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.

Recent Situation of Acupuncture and Moxibustion in the Treatment of Stomach Pain with Deficiency and Cold of Spleen and Stomach

Stomachache is one of the most common diseases of digestive system, among which stomach pain of deficiency cold of spleen and stomach is the most common. Acupuncture and moxibustion therapy as an external treatment of traditional Chinese medicine for relieving stomach pain due to deficiency of spleen and stomach, has the advantages of convenient operation, small adverse reactions and remarkable curative effect, which is easy for patients to accept. In this paper, acupuncture and moxibustion therapy such as moxibustion and warm acupuncture therapy for stomach pain of spleen and stomach deficiency cold type is summarized to provide evidence-based basis for clinical treatment of stomach pain of spleen and stomach deficiency cold type.

Discussion on Traditional Chinese Medicine Treatment of Liver-Depression-Spleen-Deficiency Type Insomnia Based on the State-Target Theory and the Regulation of Microbiome-Gut-Brain Axis

Insomnia has become an urgent clinical problem in modern society.Current research has found that the gut flora-gut-brain axis plays an important role in regulating insomnia.The state-target theory is a product of the combination of traditional Chinese medicine and modern molecular biology technology.This paper clarifies the correlation between the state-target theory,the intestinal flora-gut-brain axis,and liver-depression-spleen-deficiency insomnia.The use of traditional Chinese medicine to regulate the structure and abundance of intestinal flora was also explored,aiming to integrate traditional Chinese medicine with Western medicine for the prevention and treatment of liver-depression-spleen-deficiency insomnia.

Clinical Observation of the Therapeutic Effects of Wenzhong Hewei Formula in Treating Spleen and Stomach Qi Deficiency Syndrome in Internal Medicine Diseases

Objective:To observe the clinical efficacy and safety of Wenzhong Hewei Formula in treating spleen and stomach qi deficiency syndrome in internal medicine diseases.Methods:Sixty patients with spleen and stomach qi deficiency syndrome admitted to the hospital from April 2022 to June 2023 were randomly divided into observation and control groups,with 30 patients in each group.The control group received conventional internal medicine treatment,while the observation group was additionally treated with Wenzhong Hewei Formula on the basis of conventional treatment.Both groups were treated for 4 weeks.Results:The total effective rate of treatment in the observation group was higher than that of the control group(P<0.05).After treatment,the traditional Chinese medicine syndrome scores of both groups were significantly lower than before treatment,with the observation group showing a more pronounced reduction(P<0.05).There was no statistically significant difference in the incidence of adverse reactions between the two groups(P>0.05).Conclusion:Wenzhong Hewei Formula can effectively improve clinical symptoms in patients with spleen and stomach qi deficiency syndrome,enhance clinical efficacy,and have a high level of safety,making it worthy of clinical promotion.

Effect of Sijunzi Decoction on the Myonuclear Domain of Rat Soleus in Spleen Qi Deficiency

Objective: To study the mechanism of Sijunzi decoction treating limb weakness in spleen Qi deficiency (SQD) based on the myonuclear domain (MND) theory. Methods: 40 male Sprague-Dawley rats were randomly divided into the normal group, SQD model group (model group), SQD+ still water group (SW group) and SQD+ Sijunzi decoction group (CM group), 10 rats each group;Grip-Strength Meter was used to measure limb grip strength;transmission electron microscope was employed to observe the ultrastructural changes of the myofibers, Image Pro 6.0 was used to measure the myonuclear numbers, cross-section area (CSA) and then their ratios (the MND sizes) were calculated, immunofluorescence assay was chosen to test the expressions of paired box gene 7 (Pax7) and myogenic differentiation antigen (MyoD). Results: Compared with those in the normal group, limb grip strength was decreased, sarcomeres were abnormal, and all the myonuclear numbers, CSA and MND sizes were reduced, but the Pax7+ cell numbers were increased, significantly, in the model and SW groups;Compared with those in the model and SW groups, limb grip strength was increased, sarcomeres were basically normal, the myonuclear number and CSA were both greater, and the Pax7+ and MyoD+ cell numbers were both increased, significantly, in the CM group. Conclusion: Sijunzi decoction might increase the myonuclear number by activating the MSCs to treat limb weakness in SQD.

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.

