group,important carboxylesterase and retinal guanylate cyclase 2 precursor were upregulated.As for intervention group,these indesxes were raised,but immunoglobulin IgG,carnitine acetyltransferase,tubulin beta-5,and Gan Lu sugar binding protein C were down-regulated.At the same time,some new biological active substances,such as protein tyrosine kinase,beta glucosidase were also found.Conclusions:Traditional Chinese medicines Yi Tang Kang could regulate glucose and lipid metabolism in rats with spleen deficiency syndrome.

Study on the Expression of Organic Anion Transporting Polypeptide (oatp2a1) in Rat with Spleen Deficiency Syndrome and the Exploration of Clinical Significance

Objective: to explore the mechanism of transportation and transformation of dampness by the way of the expression of organic anion transporting polypeptide (oatp) superfamily member 2a1 (oatp2a1) mRNA in rat with spleen deficiency syndrome and the significance in transportation and transformation of dampness. Methods: 32 wistar male rats were divided randomly into four groups: normal group (n = 6), normal + AA group (n = 6), spleen deficiency group (n = 10), Spleen deficiency + AA group (n = 10). After reserpine-induced spleen deficiency model was made, intragastric administration of aristolochic acid (AA) was adopted for three days, the expression of oatp2a1 mRNA were detected in the tissues of lung, liver, kidney, stomach, small intestine and large intestine in four groups by using Fluorescent Quantitative-Polymerase Chain Reaction (FQ-PCR). Results: the expression of oatp2a1 mRNA in above six tissues could be detected. The ex-pression of oatp2a1 mRNA in liver tissue of rat with spleen deficiency syndrome was up-regulated compared to normal group (P = 0.035, P < 0.05), the expression of oatp2a1 mRNA in small intestinal tissue of rat with spleen deficiency syndrome was down-regulated compared to normal group (P = 0.004, P < 0.01), the expression of oatp2a1 in intestinal tissue in normal + AA group is down-regulated compared to normal group (P = 0.032, P < 0.05). Conclusions: oatp2a1 might be one of the material basis involved in transportation and transformation of dampness. The changes of expression of oatp2a1 mRNA in small intestine, liver tissue suggests that small intestine, liver might play an important role in the transportation and transformation of dampness in the state of spleen deficiency. We further concluded that the function of spleen’s governing transportation and transformation of dampness was not only including the function of the gastrointestinal, but also part of the liver function in some degree, which needs to be further studied.

Biological basis of “depression with liver-qi stagnation and spleen deficiency syndrome”: A digital gene expression profiling study

Objective:To investigate the biological basis of“depression with liver-qi stagnation and spleen deficiency syndrome”.Methods:A digital gene expression profiling method was conducted to explore global changes in the mRNA transcriptome in a rat model of depression with liver-qi stagnation and spleen deficiency syndrome.Real-time quantitative polymerase chain reaction(q-PCR)was performed to verify the five genes most interest based on the Kyoto Encyclopedia of Genes and Genome(KEGG)analysis.Sini San,which disperses stagnated liver qi and strengthens the spleen,was administered to the model rats to observe whether it could reverse these genetic changes in the liver.Results:Forty-six differentially expressed genes were identified.Three of the five genes of most interestdHnf4a,Hnf4g and Cyp1a1dbased on KEGG analysis,were confirmed by realtime q-PCR.Sini San reduced the gene expression changes of Hnf4a,Hnf4g and Cyp1a1 in the rat model.Conclusions:Hnf4a,Hnf4g and Cyp1a1 are involved in“depression with liver-qi stagnation and spleen deficiency syndrome”.These findings indicate that depressed rats with liver-qi stagnation and spleen deficiency syndrome are at risk of liver diseases.Furthermore,our results will inform exploration of the etiology of depression and help in the development of effective therapeutic strategies.

Rules of Ren Shunping's prescription for trearing Spleen Deficiency and Qi Stagnation Syndrome of Functional Dyspepsia Based on Data Mining

