Identifications of metabolic differences between Hedysari Radix Praeparata Cum Melle and Astragali Radix Praeparata Cum Melle for spleen-qi deficiency rats: A comparative study.

Hedysari Radix Praeparata Cum Melle (HRPCM) and Astragali Radix Praeparata Cum Melle (ARPCM) are capable of improving spleen-qi deficiency (SQD) syndrome especially in the gastrointestinal dysfunction and decreased immunity in traditional Chinese medicine clinically. This study aims to compare and reveal the metabolic differences between HRPCM and ARPCM for SQD rats. Firstly, HRPCM (12.6 g/kg) and ARPCM (12.6 g/kg) were used to intervene SQD rats to further evaluate the effect. The results showed that HRPCM and ARPCM were able to improve the spleen pathology, increase the body weight, the rectal temperature, the spleen index, the thymus index, the levels of GAS and D-xylose in serum, and decrease the levels of IL-2, IL-6 and TNF-α in serum for SQD rats. Then, the studies of metabolic differences in serum and spleen were carried out using UPLC-Q-TOF-MS. The findings emphasized that HRPCM and ARPCM not only regulated metabolic profiling of serum and spleen in SQD rats, but also existed differences. HRPCM and ARPCM regulated metabolic pathways mainly including lipid metabolism, energy metabolism, amino acid metabolism, nucleotide metabolism, sugar metabolism and other types of metabolism for SQD rats. However, the metabolite profiles in SQD rats changed significantly, mainly involving abnormal glycine synthesis occurred in SQD rats. The expression trends of metabolites in HRPCM and ARPCM intervention for SQD rats were partly the same. Interestingly, there are similarities and differences in metabolic profiling between HRPCM and ARPCM for SQD rats. The differences were mainly in the synthesis of L-glutamine in amino acid metabolism.

Urine and serum metabolomics study of wild ginseng in rats with spleen-qi deficiency using rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry.

Patients with a spleen-qi deficiency often exhibit dysfunction in the metabolic system. Metabolites are considered the most direct reflection of individual physiological and pathological conditions and represent attractive candidates to provide deep insights into disease phenotypes. This study examines the potential therapeutic mechanism of wild ginseng on spleen-qi deficiency through the analysis of serum and urine metabolomics using rapid-resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. The reasons for the superiority of wild ginseng treatment over cultivated ginseng were also analyzed in depth. After wild ginseng intervention, anandamide, urobilinogen, aldosterone, and testosterone glucuronide were significantly reduced in serum. Meanwhile, argininosuccinic acid, L-cysteine, and seven other metabolites were significantly elevated in serum. Nine metabolites, including L-acetylcarnitine, and citrulline were elevated in the urine, and trimethylamine N-oxide, adrenic acid, and 10 other metabolites were reduced. Arginine biosynthesis, pantothenate and CoA biosynthesis, thiamin metabolism, taurine, and tryptophan metabolism pathways were mainly improved. Further analysis was conducted on the relationship between Lactobacillus and Akkermansia bacteria with metabolites, and it was found that they are mainly related to amino acid metabolites. This study provides strong theoretical support and direction for further explanation of the immune mechanism of wild ginseng and lays the foundation for future studies.

A screening strategy for bioactive components of Bu-Zhong-Yi-Qi-Tang regulating spleen-qi deficiency based on "endobiotics-targets-xenobiotics" association network.