Clinical Observation on Modified Huan'gan Lipi Decoction Combined with Acupuncture in Treating Spleen Deficiency and Liver Hyperactivity Type Tic Disorders

[Objectives]To observe the clinical efficacy of Modified Huan'gan Lipi Decoction combined with acupuncture in the treatment of spleen deficiency and liver hyperactivity type tic disorders(TD).[Methods]Sixty patients with spleen deficiency and liver hyperactivity type TD were randomly divided into a treatment group of 40 cases and a control group of 20 cases.The treatment group received Modified Huan'gan Lipi Decoction combined with acupuncture,and the control group received Haloperidol Tablets.After 4 weeks of treatment,the Yale Global Tic Severity Scale(YGTSS)score,the total score of TCM syndrome and the clinical efficacy were compared between the two groups before and after treatment.[Results]After treatment,the total effective rate of 87.5%in the treatment group was higher than 80.0%in the control group(P>0.05);the total score of YGTSS and the total score of TCM syndromes in the two groups were compared within groups,P﹤0.01;between groups,P﹤0.01.The recurrence rates of the treatment group and the control group were 11.1%and 71.4%,respectively.The difference between the two groups was statistically significant(P﹤0.01).[Conclusions]Modified Huan'gan Lipi Decoction combined with acupuncture in the treatment of spleen deficiency and liver hyperactivity type TD can significantly improve the patient's tic symptoms,and its long-term efficacy is stable.

Clinical effects of a traditional Chinese medicine nursing programme to intervene in gastric pain of the spleen and stomach with Qi deficiency

BACKGROUND Gastroparesis is a common digestive disorder characterized by delayed gastric emptying,which can lead to symptoms such as nausea,vomiting,abdominal pain,and poor appetite.Traditional Chinese medicine(TCM)has been used for centuries to treat gastrointestinal disorders,including gastroparesis.TCM theory suggests that spleen and stomach qi deficiency syndrome is one of the main pathogenic factors in gastroparesis.Nursing care plays an important role in the treatment of gastroparesis,and TCM nursing interventions have shown promising results in improving patient outcomes.However,there is limited research on the clinical effectiveness of TCM nursing interventions for gastroparesis with spleen stomach deficiency syndrome.This study aimed to evaluate the clinical effect of TCM nursing intervention in the treatment of gastroparesis with spleen stomach deficiency syndrome and to compare it with routine nursing interventions.AIM To analyze the clinical effect of traditional Chinese medicine nursing intervention in the treatment of gastric paraplegia with spleen stomach deficiency syndrome.METHODS From January 2020 to July 2021,80 patients with gastroparesis of spleen stomach qi deficiency type diagnosed in our hospital were selected for the study.The 80 patients were randomly divided into a control group and an experimental group,with 40 cases in each group.During the treatment period,the control group received routine nursing interventions,while the experimental group received traditional Chinese medicine nursing procedures.Compare the nursing effects of the two groups and observe the changes in traditional Chinese medicine symptom scores,pain levels,and sleep quality before and after treatment.RESULTS After treatment,comparing the treatment effects of the two groups,the total effective rate of the experimental group was significantly higher than that of the control group,with statistical significance(P<0.05).There was no statistically significant difference in the TCM symptom score,visual analogue scale(VAS)score,and Pittsburgh sleep quality index(PSQI)score between the two groups before treatment(P>0.05).However,after treatment,the TCM syndrome scores,VAS scores,and PSQI scores of the experimental group were significantly lower than those of the control group,with a statistically significant difference(P<0.05).CONCLUSION In the clinical nursing intervention of patients with mild gastroparesis due to spleen and stomach qi deficiency,the traditional Chinese medicine nursing plan has good clinical application value and nursing effect,and has a good effect on improving patients’pain and sleep quality.

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.