Objective:To explore the medication rules of Professor Ren Shunping treating spleen deficiency and qi stagnation syndrome of functional dyspepsia,so as to provide reference for clinical prevention and development of new drugs.Methods:The 144 Chinese herbal formulas considered as active prescribed by professor Ren Shunping for the first time treating the patients with spleen deficiency and qi stagnation syndrome of functional dyspepsia were collected,entered into TCM inheritance auxiliary platform(V2.5),and then mined by some data mining methods of the software,such as the association rules and unsupervised entropy hierarchical clustering.Results:This study involved 144 Chinese herbal formulas,including a total of 56 Chinese herbal medicines.Of these Chinese herbal medicines,the herbs of cold and warm nature,and pungent,bitter and sweet in flavors were used in a quite high frequency,all of which act on spleen,stomach,lungs,large intestine,liver and gallbladder meridians.One core Chinese herbal formula was excavated,composed of patchouli,perilla stem,radix bupleuri,scutellaria,ginger pinellia,codonopsispilosula,magnolia bark,tangerine peel,large-headed atractylodes,immature bitter orange,wood incense,fructus amomi,inula britannica chinensis,ginger,medicated leaven,fried rice and fried malt based on the association rules.And a total of 17 groups of core used drugs and 2 groups of new Chinese herbal formulas undiscovered were mined based on unsupervised entropy hierarchical clustering.Conclusion:In treating spleen deficiency and qi stagnation syndrome of functional dyspepsia,professor Ren is good at treating spleen deficiency and qi stagnation syndrome of functional dyspepsia from the perspective of"tong jiang",using the"five-in-one"strategy of strengthening the spleen,evacuating the liver,coming down qi from the stomach,dispersing lung,and bowel-reflexing to treat patients as a whole.And the herbal medicines were light in dose,lovely and mild in nature for the formulars.At the same time,he paid attention to the combination of syndromes and symptoms,and flexible arbitration.

Study on the Characteristics of Gut Microbiota in Chronic Hepatitis B(CHB)Patients with Damp Heat Syndrome and Liver Depression and Spleen Deficiency Syndrome

Objective:This study was designed to explore the characteristics of gut microbiota in CHB patients with two most common traditional Chinese medicine(TCM)Syndromes—damp heat syndrome and liver depression and spleen deficiency syndrome.Methods:According to the diagnostic criteria of Western medicine,TCM and screening exclusion criteria,65 cases of CHB with damp heat syndrome and 28 cases of CHB with liver depression and spleen deficiency syndrome were finally included in the study.All the basic information was gathered and the fresh fecal samples were collected for 16S rDNA sequencing.16S rDNA of gut microbiota was sequenced using Illumina hiseq 2,500 high-throughput sequencing platform.Based on the optimized sequence,Operational Taxonomic Units(OTU)clustering analysis and taxonomic annotation were carried out.Results:The difference in relative abundance of gut microbiota was significant between damp heat syndrome and liver depression and spleen deficiency syndrome in CHB patients.Cyanobacteria was only found in damp heat syndrome.The relative abundance of Erysipelotrichia and Subdoligranulum were higher in liver depression and spleen deficiency syndrome,while the relative abundance of Rhodospirillales,Alphaproteobacteria and Lachnospira were higher in the damp heat syndrome.LDA Effect Size(LEfSe)analysis showed that Lachnospira,Olsenella and Subdoligranulum had significant difference in species among the two TCM syndromes.Conclusion:The different characteristics of gut microbiota in the two TCM syndromes of CHB patients may play an important role in syndrome formation of TCM,which provides a new field of vision for the accurate diagnosis and treatment of TCM.

Efficacy of Pidan Jianqing decoction in treatment of type 2 diabetes with spleen deficiency and damp-heat syndrome

Introduction:This study aimed to assess the clinical efficacy of Pidan Jianqing decoction in the treatment of type 2 diabetes.Methods:A total of 72 patients with type 2 diabetes differentiated as spleen deficiency with damp-heat syndrome were randomly assigned to a treatment group(n=35)or control group(n=32).Patients in the control group received diet and exercise guidance and medication in the form of 0.5 g metformin hydrochloride tablets,while patients in the treatment group received Pidan Jianqing decoction in addition to the treatment given to the control group.Efficacy was evaluated after 8 weeks of treatment.Results:Efficacy was 91.4%in the treatment group versus 68.8%in the control group,which was a statistically significant difference(P<0.05).Significant group differences were also noted in the number of patients with improvement in symptoms such as dry mouth and polydipsia,frequency of urination and polyuria,abdominal distention and loss of appetite,fatigue and weakness,and sticky stools(all P<0.05),with greater improvement in the treatment group compared to the control group.The fasting blood glucose(FBG)and 2 h postprandial blood glucose levels significantly improved in both groups after treatment(P<0.05);however,there was no significant difference in glycated hemoglobin levels before and after treatment in both groups.Conclusion:Pidan Jianqing decoction can lower blood glucose levels and improve symptoms in patients with type 2 diabetes.