ETHNOPHARMACOLOGICAL RELEVANCE: Bu-Zhong-Yi-Qi-Tang is a famous traditional Chinese medicine formula that has been prevalent in China for over 700 years to treat spleen-qi deficiency related diseases, such as gastrointestinal and respiratory disorders. However, the bioactive components responsible for regulating spleen-qi deficiency remain unclear and have puzzled many researchers. AIM OF THE STUDY: The current study focuses on efficacy evaluation of regulating spleen-qi deficiency and screening the bioactive components of Bu-Zhong-Yi-Qi-Tang. MATERIALS AND METHODS: The effects of Bu-Zhong-Yi-Qi-Tang were evaluated through blood routine examination, immune organ index, and biochemical analysis. Metabolomics was utilized to analyze the potential endogenous biomarkers (endobiotics) in the plasma, and the prototypes (xenobiotics) of Bu-Zhong-Yi-Qi-Tang in the bio-samples were characterized using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Then, these endobiotics were used as "bait" to predict targets based on network pharmacology and to screen potential bioactive components from the absorbed prototypes in the plasma by constructing an "endobiotics-targets-xenobiotics" association network. Further, the anti-inflammatory activities of representative compounds (calycosin and nobiletin) were validated through poly(I:C)-induced pulmonary inflammation mice model. RESULTS: Bu-Zhong-Yi-Qi-Tang exhibited immunomodulatory and anti-inflammatory activities in spleen-qi deficiency rat, as supported by the observation of increased levels of D-xylose and gastrin in serum, an increase in the thymus index and number of lymphocytes in blood, as well as a reduction in the level of IL-6 in bronchoalveolar lavage fluid. Furthermore, plasma metabolomic analysis revealed a total of 36 Bu-Zhong-Yi-Qi-Tang related endobiotics, which were mainly enriched in primary bile acids biosynthesis, the metabolism of linoleic acid, and the metabolism of phenylalanine pathways. Meanwhile, 95 xenobiotics were characterized in plasma, urine, small intestinal contents, and tissues of spleen-qi deficiency rat after Bu-Zhong-Yi-Qi-Tang treatment. Using an integrated association network, six potential bioactive components of Bu-Zhong-Yi-Qi-Tang were screened. Among them, calycosin was found to significantly reduce the levels of IL-6 and TNF-α in the bronchoalveolar lavage fluid, increase the number of lymphocytes, while nobiletin dramatically decreased the levels of CXCL10, TNF-α, GM-CSF, and IL-6. CONCLUSION: Our study proposed an available strategy for screening bioactive components of BYZQT regulating spleen-qi deficiency based on "endobiotics-targets-xenobiotics" association network.

Effect of physical rehabilitation combined with Xiaoqinglong Decoction on the syndrome of lung and spleen deficiency in children with asthma remission / 西安交通大学学报(医学版)

【Objective】 To investigate the effect of physical therapy combined with traditional Chinese medicine (TCM) in treating the syndrome of lung and spleen qi deficiency in children at asthma remission stage and the effect on serological indicators. 【Methods】 A total of 130 children with asthma in remission treated in our hospital from May 2018 to May 2020 were selected and randomly divided into combination group (n=65) and physical group (n=65). The physical group was treated with physical therapy and Montelukaste sodium chewable tablet, while the combination group was treated with Xiaoqinglong Decoction based on soil preparation and gold generation method. Both groups were treated for three months. The scores of chest tightness (attack), cough, asthma and laryngeal wheezing were compared between the two groups before and after treatment. The average score of TCM syndrome was recorded before treatment (T0), 1, 2 and 3 months after treatment (T1, T2 and T3), and 6 and 9 months after treatment (T4 and T5). Serum levels of interleukin 6 (IL-6), immunoglobulin (IgE) and tumor necrosis factor α(TNF-α) were detected before and after treatment. 【Results】 After treatment, the total effective rate in the combination group (86.15%) was higher than that in the physical group (73.85%) (χ2=4.333, P<0.05). After treatment, the TCM syndrome scores of asthma, laryngeal phlegm were significantly lower in the combination group than in the physical group (P<0.05). The average score of TCM syndrome in T3, T4 and T5 combination group was lower than that in the physical group (t=3.167, 3.317, 4.168, 4.267, P<0.05). After treatment, the levels of IL-6, IgE and TNF-α were decreased (P<0.05), and the levels of IgE and TNF-α in the combination group were lower than those in the physical group (t=12.164, 5.136, P<0.05). 【Conclusion】 Physical therapy combined with Xiaoqinglong Decoction can improve clinical symptoms and regulate immunity of children at asthma remission stage.

Distribution of Traditional Chinese Medicine Patterns and Analysis of Factors Related to Acute Exacerbation in Group E of Chronic Obstructive Pulmonary Disease / 中医杂志