Effect of electro-acupuncture on basic fibroblast growth factor protein and mRNA expression in hippocampal dentate gyrus of spleen deficiency rats

BACKGROUND： Spleen deficiency in traditional Chinese medicine refers to the functional disorder of spleen, pancreas, intestines, and nervous system in modern medicine. OBJECTIVE; To test whether electro-acupuncture could alter basic fibroblast growth factor （bFGF） protein and mRNA expression in the hippocampal dentate gyrus of spleen deficiency rats. DESIGN, TIME AND SETTING： The randomized, controlled, in vivo animal experiment was performed at the National LeveI-B Laboratory of Clinical Cell Molecule and Biology in Shenzhen Hospital of Traditional Chinese Medicine, between March and November in 2008. MATERIALS： Reserpine injection was produced by Guangdong Bangmin Pharmaceutical Co. Rhubarb extract granule preparation was produced by Guangdong Yifang Pharmaceutical. Huanqiu Brand sterile acupuncture pin was provided by Suzhou Acupuncture Supplies, China. Huatuo Brand electroacupuncture instrument （type SDZ-II） was purchased from Suzhou Medical Appliance Factory, China. METHODS： A total of 96 male Sprague Dawley rats were randomly assigned to control （n = 32） and induction （n = 64） groups. Spleen deficiency was induced via intraperitoneal injection of reserpine and intragastric administration of rhubarb. The successful models were randomized into two groups： model and electro-acupuncture, with 32 rats in each group. Electro-acupuncture was administered at Zusanfi （ST 36） and Sanyinjiao （SP 6） acupoints using a condensation wave and rarefaction （condensation wave 15 Hz） at a strength of 6-15 V for 20 minutes, once per day. The appearance of a slight shiver in the corresponding locus was taken as the standard. According to electro- acupuncture time points, each group was assigned to four subgroups at 7, 14, 28, and 49 days, respectively, with eight rats in each subgroup. Immunohistochemical staining, image analysis, and reverse-transcription polymerase chain reaction were performed at different time points. MAIN OUTCOME MEASURES： bFGF protein and mRNA expression in the hippocampal dentate gyrus of spleen deficiency rats. RESULTS： After 7 days of electro-acupuncture therapy, bFGF protein and mRNA expression significantly increased compared with the model and control groups （P 〈 0.05）. After 14 days, bFGF protein and mRNA expression decreased until 28 days, where levels were then equal to the model group and greater than the control group （P 〈 0.05）. After 49 days, the above indices remained increased in the electro-acupuncture group compared to the model and control groups （P 〈 0.05）. CONCLUSION： Continuous electro-acupuncture maintained a high level of bFGF protein and mRNA expression in the hippocampal dentate gyrus of spleen deficiency rats.

Selenium deficiency induces spleen pathological changes in pigs by decreasing selenoprotein expression,evoking oxidative stress,and activating inflammation and apoptosis

Background:The immune system is one aspect of health that is affected by dietary selenium(Se)levels and selenoprotein expression.Spleen is an important immune organ of the body,which is directly involved in cellular immunity.However,there are limited reports on Se levels and spleen health.Therefore,this study established a Se-deficient pig model to investigate the mechanism of Se deficiency-induced splenic pathogenesis.Methods:Twenty-four pure line castrated male Yorkshire pigs(45 days old,12.50±1.32 kg,12 full-sibling pairs)were divided into two equal groups and fed Se-deficient diet(0.007 mg Se/kg)or Se-adequate diet(0.3 mg Se/kg)for 16 weeks.At the end of the trial,blood and spleen were collected to assay for erythroid parameters,the osmotic fragility of erythrocytes,the spleen index,histology,terminal deoxynucleotidyl transferase nick-end labeling(TUNEL)staining,Se concentrations,the selenogenome,redox status,and signaling related inflammation and apoptosis.Results:Dietary Se deficiency decreased the erythroid parameters and increased the number of osmotically fragile erythrocytes(P<0.05).The spleen index did not change,but hematoxylin and eosin and TUNEL staining indicated that the white pulp decreased,the red pulp increased,and splenocyte apoptosis occurred in the Se deficient group.Se deficiency decreased the Se concentration and selenoprotein expression in the spleen(P<0.05),blocked the glutathione and thioredoxin antioxidant systems,and led to redox imbalance.Se deficiency activated the NF-κB and HIF-1αtranscription factors,thus increasing pro-inflammatory cytokines(IL-1β,IL-6,IL-8,IL-17,and TNF-α),decreasing anti-inflammatory cytokines(IL-10,IL-13,and TGF-β)and increasing expression of the downstream genes COX-2 and iNOS(P<0.05),which in turn induced inflammation.In addition,Se-deficiency induced apoptosis through the mitochondrial pathway,upregulated apoptotic genes(Caspase3,Caspase8,and Bak),and downregulated antiapoptotic genes(Bcl-2)(P<0.05)at the mRNA level,thus verifying the results of TUNEL staining.Conclusions:These results indicated that Se deficiency induces spleen injury through the regulation of selenoproteins,oxidative stress,inflammation and apoptosis.