Clinical study on fire needle acupuncture treating irregular menstruation with spleen deficiency syndrome

Objective:To observe the clinical efficacy of fire needle acupuncture in treating irregular menstruation with spleen deficiency syndrome.Methods:60 patients with spleen deficiency-irregular menstruation were randomly divided into treatment group and control group of 30 each.In the treatment group,fire needle acupuncture was used to insert into Guanyuan,Qihai,Pishu,Sanyinjiao,Zusanli and Taibai points,while the filiform needle was used to needle the corresponding acupoints in the control group.After 8 weeks of treatment,menstrual cycle,menstrual period,menstrual volume,irregular menstrual symptom score and spleen deficiency symptom score were recorded.Results:Both treatments could effectively improve the menstrual cycle,menstrual period and menstrual volume,but there was no statistical significance between the two groups(P>0.05).Both treatments were effective in regulating the symptoms of menstruation,and fire needle acupuncture was better than filiform needle(P<0.05).Both treatments relieved the symptoms of spleen deficiency,and fire needle acupuncture was better than filiform needle(P<0.05).In terms of overall curative effect,the curative effect of fire needle acupuncture therapy was significantly better than filiform needle therapy,and the difference was statistically significant(P<0.05).Conclusion:Fire needle acupuncture therapy could effectively regulate menstrual volume of patients with irregular menstruation of spleen deficiency syndrome,and relieve the symptoms of traditional Chinese medicine.

Abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type

Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in the method were from the Physical Examination Center of Anhui Provincial Hospital of Traditional Chinese Medicine,and the 60 rheumatoid arthritis patients included were from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine.The disease activity score of patients with rheumatoid arthritis was evaluated using VAS score and DAS28 score,joint symptoms and spleen deficiency syndrome score were evaluated using graded quantification method,immune inflammation indicators were detected using relevant instruments,inflammatory cytokines were detected using ELISA method,macrophage markers were detected using FCM method,and pathway gene expression was detected using RT-qPCR;Evaluate the predictive effect of circ-CBLB and miR-486-5p on disease activity in rheumatoid arthritis using ROC curves.Results:(1)miR-486-5p,CD14^(+)CD86^(+),ESR,CRP,RF,Anti CCP Ab,IL-6,TNF in patients with rheumatoid arthritis-αThe levels of circ-CBLB,CD14^(+)CD163^(+),IL-4,and IL-10 were significantly higher than those of healthy individuals;(2)The expression level of circ-CBLB in patients with rheumatoid arthritis is positively correlated with CD14^(+)CD163^(+),and negatively correlated with miR-486-5p and CD14^(+)CD86^(+);The expression level of miR-486-5p is negatively correlated with CD14^(+)CD163^(+)and positively correlated with CD14^(+)CD86^(+);There is a negative correlation between CD14^(+)CD86^(+)and CD14^(+)CD163^(+);ESR is negatively correlated with circ-CBLB,and positively correlated with miR-486-5p,CD14^(+)CD86^(+),CRP;CRP is negatively correlated with circ-CBLB,CD14^(+)CD163^(+),and positively correlated with CD14^(+)CD86^(+),ESR;(3)The expression level of circ-CBLB in patients with rheumatoid arthritis is negatively correlated with joint tenderness,morning stiffness,lack of qi and lazy speech,and postprandial abdominal distension score;The expression level of miR-486-5p is positively correlated with the scores of joint tenderness and decreased appetite.(4)The ROC curve shows that in terms of circ-CBLB,ESR,CRP,VAS,and DAS28 AUC are 0.662(P=0.032),0.658(P=0.035),0.516(P=0.830),and 0.791(P=0.000),respectively.In terms of miR-486-5p,ESR,CRP,VAS,and DAS28 AUC were 0.566(P=0.385),0.511(P=0.883),0.592(P=0.223),and 0.727(P=0.003),respectively.Conclusion:The abnormal expression of circ CBLB and miR-486-5p in peripheral blood mononuclear cell of patients with rheumatoid arthritis of spleen deficiency and dampness excess type is related to inflammatory polarization markers,immune inflammation,disease activity,joint symptoms and spleen deficiency syndrome of rheumatoid arthritis,and the low expression of circ CBLB and high expression of miR-486-5p have certain predictive value for disease activity of rheumatoid arthritis.