ObjectiveTo analyze the distribution of traditional Chinese medicine （TCM） patterns as well as factors related to acute exacerbation in group E of chronic obstructive pulmonary disease （COPD）. MethodsThe general data of 161 COPD patients， including gender， age， body mass index （BMI）， disease course， smoking history， and past history， were collected. In terms of the four examinations of TCM， the differentiated patterns included phlegm-heat obstructing the lung， turbid phlegm obstructing the lung， phlegm stasis obstructing the lung， lung-spleen qi deficiency， and lung-kidney deficiency. The modified British Medical Research Council （mMRC） scale and COPD assessment test （CAT）， the pulmonary function indicators including forced expiratory volume in the first second （FEV1） and ratio of forced expiratory volume to forced vital capacity at second 1 （FEV1/FVC）， GOLD grade， and the patient's acute exacerbations in the previous year were recorded. Multivariate regression analysis was performed using logistic regression model to determine the relevant factors of patients in COPD group E. The distribution of acute exacerbations in different TCM symptom patients in group E was analyzed. ResultsThere were 80 patients （49.69%） in group E and 81 patients （50.31%） in non-group E. In group E， 23 （28.75%） patients had a history of two acute exacerbations， while 35 （43.75%） had three acute exacerbations， and 22 （27.5%） had more than three acute exacerbations. There were 13 （16.25%） cases of phlegm-heat obstructing the lung pattern， 6 （7.5%） cases of turbid phlegm obstructing the lung pattern， 8 （10%） cases of phlegm stasis obstructing the lung pattern， 22 cases （27.5%） of lung-spleen qi deficiency pattern， and 31 （38.75%） cases of lung-kidney deficiency pattern. There were significant differences in smoking history， disease course， TCM pattern， TCM syndrome score， mMRC score， and CAT score between groups （P＜0.05）. A total of 107 of the 161 patients completed pulmonary function tests， and the differences in FEV1， FEV1/FVC and GOLD grades between groups were statistically significant （P＜0.05）. Multivariate regression analysis showed that TCM pattern， TCM syndrome score and CAT score were statistically significant factors for COPD patients in group E （P＜0.05）. There were statistically significant differences in the number of acute exacerbations in different TCM patterns in group E （P＜0.05）. The patients with two acute exacerbations in the past year were mainly phlegm-heat obstructing the lung and lung-spleen qi deficiency patterns， while the three acute exacerbations were mainly seen in lung-spleen qi deficiency and lung-kidney deficiency patterns， and more than three exacerbations were more common with lung -kidney deficiency pattern. ConclusionsPatients in COPD group E were mainly the lung-spleen qi deficiency and lung-kidney deficiency patterns. Deficiency of healthy qi is the main reason for the increase in the number of acute exacerbations， and TCM patterns and CAT score were the main related factors.

Influence of spleen-qi deficiency syndrome in type 2 diabetes mellitus on Ankle-Brachial Index / 国际中医中药杂志

Objective:To investigate the influence of T2DM Spleen-qi Deficiency syndrome on Ankle-Brachial Index (ABI).Methods:The clinical data of 298 patients with T2DM who were hospitalized in Dongzhimen Hospital, Beijing University of Chinese Medicine from January 2019 to December 2021 were retrospectively analyzed. According to the diagnostic criteria of spleen-qi deficiency syndrome, the patients were divided into two groups: spleen-qi deficiency syndrome group and non-spleen-qi deficiency syndrome group. There were 142 patients in the spleen-qi deficiency syndrome group and 156 patients in the non-spleen-qi deficiency syndrome group. The differences of ABI between the two groups were compared, and the correlation between spleen-qi deficiency syndrome and clinical indicators (gender, age, body mass index, course of diabetes, history of hypertension, smoking history, fasting glucose, total cholesterol, triglyceride, platelet, hemoglobin, serum albumin, blood urea nitrogen, serum creatinine, estimated glomerular filtration rate) and ABI in T2DM patients was analyzed.Results:The left ABI [1.09 (1.00, 1.19) vs. 1.13 (1.03, 1.22)] and the right ABI [1.09 (0.96, 1.17) vs. 1.12 (1.02, 1.20)] in T2DM spleen-qi deficiency syndrome group were significantly lower than those in non-spleen-qi deficiency group ( P<0.05).The left ABI was significantly correlated with spleen-qi deficiency syndrome ( r=0.122, P=0.035) and estimated glomerular filtration rate ( r=0.137, P=0.018), and the right ABI was significantly correlated with spleen-qi deficiency syndrome ( r=0.123, P=0.034) and PLT ( r=-0.115, P=0.047). After correcting for other confounding factors by multiple linear regression analysis, there was significantly correlation between spleen-qi deficiency syndrome and ABI. Conclusion:Compared with the non-spleen-qi deficiency syndrome group, T2DM patients in the spleen-qi deficiency group had a lower ankle-brachial index and were more likely to develop peripheral arterial disease.