Guipi decoction effects on arginine vasopressin protein and gene expression in the hippocampus, ventromedial hypothalamic nucleus, and prefrontal lobe in rats with spleen deficiency

BACKGROUND： Arginine vasopressin has been shown to enhance learning in experimental animal models. OBJECTIVE： To determine whether Guipi decoction enhances memory and learning by increasing arginine vasopressin levels, and to verify the influence of Guipi decoction on arginine vasopressin protein and gene expression in the hippocampal CA1 region, prefrontal lobe cortex, and ventral nucleus of hypothalamus in rats with spleen deficiency. DESIGN, TIME AND SETTING： The randomized, neuropharmacological, control study was performed in the College of Basic Medical Sciences, Beijing University of Chinese Medicine between March 2002 and March 2005. MATERIALS： Sixty, healthy, male, Wistar rats were used to establish spleen deficiency models according to the traditional Chinese medicine principle of bitter drugs for purgation, improper diet, and overstrain. Arginine vasopressin-1 polyclonal anti-rabbit antibody immunohistochemistry kit and arginine vasopressin in situ hybridization kit were provided by Department of Neuroanatomy in Shanghai Second Military Medical University of Chinese PLA. METHODS： Sixty rats were divided into five groups at random： normal control （n = 11）, model （n = 13）, Guipi decoction （n = 12）, recipe control A （n = 12）, and recipe control B groups （n = 12）. Rats in the latter four groups received 7.5 g/kg of the drugs by intragastric administration each morning, which comprised Dahuang, Houpu, and Zhishi, prepared at a ratio of 2： 1 ： 1. The rats were fasted every other day, but were allowed free access to water at all times. The rats were forced to swim in 25℃ water until fatigued. Rats in the Guipi decoction and two recipe control groups were intragastrically administered 7.5 g/kg Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellets, respectively, each afternoon. Rats in the normal group were intragastrically administered the same amount of normal saline. All rats were treated for 6 weeks. MAIN OUTCOME MEASURES： At 6 weeks after drug administration, rat brain tissues were harvested. Arginine vasopressin-positive protein products and arginine vasopressin mRNA-positive products in the ventral nucleus of hypothalamus, hippocampal CA1 region, and cortex of prefrontal lobe were determined using immunohistochemistry and in situ hybridization, respectively. Image collection and management were performed by means of an Olympus microscope and color image analyzer system. Average absorbencies were measured. RESULTS： Qualitative analysis： in the normal control group, the arginine vasopressin protein- and mRNA-positive cells were round and oval. The immunoreactive products were strongly positive, with a uniform coloration. In the model group, the arginine vasopressin protein- and mRNA-positive ceils exhibited a shortening or disappearance of neuronal processes, as well as cytoplasm that exhibited chromatic non-uniformity. After rats were administrated Guipi decoction, the arginine vasopressin protein- and mRNA-positive cells exhibited rough and long neuronal, and the intensity of cytoplasmic coloration was also enhanced. Quantitative analysis： in the model group, arginine vasopressin protein and mRNA expression was significantly less in the hippocampal CA1 region, cortex of prefrontal lobe, and ventral nucleus of hypothalamus, compared with the normal group. Compared with the model group, the above-mentioned indices were remarkably increased in the Guipi decoction group （P 〈 0.01）, and were more similar to normal levels. Rats treated with Chaihu Shugan powder or Tianwang Buxin pellets displayed increased arginine vasopressin protein and mRNA expression, but the therapeutic effect was inferior to Guipi decoction. CONCLUSION： Qualitative and quantitative analyses demonstrated that the Guipi decoction enhanced arginine vasopressin protein and gene expression in the hippocampal CA1 region, prefrontal lobe cortex, and ventral nucleus of hypothalamus in rats with spleen deficiency.