Exosomes promote the invasion and metastasis of hepatocellular carcinoma cells via an integrin-dependent manner in the spleen-deficient state

Background:To investigate the detailed mechanism underlying the pro-metastatic effect of spleen deficiency(SD)syndrome on hepatocellular carcinoma(HCC).Methods:In the present study,our model was established based on an HCC mouse model induced by diethylnitrosamine using reserpine to induce SD.Exosomes were isolated and purified from mouse plasma samples using an exosome isolation kit.Subsequently,we verified the pro-metastatic effects of exosomes from the HCC mice with SD on HCC cells by transwell assays,wound healing assays,phalloidin staining in vitro,and lung metastasis assay of mice in vivo.Finally,we further explored the detailed mechanism underlying the pro-metastatic effect of exosomes from the HCC mice with SD on HCC cells.Results:We found that SD promoted the malignant progression of HCC in mice.Exosomes from HCC mice with SD enhanced the invasion and metastasis of HCC cells in vitro and in vivo.Mechanistically,upregulation of integrinα1,integrinβ1,and integrinβ5 seemed to play a key role in mediating the pro-metastatic effect of exosomes isolated from the HCC mice with SD,which was largely abrogated upon co-treatment with a broad-spectrum integrin inhibitor.Conclusion:Our findings demonstrated that exosomes promote the invasion and metastasis of HCC cells via an integrin-dependent manner in the spleen-deficient state that would contribute to our better understanding of the role of SD in HCC progression in traditional Chinese medicine,and thus management of the disease.

The prescription rule analysis of the New Theory on Spleen Dampness Syndrome by Academician TONG Xiaolin

Objective To analyze the basic characteristics,drug features,prescription rules,and drug-symptom relationships of patients in the splenic deficiency and impairment stage,by data mining of medical records under the New Theory on Spleen Dampness Syndrome(Pi Dan Xin Lun,《脾瘅新论》).Methods Medical records listed in the“New Theory on Spleen Dampness Syndrome-Under-standing and Treatment of Metabolic Syndrome from the Perspective of Traditional Chinese Medicine”,and which were diagnosed with the spleen dampness syndrome at the splenic de-ficiency and impairment stage,during January 2004 and December 2016 were selected.These patients’data,including basic information,clinical symptoms,laboratory examination res-ults,traditional Chinese medicine(TCM)and western medicine diagnoses,treatment meth-ods,prescriptions,etc.,were collected.The collected data were subsequently compiled into a medical record database using the Epidata 3.1 data management software,followed by the use of Apriori algorithm provided in the SPSS Modeler 14.2 statistical software to investigate the association rules between drug-drug,drug-symptom,and drug-western medicine indices.Results(i)A total of 51 medical records were included,involving 17 types of syndromes.Among them,the top three with frequency≥3 included“Phlegm and blood stasis,and thoracic obstruction”“Deficiency-weakness of the spleen Qi,and static blood blocking collat-erals”,and“Deficiency-weakness of the spleen Qi,and static blood blocking collaterals”.Al-ternatively,of the 14 treatment methods,the top three treatments with frequency of≥3 in-cluded“Activating Yang and eliminating turbidity,and removing phlegm and dredging chan-nel blockage”“Strengthening the spleen and benefiting Qi,and eliminating phlegm to activ-ate the channels”,and“Warming Yang and benefiting Qi,and expelling cold to remove ob-structions”.Among the 15 prescriptions,the top three used with frequency≥3 included Huangqi Guizhi Wuwu Tang(黄芪桂枝五物汤),Gualou Xiebai Banxia Tang(瓜蒌薤白半夏汤),and Ganjiang Huangqin Huanglian Renshen Tang(干姜黄芩黄连人参汤).Lastly,of the 83 drugs used for a total of 476 times,those with frequency≥15 included Huanglian(Coptid-is Rhizoma),Huangqi(Astragali Radix),Jiudahuang(Wine-processed Rhei Radix et Rhizoma),Jixueteng(Spatholobi Caulis),Shengjiang(Zingiberis Rhizoma Recens),Huangqin(Scutellariae Radix),and Guizhi(Cinnamomi Ramulus).(ii)For the drug-drug associations,under the criteria of support≥15%and confidence=100%,seven second-order association rules,seven third-order rules,and six fourth-order roles were identified.The top-ranking rule of each was“Huangqin(Scutellariae Radix)→Huanglian(Coptidis Rhizoma)”“Ganjiang(Zingiberis Rhizoma)+Huangqin(Scutellariae Radix)→Huanglian(Coptidis Rhizoma)”,and“Baishao(Paeoniae Radix Alba)+Guizhi(Cinnamomi Ramulus)+Jixueteng(Spatho-lobi Caulis)→Huangqin(Scutellariae Radix)”,respectively.Alternatively,the drug-symptom associations were analyzed under the criteria of support≥5%and confidence=100%,which derived eight second-order association rules,31 third-order rules,and 30 fourth-order rules.The top-ranking association rule of each order was“Huangqi(Astragali Radix)→Limb ed-ema”“Guizhi(Cinnamomi Ramulus)+Jixueteng(Spatholobi Caulis)→Limb numbness and pain”,and“Guizhi(Cinnamomi Ramulus)+Jixueteng(Spatholobi Caulis)+Huangqi(As-tragali Radix)→Limb numbness and pain”,respectively.Similarly,the drug-western medi-cine index associations were investigated under the criteria of support≥5%and confidence=100%,and five second-order association rules,16 third-order rules,and 16 fourth-order rules were identified.In this category,the top-ranking association rule of each order was“Qinpi(Fraxini Cortex)→Uric acid”“Huanglian(Coptidis Rhizoma)+Ganjiang(Zingiberis Rhizoma)→Glycated hemoglobin”,and“Huanglian(Coptidis Rhizoma)+Ganjiang(Zing-iberis Rhizoma)+Huangqin(Scutellariae Radix)→Glycated hemoglobin”,respectively.Conclusion Through association rule mining,this study objectively and quantitatively demonstrated the drug-drug,drug-symptom,and drug-physicochemical index associations of patients with the spleen dampness syndrome at the splenic deficiency and impairment stage treated by Academician TONG Xiaolin.The results indicated that treatment for these patients adopted the“state-target”syndrome differentiation method.The drug combination was characterized by“small prescriptions”,targeting both the patient’s symptoms and signs(syndrome target)and western medicine indices(treatment target).This study could provide references for future research on the academic thoughts and medical experience of Academi-cian TONG Xiaolin.

