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.

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.

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.

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.

Appraisal of treatment outcomes in integrative medicine using metabonomics:Taking non-alcoholic fatty liver disease with spleen deficiency syndrome as an example

Objective:Appraisal of treatment outcomes in integrative medicine is a challenge due to a gap between the concepts of Western medicine(WM)disease and traditional Chinese medicine(TCM)syndrome.This study presents an approach for the appraisal of integrative medicine that is based on targeted metabolomics.We use non-alcoholic fatty liver disease with spleen deficiency syndrome as a test case.Methods:A patient-reported outcome(PRO)scale was developed based on literature review,Delphi consensus survey,and reliability and validity test,to quantitatively evaluate spleen deficiency syndrome.Then,a metabonomic foundation for the treatment of non-alcoholic fatty liver disease with spleen deficiency syndrome was identified via a longitudinal interventional trial and targeted metabolomics.Finally,an integrated appraisal model was established by identifying metabolites that responded in the treatment of WM disease and TCM syndrome as positive outcomes and using other aspects of the metabonomic foundation as independent variables.Results:Ten symptoms and signs were included in the spleen deficiency PRO scale.The internal reliability,content validity,discriminative validity and structural validity of the scale were all qualified.Based on treatment responses to treatments for WM disease(homeostasis model assessment of insulin resistance)or TCM syndrome(spleen deficiency PRO scale score)from a previous randomized controlled trial,two cohorts comprised of 30 participants each were established for targeted metabolomics detection.Twenty-five metabolites were found to be involved in successful treatment outcomes to both WM and TCM,following quantitative comparison and multivariate analysis.Finally,the model of the integrated appraisal system was exploratively established using binary logistic regression;it included 9 core metabolites and had the prediction probability of 83.3%.Conclusion:This study presented a new and comprehensive research route for integrative appraisal of treatment outcomes for WM disease and TCM syndrome.Critical research techniques used in this research included the development of a TCM syndrome assessment tool,a longitudinal interventional trial with verified TCM treatment,identification of homogeneous metabolites,and statistical modeling.

Study on Mitochondrial Ultrastructure, Traco Elementsand Their Correlative Factore in Cells ot Gastric Mucosa ot theGastropathic Patienta with Spleen Deficiency Syndrome

Atfer examining 88 gastropathic patieiits with Spleen deficiency syndrome by iJsing transmis-sion electron microscope, X-ray energy disperse analysis system, histochemical staining and radioimmunomethods, the authors found that the gastric mucosa cyclic adenosine monophosphate, superoxide dismutaselevel, quantity of mitochondria and its crista, the ratio of diameter between ventricle and cavity of mitochon-dria aiid the conteiit ot zinc (Zn) , copper (Cu) of mitochondria were decreasing to certain extent which tendsto get lower and lower with different groups in the order of health coritrol group, Spleen Qi deficiency groupaiid SPIeen deficiency with Qi stagnation group; chronic superficial gastritis group, chronic atrophic gastritisgroup and gastric cancor group , complete small intestinal metaplasia (IM) group, incomplete small iM group,complete colonic iM groiJp and incomplete colonic iM group (P< 0 . 05￣0 . 001 ) . While tlie degeiieratiori rateof mitochondria, Cu/Zn ratio, metaplasia rate of gastric mucosa, rate of incomplete colonic IM and content ofIipid peroxide were increasing in the above order (P < 0 . 05 ￣0 . 001 ) . It is suggested that tlie comprehensiveeffect ot the degeneration of mitochoridria and the quantitative changes of its correlative factors is the phys-iopathologic base for indecing SPIeen deficiency disease, gastric mucosa iM and canceration.

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.

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.

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.

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.

Potential Mechanism and Application of Strengthening the Spleen, Clearing Heat and Dampness in Treatment of Gastroesophageal Reflux Disease

Gastroesophageal reflux disease(GERD)is one of refractory digestive disease in clinic characterized by high prevalence,complex and recurrent symptoms.Traditional Chinese medicine(TCM)has a significant effect on the treatment of GERD.Spleen deficiency and damp-heat syndrome becomes one of the main syndrome patterns of GERD adapting with the changes of modern lifestyle.Although the therapy of strengthening the spleen,clearing heat and dampness is widely used in treatment of GERD,further studies still need to be conducted.The article comprehensively analyses the theoretical basis,mechanism and clinical efficacy of the therapy in order to provide more ideas for clinical treatment.

Chinmedomics facilitated quality-marker discovery of Sijunzi decoction to treat spleen qi deficiency syndrome

Sijunzi decoction (SJZD) is a Chinese classical formula to treat spleen qi deficiency syndrome (SQDS) and has been widely used for thousands of years.However,the quality control (QC) standards of SJZD are insufficient.Chinmedomics has been designed to discover and verify bioactive compounds of a variety of formularapidly.In this study,we used Chinmedomics to evaluate the SJZD's efficacy against SQDS to discover the potential quality-markers (q-markers) for QC.A total of 56 compounds in SJZD were characterized in vitro,and 23 compounds were discovered in vivo.A total of 58 biomarkers were related to SQDS,and SJZD can adjust a large proportion of marker metabolites to normal level and then regulate the metabolic profile to the health status.A total of 10 constituents were absorbed as effective ingredients that were associated with overall efficacy.We preliminarily determined malonyl-ginsenoside Rb2 and ginsenoside Ro as the q-markers of ginseng;dehydrotumulosic acid and dihydroxy lanostene-triene-21-acid as the q-markers of poria;glycyrrhizic acid,isoglabrolide,and glycyrrhetnic acid as the q-markers of licorice;and 2-atractylenolide as the q-marker of macrocephala.According to the discovery of the SJZD q-markers,we can establish the quality standard that is related to efficacy.