Establishment and Evaluation of Rat Model for Chronic Obstructive Pulmonary Disease with Lung-spleen Qi Deficiency / 中国实验方剂学杂志

ObjectiveTo establish and evaluate a chronic obstructive pulmonary disease （COPD） model with lung-spleen qi deficiency. MethodA rat model mimicking COPD with lung-spleen qi deficiency was established by the combination of cigarette smoking and intratracheal instillation of lipopolysaccharide （LPS） along with gavage of Sennae Folium infusion. Forty male SPF-grade SD rats were randomly assigned to blank， model， and low- （L-FXY）， medium- （M-FXY）， and high-dose （H-FXY） Sennae Folium infusion groups. Other groups except the blank group were exposed to daily cigarette smoke， with LPS administrated via intratracheal instillation on the 1st and 14th days. On the 28th day of modeling， the L-FXY， M-FXY， and H-FXY groups were administrated with Sennae Folium infusion at 5， 10， and 20 g·kg-1， respectively， and at 4 ℃ for three weeks. The modeling lasted for 49 days. The general conditions （body mass， food intake， fecal water content， and anal temperature） and behaviors （grip strength test and tail suspension test） of rats in different groups were examined. The lung function， lung histopathology， D-xylose， amylase， and gastrin levels in the serum， interleukin（IL）-1β and IL-6 levels in the alveolar lavage fluid， levels of T-lymphocyte subsets （CD4+， CD8+， and CD4+/CD8+） in the peripheral blood， and thymus and spleen indices were measured. ResultTwo rats died in the H-FXY group. Compared with the blank group， both the M-FXY and H-FXY groups exhibited reduced body mass and food intake （P<0.01） and increased fecal water content （P<0.01）. The anal temperature in the H-FXY group was lower than that in the blank group （P<0.01）. The grip strength decreased in the modeling groups compared with the blank group （P<0.01）， and the duration of immobility in the tail suspension test increased in the M-FXY and H-FXY groups （P<0.05， P<0.01）. Compared with the blank group， the modeling groups showed reduced 0.3 second forced expiratory volume （FEV0.3）， FEV0.3/forced vital capacity （FVC）（P<0.01）， thickening of bronchial walls， proliferation of goblet cells， and the presence of emphysematous changes. In terms of gastrointestinal function， the M-FXY and H-FXY groups had lower levels of D-xylose， gastrin， and α-amylase than the blank group （P<0.01）. Regarding the immune and inflammatory indices， the M-FXY and H-FXY groups showed lower thymus and spleen indices than the blank group （P<0.01）. Compared with the blank group， the modeling groups presented lowered CD4+ level （P<0.01） and CD4+/CD8+ ratio （P<0.05， P<0.01） in the peripheral blood and elevated levels of IL-1β and IL-6 in the alveolar lavage fluid （P<0.01） than the blank group. ConclusionA model of COPD with lung-spleen Qi deficiency was established through the combination of daily cigarette smoke， intratracheal instillation with LPS， and gavage of Sennae Folium infusion. The comprehensive evaluation results suggested medium-dose （10 g·kg-1） Sennae Folium infusion for gavage during the modeling of COPD with lung-spleen Qi deficiency.

Metabolomics and network pharmacological analysis to explore the mechanism of ginseng treatment for spleen-qi deficiency.

Ginseng is the main Chinese herbal medicine for tonifying Qi and invigorating the spleen. It has been used to treat spleen-qi deficiency with good protective effects for thousands of years, however, its biological mechanism has not been fully elucidated. This study aims to explore the mechanism of ginseng in the treatment of spleen-qi deficiency by using a comprehensive method combining metabolomics and network pharmacological analysis. Gas chromatography-mass spectroscopy was applied for investigating the changes in urine metabolites in spleen-qi deficiency rats and after treatment with ginseng. Metabolomics and network pharmacology analysis were applied to screen potential biomarkers and therapeutic targets of ginseng in the treatment of spleen-qi deficiency, respectively. Molecular docking was employed to further evaluate the docking mode of potential biomarkers and therapeutic target proteins. The results of metabolomics showed that the therapeutic effects of ginseng are mainly related to its regulation of three metabolic pathways. The molecular structure of potential biomarkers and common proteins was further analyzed by molecular docking to verify its effectiveness. Ginseng has good pharmacological effects by controlling key targets of related metabolic pathways, signal pathways, and potential biomarkers.

Intervention effects of ginseng on spleen-qi deficiency in rats revealed by GC-MS-based metabonomic approach.