EFFECTS OF MEDICINAL MOXIBUSTION ON SERUM GASTRIN,URINARY D-XYLOSE EXCRETORY RATE AND T CELL SUBGROUPS IN SPLEEN DEFICIENCY PATIENTS

The medicinal moxa stick made by moxa wool combining with Chinese medicines of supporting the body resistance and strengthening the spleen was used to treat 74 cases of spleen deficiency, the other 41 patients as control were treated by pure moxa stick. After treatment, the serum gastrin, urinary D - xylose excretory rate and T cell subgroups in patients with spleen deficiency have been greatly improved as compared with those before treatment in two groups (P< 0.01 or 0.05). Comparing the changes of the indexes between treatment group and control group, there were significant(P<0.05), or markedly significant(P<0.01) differences. It indicated that the medicinal moxibustion could improve the secreting and absorbing functions of digestive tract and the cellular immune function in patients with spleen deficiency.

Screening the effective components in treating dampness stagnancy due to spleen deficiency syndrome and elucidating the potential mechanism of Poria water extract

Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer.However,the specific effective components and the potential mechanism of Poria remain largely unknown.To identify the effective components and the mechanism of Poria water extract(PWE)to treat dampness stagnancy due to spleen deficiency syndrome(DSSD),a rat model of DSSD was established through weight-loaded forced swimming,intragastric ice-water stimulation,humid living environment,and alternate-day fasting for 21 days.After 14 days of treatment with PWE,the results indicated that PWE increased fecal moisture percentage,urine output,D-xylose level and weight;amylase,albumin,and total protein levels;and the swimming time of rats with DSSD to different extents.Eleven highly related components were screened out using the spectrum-effect relationship and LC-MS.Mechanistic studies revealed that PWE significantly increased the expression of serum motilin(MTL),gastrin(GAS),ADCY5/6,p-PKAα/β/γcat,and phosphorylated cAMP-response element binding protein in the stomach,and AQP3 expression in the colon.Moreover,it decreased the levels of serum ADH,the expression of AQP3 and AQP4 in the stomach,AQP1 and AQP3 in the duodenum,and AQP4 in the colon.PWE induced diuresis to drain dampness in rats with DSSD.Eleven main effective components were identified in PWE.They exerted therapeutic effect by regulating the AC-cAMP-AQP signaling pathway in the stomach,MTL and GAS levels in the serum,AQP1 and AQP3 expression in the duodenum,and AQP3 and AQP4 expression in the colon.