Effects of Different Processed Products of Radix Codonopsis on Intestinal Flora of Rats with Spleen Deficiency

[Objectives]This study aimed to observe the effects of different processed products of Radix Codonopsis on intestinal flora in rats with spleen deficiency.[Methods]Rat models with spleen deficiency were established by bitter-cold purgation method with Radix et Rhizoma Rhei.Normal group,model group,Radix Codonopsis group,fried Radix Codonopsis group,rice-fried Radix Codonopsis group,honey-fried Radix Codonopsis group and bran-fired Radix Codonopsis group were designed.After subjecting to corresponding treatments,the changes in the quantity of intestinal microorganisms of the rats were detected.[Results]There was no significant change in the quantity of intestinal microorganisms of the rats in the normal group and model group.The rats in the rice-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased,and the quantity of Escherichia coli and Staphylococci reduced.The rats in the Radix Codonopsis group,fried Radix Codonopsis groups,honey-fried Radix Codonopsis group and bran-fried Radix Codonopsis group were administered after successful modeling,and the abundance of Bifidobacterium and Lactobacillus in the intestines of the rats increased(the increases were smaller than those in the rice-fried Radix Codonopsis group),and the abundance of E.coli and Staphylococci reduced,close to normal levels.[Conclusions]Different processed products of Radix Codonopsis have obvious regulation effect on intestinal flora of rats with spleen deficiency,and the regulation effect of rice-fried Radix Codonopsis on rats with spleen deficiency is better than that of Radix Codonopsis,fried Radix Codonopsis,honey-fried Radix Codonopsis and bran-fried Radix Codonopsis.

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.

Guipi decoction effects on brain somatostatin levels and receptor mRNA expression in rats with spleen deficiency

BACKGROUND： Somatostatin is abundant in the hypothalamus, cerebral cortex, limbic system, and mesencephalon. Somatostatin mRNA expression in the brain of rats with spleen deficiency is noticeably reduced, as well as attenuation of cognitive function. OBJECTIVE： To observe the interventional effect of Guipi decoction on somatostatin level and somatostatin receptor 1 （SSTRl） mRNA expression in different encephalic regions of rats with spleen deficiency, and to compare the interventional effects of Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet. DESIGN： A randomized controlled observation. SETTING： Basic Medical College, Beijing University of Traditional Chinese Medicine. MATERIALS： Fifty adult Wistar male rats, of clean grade, weighing （160 ± 10） g, were provided by Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. The protocol was performed in accordance with ethical guidelines for the use and care of animals. Somatostatin 1 polyclonal anti-rabbit antibody and SSTRl in situ hybridization kit were provided by Department of Neuroanatomy, Shanghai Second Military Medical University of Chinese PLA. The drug for developing rat models of spleen deficiency was composed of Dahuang, Houpu and Zhishi, and prepared at 2：1：1. Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet recipes were made according to previous studies. METHODS： This study was performed at the Basic Medical College, Beijing University of Traditional Chinese Medicine from March 2002 to March 2005. The rats were randomly divided into 5 groups, with 10 rats in each group： normal, model, Guipi decoction, Chaihu Shugan powd.er, and Tianwang Buxin pellet groups. Rat models of the latter 4 groups were developed by methods of purgation with bitter and cold nature drugs, improper diet, and overstrain. The rats received 7.5 g/kg of the drugs each morning and were fasted every other day, but were allowed free access to water at all times. The rats were forced to swim in 25 ℃ water until fatigued. Rats in the normal group were intragastrically administered the same amount of normal saline. Rats in the Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet groups were intragastrically administered 7.5 g/kg Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet, respectively, every afternoon. All rats were treated for 6 weeks. MAIN OUTCOME MEASURES： Somatostatin protein and SSTRI mRNA expression in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were determined by immunohistochemistry and in situ hybridization, respectively. RESULTS： Fifty rats were included in the final analysis. In the model group, expression of somatostatin protein and SSTRl mRNA in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were significantly less than in the normal group （P 〈 0.01）. Above-mentioned indices were identical in the Chaihu Shugan powder and model groups. However, expression of somatostatin protein and SSTRl mRNA were significantly higher in the Guipi decoction group compared to model group （P 〈 0.01）. In the Tianwang Buxin pellet group, SSTRl mRNA expression in rat ventral nucleus of hypothalamus and somatostatin level in rat hippocampal CAl region and cortex of prefrontal lobe, as well as ventral nucleus of hypothalamus, were significantly higher compared to model group （P 〈 0.01 ）. CONCLUSION： Somatostatin level and SSTRl mRNA expression in rats with spleen deficiency were lower than in normal rats. Guipi decoction and Tianwang Buxin pellet up-regulated somatostatin level and SSTRl mRNA expression.