Panax ginseng C. A. Mey. (Ginseng) is a famous Chinese medicine with tonifying middle and replenishing qi effects and has been applied for the treatment of spleen-qi deficiency for many years. However, its potential therapeutic mechanisms have not been thoroughly studied. In this study, the metabolomic technique was applied to explore the therapeutic effect of ginseng on the spleen-qi deficiency. A rat model of spleen-qi deficiency was generated via the fatigue swimming method. After 3 weeks of treatment with ginseng, the entire metabolic changes in rat serum were profiled by gas chromatography-mass spectroscopy (GC-MS). The metabolic profiles in serum taurine and hypotaurine metabolism significantly differed among groups, in which a total of 17 metabolites were identified. Ginseng reversed the metabolic changes in the difference involving some metabolic pathways. Among them, beta-alanine metabolism, taurine and hypotaurine metabolism, and the pentose phosphate pathway are the key metabolic pathways. The therapeutic effects of ginseng on spleen-qi deficiency rats could be achieved by regulating multiple metabolic pathways, metabolites can be used as potential biomarkers for the diagnosis and monitoring of spleen-qi deficiency.

The effects of the recommended prescription for COVID-2019-lung-spleen qi deficiency on in vitro SARS-CoV-2 replication and pro-inflammatory factors expression / 中国药理学通报

Aim To determine the antiviral and anti-inflammatory effects of the recommended prescription for COVID-2019-lung-spleen qi deficiency(4-1)against in vitro infection of SARS-CoV-2 and common coronaviruses.Methods The main chemical substances of 4-1 were analyzed by LC-MS.The toxicity and antiviral effects of of 4-1 were detected by MTT and by CPE assay, respectively.The viral loads in cell supernatant and the expression of inflammatory factors induced by viral infection were determined by qRT-PCR.Results The recommended prescription 4-1 contained 94 chemical compounds, including flavonoids, steroids, sesquiterpenoids, and so on.The range of selection indexes for SARS-CoV-2 and common coronaviruses was 8.44±0.4952.26±2.3.This prescription could inhibit the proliferation of SARS-CoV-2, the expression of ACE2 and S mRNA, and down regulate IL-1α and CCL-5/RANTES at 10, 5, and 2.5 g•L-1 doses.Further, at doses of 20, 10 and 5 g•L-1, it could inhibit the proliferation of three common coronaviruses and suppress the overexpression of IL-6, CXCL-8/IL-8, CXCL-10/IP-10, TNF-α, IFN-α, CCL-2/MCP-1, MIG and CCL-5/RANTES induced by OC43/229E infection.The inhibitory effects were dose-dependent.Conclusions The prescription 4-1 has antiviral and anti-inflammation effects against multiple coronaviruses.This study provides the research basis for the treatment of common respiratory viral infections and emerging infectious diseases such as COVID-19 by using traditional Chinese medicine.

[Comparative study of Astragali Radix Praeparata cum Melle and Hedysari Radix Praeparata cum Melle on spleen Qi deficiency rats].