Efficacy and Safety of Jianpi Jieyu Decoction for Patients with Mild-to-Moderate Depression of Xin(Heart)-Pi(Spleen)Deficiency Syndrome:A Multi-centre Randomized Controlled Study

Objective:To evaluate the efficacy and safety of Jianpi Jieyu Decoction(JJD)for treating patients with mild-to-moderate depression of Xin(Heart)-Pi(Spleen)deficiency(XPD)syndrome.Methods:In this multi-center,randomized,controlled study,140 patients with mild-to-moderate depression of XPD syndrome were included from Xiyuan Hospital of China Academy of Chinese Medical Sciences and Botou Hospital of Traditional Chinese Medicine from December 2017 to December 2019.They were randomly divided into JJD group and paroxetine group by using a random number table,with 70 cases in each group.The patients in the JJD group were given JJD one dose per day(twice daily at morning and evening,100 m L each time),and the patients in the paroxetine group were given paroxetine(10 mg/d in week 1;20 mg/d in weeks 2-6),both orally administration for a total of 6 weeks.The primary outcome was the change of 17-item Hamilton Depression Rating Scale(HAMD-17)score at week 6 from baseline.The secondary outcomes included the Hamilton Anxiety Scale(HAMA)score,Traditional Chinese Medicine Symptom Scale(TCMSS),and Clinlcal Global Impression(CGI)scores at the 2nd,4th,and 6th weekends of treatment,HAMD-17 response(defined as a reduction in score of>50%)and HAMD-17 remission(defined as a score of 7)at the end of the 6th week of treatment.Adverse events(AEs)were also recorded.Results:From baseline to week 6,the HAMD-17 scores decreased 10.2±4.0 and 9.1±4.9 points in the JJD and paroxetine groups,respectively(P=0.689).The HAMD-17 response occurred in 60%of patients in the JJD group and in 50%of those in the paroxetine group(P=0.292);HAMD-17 remission occurred in 45.7%and 30%of patients,respectively(P=0.128).The differences of CGI scores at the 6th week were not statistically significant(P>0.05).There were significant differences in HAMD-17 scores between the two groups at 2nd and 4th week(P=0.001 and P=0.014).The HAMA scores declined 8.1±3.0 and 6.9±4.3 points from baseline to week 6 in the JJD and paroxetine groups,respectively(P=0.905 between groups).At 4th week of treatment,there was a significant difference in HAMA between the two groups(P=0.037).TCMSS decreased 11.4±5.1,and 10.1±6.8 points in the JJD and paroxetine groups,respectively(P=0.080 between groups).At the 6th week,the incidence of AEs in the JJD group was significantly lower than that in the paroxetine group(7.14%vs.22.86%,P<0.05).Conclusion:Compared with paroxetine,JJD was associated with a significantly lower incidence of AEs in patients with mild-tomoderate depression of XPD syndrome,with no difference in efficacy at 6 weeks.(Trial registration No.Chi CTR2000040922).

Study on the Resolving Phlegm Effect of D-Limonene in Mice with Spleen Deficiency and Phlegm-Dampness Syndrome

Objective:According to Traditional Chinese Medicine theory,spleen deficiency and phlegm-dampness syndrome(SDPDS)are caused by abnormal water metabolism in the body because of spleen dysfunction.The purpose of this article was to evaluate the efficacy of D-limonene(DL)in resolving phlegm in mice with SDPDS from the perspective of regulating the level of aquaporin 3(AQP3).Methods:The model of SDPDS was induced in mice using the multifactor modeling method,which combines internal and external dampness.An artificial climate box was used to create a humid environment,whereas the irregular diet was caused by different feeding methods on odd-even days.The mice were divided into blank control,model group,DL low-dose,DL high-dose,and positive groups.The mice were modeled and treated for 7 day.Levels of gastrin and amylase(AMS)in the serum,mucus secretion in the trachea,and AQP3 in the tissue near the gastric cardia.Results:DL significantly reduced mucus secretion in the trachea(P<0.001).It also increased the level of AMS in the serum(P<0.01)and decreased the level of AQP3 in the gastric tissue(P<0.01).Conclusions:Mice with SDPDS exhibited disturbed water metabolism and significantly increased AQP3 levels.DL can restore the levels of AQP3 and plays an important role in resolving phlegm.This study may also help expand the efficacy of natural drugs containing DL and improve the utilization of natural drug resources.

Pharmacokinetics of traditional Chinese syndrome and recipe:a hypothesis and its verification(Ⅰ)