Changes of free radicals and digestive enzymes in saliva in cases with deficiency inspleen-yin syndrome

Objective：To explore the nature of deficiency in spleen-yin syndrome,which could provide scientific theoretical support and practical guidance for clinical Traditional Chinese Medicine（TCM）syndrome differentiation based on biology,and had a strong clinical significance.Methods：Serum Cu and Zn were detected by atomic absorption spectrophotometer,serum vitamin E by high performance liquid chromatography,serum vitamin C by 2,4-Dinitrophenylhydrazine Colorimetry,total superoxide dismutase（SOD）and Cu and Zn- SOD by the xanthine oxidase method,and malondialdehyde（MDA）by the 2-thiobarbituric acid method（TBA）.Total antioxidant capacity was detected by a colorimetry kit.Amylase Activity was detected by an automatic biochemical analyzer.Lysozyme was detected by lysozyme detection plate,the diameter of bacteriolysis circle was measured and the corresponding content of lysozyme was obtained from a table of standard curve values.Results： No significant difference in total SOD and Cu,Zn-SOD was found between deficiency in spleen-yin group and normal group.However,such factors in deficiency in kidney-yin group were significantly lower than the other groups（P〈0.05）.The MDA content in both deficiency in spleen-yin group and deficiency in kidney-yin group were significantly higher than that of normal group（P〈0.05）,while the total antioxidant capacity was significantly lower than normal group（P〈0.05）.The vitamin E content in deficiency in kidney-yin group was significantly lower than that in the other two groups（P〈0.05）.No significant difference in the contents of vitamin C,Cu and Zn were observed in these groups.The Zn/Cu level in deficiency in kidney-yin group and the vitamin E level in deficiency in spleen-yin group decreased,but with no significant difference.Amylase activity in unit time in cases with deficiency in spleen-yin was lower than and had significant differences with that in normal cases,and higher than that in cases with deficiency in kidney-yin.The sectional velocity of saliva and the ratio of lysozyme in normal case group were significantly higher than other two groups,while deficiency in the spleen-yin group was significantly higher than the deficiency in kidney-yin group.Conclusion：All the results indicated that the objective pathological mechanism between the deficiency in spleen-yin and deficiency in kidney-yin was different.

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.

Effect of Yiqi Jianpi plus anticancer herbs on spleen deficiency in colorectal cancer and its anti-tumor role

Objective:To observe the effect of Yiqi Jianpi plus anticancer herbs on spleen deficiency in colorectal cancer and its anti-tumor role.Methods:Human intestinal cancer cell HT29 xenograft of nude mice model was established.The expression of ECF,VEGF,gastric cancer tumor growth in mice were observed.Results:Protein kinase C expression in in the Yiqi Jianpi group and Yiqi Jianpi anti-tumor group was significantly better than the model group(P<0.01,P<0.05).There was significantly more apoptotic cells in Yiqi Jianpi anti-tumor group than Yiqi Jianpi group and model group(P<0.01).Epidermal growth factor and vascular endothelial growth factor expression in Yiqi Jianpi group was significantly lower than Yiqi Jianpi group and model group(P<0.05).Conclusions:Tumor can inhibit the expression of PKC inhibition.Yiqi Jianpi and anticancer treatment can reduce this inhibition.Besides this treatment can also inhibit expression of tumor related genes such as epidermal growth factor and vascular endothelial growth factor.

Expression Imbalance of Cholinergic M₂and M₃Receptors Contributes to the Motility Reduction of the Small Intestine in Spleen Qi Deficiency

Objective: To study roles of cholinergic M2 and M3 receptors in the motility reduction of small intestine (SI) in spleen qi deficiency. Methods: 16 male SD rats were randomly divided in the control group and spleen qi deficiency group (model group)—8 rats each group;spleen qi deficiency model of the improper diet and overfatigue was established;the SI propelling rate (SIPR) was used to evaluate the SI motility;ELISA was used to measure concentrations of acetylcholine (ACh), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the SI tissue;immohistochemistry was employed to detect expressions of cholinergic M2 and M3 receptors. Results: Compared with those in the control group, SIPR was reduced;expression of M2 receptors was increased;and expression of M3 receptors and concentrations of cAMP and PKA were decreased, significantly, in the model group. Conclusions: Expression imbalance of cholinergic M2 and M3 receptors might contribute to the motility reduction of the SI in spleen qi deficiency.