This Radix study cum aims Melle to explore(HRPM)the on efficacy spleen differences deficiency between syndrome.modeling Astragali A Radix of Praeparata110cum rats Melle were(ARPM)randomized fatigue),and into rats Hedysari a Praeparata(n qi total irregular HRPM male diet,SD diarrhea,control were(n Yiqi=10)the=100).Pill group fied and model a modeling group,group Buzhong After(BYP)(through ARPM and the HRPM-H),classimedium-dose into(ARPM-M raised group,and high-dose(ARPM-H each and Rats BYP and under HRPM-M),normal and low-dose and(ARPM-L in and group HRPM-L)were groups,continuously10rats induced.were in group.the in group the were18.9,control given group were g·kg~(-1)conditions while those the the model Rats respectively in18.912.6,BYP kg~(-1)water extract,decoction those in ARPM/HRPM-H,the-M,dosage lasted and of-L groups treated the with control and model6.3group g·rewere motilin determined m L·kg~(-1)·day~(-1).days.of dose Spleen ARPM/HRPM of in water.morning,The at the10Rats spleen in index group thymus and index ceived equal calculated.(MTL),distilled tissue administration to15observe Then the and Routine of each group D-xylose,were was(IL-2),the subjected HE stainingγ(IFN-γ),lower to the pathological changes.(IgA),blood gastric indexes,mucosa index,interleukin-2group.interferon group immunoglobulin of A and spleen pepsin index,of in Ig A,IL-2spleen IFN-γ,control each MTL,levels Rats pepsin the in model(P<0.01),had higher levels routine(P<0.01),blood and indexes,more thymus lesions D-xylose,the and in index,level decreased HRPM-L of IL-2severe compared spleen with than the those model in group.thymus group.that(P<0.05group,P<0.01)index administration thymus groups Ig A or spleen as that and in spleen routine Except index,spleen the Ig A,index,group and were in in ARPM-M model group,group,index,indexes,P<0.01)and thymus MTL index,those in ARPM-L insignificantly Ig A,different pepsin from other those in the the blood index,compared IFN-γ,group,(P<0.05The D-xylose,model MTL,spleen and lesions high-dose in each administration administration groups group increased relieved.blood or comparison as of with HRPM in as the folARPM and the effect in and were white and result than ARPM and is of lows:(P stronger<0.05),of medium-dose high-dose HRPM HRPM on IL-2cell high-dose of(WBC)and count medium-dose the HRPM and corresponding doses than IFN-γmore ARPM the obvious effect(P<0.05of on evident(P<0.05of impact P<0.01),on low-dose between the on corresponD-xylose P<0.01),doses ding MTL doses than Meanwhile,in of or more high-dose,and medium-dose,difference HRPM the and indexes.corresponding there of ARPM in or IL-2no levels in the HRPM-L effect and two groups,on but conclusion,other the both functions IFN-γwas group no was difference more the than recovery that of the and ARPM-H between(IL-2,P<0.01;ARPM-L recovery HRPM the IFN-γ,P<0.05).HRPM-H and obvious therapeutic in rats group qi In ARPM dose have are certain equivalent,effects on with spleen function deficiency.the Specifically,is the better difference immunomodulatory of two at g·low kg~(-1).and but the promote immunomodulatory the of former rats significantly ARPM.than that between of the later two at in the dose>18.9HRPM promotion can of better digestion digestion absorption and may absorption due of than The immunoregulation and be to the difference in clinical medication.

Efficacy of Sijunzi decoction on limb weakness in spleen Qi deficiency model rats through adenosine monophosphate-activated protein kinase/unc-51 like autophagy activating kinase 1 signaling.

OBJECTIVE: To investigate the efficacy of Sijunzi decoction () on limb weakness in a rat model of spleen Qi deficiency (SQD), and to study its effect on mitophagy in skeletal muscle through adenosine monophosphate-activated protein kinase (AMPK) / unc-51 like autophagy activating kinase 1 (ULK1) signaling. METHODS: SQD model rats were produced by fasting combined with forced swimming method for 15 d. After model assessment, rats were randomly divided into four groups of 10 [low/middle/high (L/M/H) Sijunzi decoction dose groups and a normal saline (S) group]. Limb holding power (HP) and body mass (BM) were measured after 2 weeks of treatment. Following euthanasia, quadriceps femoris were dissected and myofiber and mitochondrial morphology were observed by transmission electron microscopy (TEM). Mitochondrial membrane potential (MMP), adenosine triphosphatase (ATP) and reactive oxygen species (ROS) levels were determined using colorimetric methods, and immunoblot analysis of Microtubule-associated protein light chain 3 (LC3) and Sequestosome 1 (p62) was performed to monitor mitophagy and AMPK/ULK1 signaling. RESULTS: Compared with control (C) group rats, in the S group, HP was reduced, the myofiber Z line was disordered, mitochondria were scattered, and numerous vacuoles and mitophagy were observed. MMP and ATP levels were reduced, ROS levels were elevated, and LC3B expression, and p-AMPKα (Thr172)/AMPKα, p-ULK1 (Ser555)/ULK1, and p-Raptor (Ser792)/Raptor ratios were increased, while p62 expression and p-mTOR (Ser2448)/mTOR and p-ULK1 (Ser757)/ULK1 ratios were decreased. After treatment, compared with the S group, HP was improved in M and H groups but not in the L group. Mitophagy was reduced in M, H and L groups but the Z line was disordered and vacuolization remained in the L group. ATP levels were elevated in M, H and L groups, and MMPs were elevat-ed in M and H groups but not in the L group. ROS levels were decreased in M, H and L groups, as were LC3B expression and p-Raptor (Ser792)/Raptor ratios, while p62 expression and p-mTOR (Ser2448)/mTOR and p-ULK1 (Ser757)/ULK1 ratios were increased in M and H groups but not in the L group. p-AMPKα (Thr172)/AMPKα and p-ULK1 (Ser555)/ULK1 ratios were decreased in M, H and L groups. CONCLUSIONS: Sijunzi decoction improved HP, possibly by inhibiting mitophagy via suppression of AMPK/ULK1 signaling. This restored mitochondrial morphology and improved oxidative phosphorylation, which contributed to recovery of limb weakness in SQD model rats.