AIM To propose a hypothesis defining theabsorption,distribution,metabolism andelimination of traditional Chinese recipe（TCR）-component in blood of healthy subjects andpatients,and estimate its correctness.METHODS The pharmacokinetics（PK）of samedose of drug was studied in the animal model oftraditional Chinese syndrome（S）and healthyanimals.The classification,terminology,concept and significance of the hypothesis wereset forth with evidence provided in the presentstudy.The hypotheses consisted of traditionalChinese syndrome PK（S-PK）and traditionalChinese recipe PK（R-PK）.Firstly,the observedtetramethylpyrazine（TMP）PK in healthy,chronically reserpinized rats（rat model ofspleen deficiency syndrome,RMSDS）andRMSDS treated with Sijunzi decoction（SJZD）forconfirmation were used to verify S-PK; secondly,the ferulic acid（FA）PK in healthy andhigh molecular weight dextran（HMWD）-inducedrabbit model with blood stasis syndrome（RDBSS）was also used to verify S-PK;andlastly,TMP PK parameters in serum of healthyrats after orally taken Ligusticum wallichii（LW）,LW and Salvia miltiorrhiza（LW&SM）decoctions were compared to verify R-PK.RESULTS The apparent first-order absorption[Ka,（13.61±2.56）h-1,area under the blooddrug concentration-time curve[AUC,（24.88±9.76）μg.h-1mL-1],maximum drug concentration[Cmax,（4.82±1.23）μg·mL-1]of serum TMP inRMSDS were increased markedly（P【0.05）compared with those[Ka=（5.41±1.91）h-1,AUC=（5.20±2.57）μg·h-1·mL-1,Cmax=（2.33±1.77）μg·mL-1]of healthy rats（HR）.Theapparent first-order rate constant for α and βdistribution phase[α=（0.38±0.09）h-1,β=（0.06±0.03）h-1,the apparent first-orderintercompartmental transfer rate constants[K10=（0.24±0.07）h-1,K12=（0.11±0.02）h-1,K21=（0.11±0.02）h-1]of serum TMP in RMSDS weredecreased significantly（P【0.01）comparedwith those[K10=（0.88±0.20）h-1,K12=（1.45±0.47）h-1,K21=（0.72±0.22）h-1]of HR.However,no apparent differences occurredbetween HR and RMSDS treated with SJZD.Theserum FA concentration and its AUC[（5.6690±2.3541）μg·h-1·mL-1] in RMBSS were also higherthan those[AUC=（2.7566±0.8232）μg·h-1·mL-1]of healthy rabbits（P【0.05）.The Ka（11.51±2.82）h-1,AUC（0.84±0.17）μg·h-1·mL-1of LW & SM-derived TMP in serum weremuch lower（P【0.05）than those[Ka=（19.58±4.14）h-1,AUC=（1.27±0.26）μg·h-1·mL-1]ofLW-derived TMP in serum after oral decoctions.CONCLUSION The SDS and blood stasissyndrome state could affect significantly thepharmacokinetic parameters of drugs and theabnormal SDS pharmacokinetic parameters couldbe normalized by SJZD.The combination ofChinese medicine in TCR could reciprocallyaffect the pharmacokinetic parameters of othercomponents absorbed into the systemiccirculation.These results support the S-and R-PK hypothesis.

Study of establishing disease-syndrome combined with animal model for immune thrombocytopenic purpura without additional conditions

Objective:To explore the feasibility of establishing the disease-syndrome combined animal model for immune thrombocytopenic purpura(ITP)without additional conditions.Methods:Three batches of data related to the ITP model mice obtained by replication at different time were analyzed,and whether the APS-injected model mice replicated through the passive immune modeling method could simulate the pathogenesis and clinical characteristics of human ITP was evaluated according to the differentiation criteria for diseasesyndrome combined model.Results:The APS-injected replicated ITP model mice possessed the following traits:(1)Compared with the normal group,the platelet count was significantly decreased,and coagulation time was significantly increased in the model group(P<.01).(2)Compared with the normal group,the medullary thrombocytogenous megakaryocytes were significantly decreased(P<.05,.01,.001).(3)The APS-injected sites and other parts of the model mice had spontaneous hemorrhage.(4)Behavioral changing signs were observed 1 week after the modeling(i.e.low activity,delayed activity,poor appetite,skin petechia/hemorrhage and spontaneous hemorrhage at the injected sites or other parts),and were getting more and more severe.Conclusion:According to the syndrome differentiation criteria for disease-syndrome combined model of ITP,the APS-injected animal model of ITP replicated through the passive immune modeling method without additional conditions possesses the characteristics of disease-syndrome combined model.It provides an ideal tool for the development of traditional Chinese medicine pharmacology experiment.