Comparative study of Astragali Radix Praeparata cum Melle and Hedysari Radix Praeparata cum Melle on spleen Qi deficiency rats / 中国中药杂志

This Radix study cum aims Melle to explore(HRPM)the on efficacy spleen differences deficiency between syndrome.modeling Astragali A Radix of Praeparata110cum rats Melle were(ARPM)randomized fatigue),and into rats Hedysari a Praeparata(n qi total irregular HRPM male diet,SD diarrhea,control were(n Yiqi=10)the=100).Pill group fied and model a modeling group,group Buzhong After(BYP)(through ARPM and the HRPM-H),classimedium-dose into(ARPM-M raised group,and high-dose(ARPM-H each and Rats BYP and under HRPM-M),normal and low-dose and(ARPM-L in and group HRPM-L)were groups,continuously10rats induced.were in group.the in group the were18.9,control given group were g·kg~(-1)conditions while those the the model Rats respectively in18.912.6,BYP kg~(-1)water extract,decoction those in ARPM/HRPM-H,the-M,dosage lasted and of-L groups treated the with control and model6.3group g·rewere motilin determined m L·kg~(-1)·day~(-1).days.of dose Spleen ARPM/HRPM of in water.morning,The at the10Rats spleen in index group thymus and index ceived equal calculated.(MTL),distilled tissue administration to15observe Then the and Routine of each group D-xylose,were was(IL-2),the subjected HE stainingγ(IFN-γ),lower to the pathological changes.(IgA),blood gastric indexes,mucosa index,interleukin-2group.interferon group immunoglobulin of A and spleen pepsin index,of in Ig A,IL-2spleen IFN-γ,control each MTL,levels Rats pepsin the in model(P<0.01),had higher levels routine(P<0.01),blood and indexes,more thymus lesions D-xylose,the and in index,level decreased HRPM-L of IL-2severe compared spleen with than the those model in group.thymus group.that(P<0.05group,P<0.01)index administration thymus groups Ig A or spleen as that and in spleen routine Except index,spleen the Ig A,index,group and were in in ARPM-M model group,group,index,indexes,P<0.01)and thymus MTL index,those in ARPM-L insignificantly Ig A,different pepsin from other those in the the blood index,compared IFN-γ,group,(P<0.05The D-xylose,model MTL,spleen and lesions high-dose in each administration administration groups group increased relieved.blood or comparison as of with HRPM in as the folARPM and the effect in and were white and result than ARPM and is of lows:(P stronger<0.05),of medium-dose high-dose HRPM HRPM on IL-2cell high-dose of(WBC)and count medium-dose the HRPM and corresponding doses than IFN-γmore ARPM the obvious effect(P<0.05of on evident(P<0.05of impact P<0.01),on low-dose between the on corresponD-xylose P<0.01),doses ding MTL doses than Meanwhile,in of or more high-dose,and medium-dose,difference HRPM the and indexes.corresponding there of ARPM in or IL-2no levels in the HRPM-L effect and two groups,on but conclusion,other the both functions IFN-γwas group no was difference more the than recovery that of the and ARPM-H between(IL-2,P<0.01;ARPM-L recovery HRPM the IFN-γ,P<0.05).HRPM-H and obvious therapeutic in rats group qi In ARPM dose have are certain equivalent,effects on with spleen function deficiency.the Specifically,is the better difference immunomodulatory of two at g·low kg~(-1).and but the promote immunomodulatory the of former rats significantly ARPM.than that between of the later two at in the dose>18.9HRPM promotion can of better digestion digestion absorption and may absorption due of than The immunoregulation and be to the difference in clinical medication.