The mechanism of Astragalus membranaceus in treating peritoneal fibrosis by intervening the key syndrome and pathology based on Q-marker theory

Objective:To evaluate the mechanism of Astragalus membranaceus(AM)by intervening peritoneal mesothelial cells(PMCs),epithelial-mesenchymal transition(EMT),and spleen deficiency syndrome(SDS)in peritoneal fibrosis(PF),we base on employing the strategy of Q-marker theory combination network pharmacology method.Methods:First,we obtained the Q-markers of AM by searching the relevant literature and its pharmacological information was collected based on SwissADME.The SwissTargetPrediction and pharmmaper were employed to predict its potential target.Secondly,GeneCards,DisGeNET,and OMIM databases were employed to search the related targets of EMT,SDS,and PF.VENNY2.1 tool was employed to obtain the intersection targets of AM and the three;then the“AM potential target-SDS-EMT-PF”Venn diagram was constructed.The common targets of AM,EMT and SDS were uploaded to the STRING database and obtained the PPI protein interaction network map.Cytoscape 3.7.2 was employed to evaluate the core target of PPI network.PATHER and Metascape databases were used to analyze protein type,GO biological process,and KEGG pathway.Finally,a network diagram of the“TCM-component-disease target-pathway”was drawn.Results:A total of 10 AM Q-makers were screened out,corresponding to 335 targets of AM,2,728 targets of SDS,373 of EMT,and 612 PF targets were found.Among them,there are 155 common AM targets related to SDS and EMT.Key targets such as ALB,AKT1,VEGFA,TNF,EGFR,CASP3,SRC,STAT3,HSP90AA1,and ESR1 were obtained.The core drug include quercetin,astragalosideIII,Calycosin-7-O-beta-D-glucoside,astragalosideIV,etc.The types of PPI proteins include protein modification enzymes,metabolite transferases,transmembrane signal receptors,etc.Biological processes include the regulation of kinase activity,the positive regulation of transferase activity,and the regulation of kinase activity.The key pathways may include PI3K-Akt signaling pathways,the non-smad pathway of the TGF-βsignaling pathway,and the AGE-RAGE signaling pathway.Conclusion:AM could prevent and treat PF through a multi-component multi-target-multi-path mechanism.Astragalus saponins may be the main component types of AM intervening EMT pathology by strengthening the spleen and nourishing Qi.AstragalosideIV and astragalosideIII may be the constituents that can invigorate the spleen and replenish Qi.The results of this study contribute to a more comprehensive understanding of the critical components and mechanisms of AM by intervening SDS and EMT in the treatment of PF.

基于靶向代谢组学研究苍术麸炒前后干预脾虚大鼠氨基酸水平变化的影响

目的研究采用靶向代谢组学方法,观察苍术麸炒前后对脾虚大鼠血清氨基酸水平的变化,探讨苍术炮制机制。方法72只大鼠随机分成正常组、模型组、低剂量生苍术组(生低组)、中剂量生苍术组(生中组)、高剂量生苍术组(生高组)、低剂量麸炒苍术组(麸炒低组)、中剂量麸炒苍术组(麸炒中组)和高剂量麸炒苍术组(麸炒高组),共8组,每组9只。除正常组外,模型组通过过度疲劳、苦寒泻下和饮食不节法造模。对各组大鼠采用酶联免疫吸附测定(Enzyme Linked Immunosorbent Assay,ELISA)方法测定颌下腺中水通道蛋白3(Aquaporin-3,AQP-3)、水通道蛋白4(AQP-4)、水通道蛋白5(AQP-5)、水通道蛋白8(AQP-8)的含量,结肠中水通道蛋白1(AQP-1)、水通道蛋白2(AQP-2)、水通道蛋白9(AQP-9)、紧密连接蛋白1(zonula occludes protein-1,ZO-1)的含量。对生中组和麸炒中组采用超高效液相色谱-三重四极杆质谱(Ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry,UH-PLC-MS/MS)检测血清中24种氨基酸水平的变化。结果与正常组比较,模型组中颌下腺AQP-3、AQP-4、AQP-5、AQP-8的含量显著降低(P<0.05);模型组中结肠AQP-1和AQP-2的含量显著升高(P<0.05),AQP-9和ZO-1的含量显著降低(P<0.05)。生苍术和麸炒苍术对上述指标均有调节作用。经进一步同等剂量的生苍术和麸炒苍术相比,麸炒中组使上述指标含量升高更明显。与正常组比较,模型组血清中L-缬氨酸等4种物质的水平显著升高(P<0.05),L-酪氨酸等11种物质的水平均显著降低(P<0.05)。经进一步比较,麸炒苍术对上述指标调节效果优于生苍术。靶向代谢通路分析表明,这些差异代谢物主要与精氨酸和脯氨酸代谢、氨酰tRNA生物合成、甘氨酸、丝氨酸和苏氨酸代谢、乙醛酸和二羧酸代谢通路有关。结论苍术炮制后的增效机制可能与对脾虚大鼠血清中氨基酸调节有关。