Application of RRLC-QTOF-MS-based metabonomics and UPE for investigating Spleen-Qi deficiency syndrome with Panax ginseng treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Spleen-Qi deficiency is a syndrome of traditional Chinese medicine. Panax ginseng (ginseng) is well known as the key herb for replenishing Qi and tonifying Spleen. However, the pathogenesis of Spleen-Qi deficiency syndrome and therapeutic mechanism of ginseng on Spleen-Qi deficiency constitution have not yet been entirely elucidated. AIM OF THE STUDY: The aim of this work was to investigate the pathogenesis of Spleen-Qi deficiency syndrome and therapeutic mechanism of ginseng on Spleen-Qi deficiency constitution. MATERIALS AND METHODS: The urinary metabonomics was investigated before and after ginseng treatment in Spleen-Qi deficiency subjects by rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-QTOF-MS). The spectra of metabolites were analyzed by principal component analysis (PCA) and the score showed significant difference among the three different groups. The serum biochemical parameters creatine kinases (CK), lactate dehydrogenase (LDH), interferon-γ (INF-γ), Interleukin-4 (IL-4), superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by assay kits. In addition, ultraweak photon emission (UPE) intensity was measured at dazhui point in each subject. RESULTS: According to the orthogonal partial least-squares discriminant analysis (OPLS-DA) results and biochemistry databases searching, 15 potential biomarkers were identified to be involved in Spleen-Qi deficiency syndrome and ginseng influenced Spleen-Qi deficiency. The metabonomics and biological experiment data indicated that metabolism pathways were corresponding to energy metabolism, amino acid, carbohydrate, lipid and pyrimidine metabolisms, oxidative stress urea cycle, and intestinal flora metabolism. And the UPE intensity decreased significantly after the treatment. CONCLUSIONS: Ginseng could promote the related metabolisms in Spleen-Qi deficiency constitution and the metabolites and UPE intensity may be useful as potential biomarkers for diagnosing and monitoring for Spleen-Qi deficiency syndrome.

[Untargeted metabonomics study of ginseng in treatment of spleen-Qi deficiency].

Ginseng has been used to treat Qi-deficiency syndrome up to now, while the therapeutic mechanism is still unclear. In order to explore the mechanism of ginseng in the treatment of Qi-deficiency constitution, the untargeted metabonomics with blood was studied based on rapid resolution high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(RRLC-Q-TOF-MS). In the results, 13 potential biomarkers were found and identified, which mainly involved in the body's antioxidant and immune functions and energy, glycerol, fatty acid, sugar metabolism and bile acid metabolism. The results of blood biochemical analysis also indicated that ginseng could regulate the body's energy metabolism, immune functions and antioxidant capacity in spleen-Qi deficiency constitution. This study revealed the mechanism of ginseng in the treatment of spleen-Qi deficiency using the blood metabonomics, which could provide technological support and scientific basis for further research on ginseng treatment of Qi-deficiency.

A metabonomics and lipidomics based network pharmacology study of qi-tonifying effects of honey-processed Astragalus on spleen qi deficiency rats.

Honey-processed Astragalus is a dosage form of radix Astragali processed with honey, which is deemed to contain better qi-tonifying effects in traditional Chinese medicine theroy. Our previous study has demonstrated that honey-processed Astragalus exhibited a better effect on reinforcing qi (vital energy) and immune improvement toward spleen qi deficiency compared with radix Astragali. However, the detailed mechanisms related to qi-tonifying effects of honey-processed Astragalus is still unclear. In this study, we evaluated the qi-tonifying effects of honey-processed Astragalus on spleen qi deficiency rats and predicted the mechanisms by aggregating metabonomics, lipidomics and network pharmacology. The results revealed that body weights, symptom scores, the levels of red blood cell, white blood cell, lymphocyte, spleen and thymus indexes, and three cytokines (TNF-α, IL-6, IFN-γ) in honey-processed Astragalus treated rats were improved in comparison with spleen qi deficiency rats. In parallel, based on the 26 biomarkers screened in metabonomics and lipidomics, we inferred that glycerophospholipid metabolism significantly regulated in pathway analysis was connected with qi-tonifying effects. Moreover, the network pharmacology analysis concluded that the compounds targets of honey-processed Astragalus CDK2, NOS3, MAPK14, PTGS1 and PTGS2 interacted with markers targets PLA2G(s) family and LYPLA1 could be responsible for regulation of glycerophospholipid metabolism to develop qi-tonifying effects. What's more, the above processes were possibly through VEGF signaling and MAPK signaling pathways.

[Effects of electroacupuncture at "Zusanli"(ST36) on ultrastructure and mitochondrial dynamics of skeletal muscle in rats with spleen qi deficiency syndrome].

OBJECTIVE: To observe the effects of electroacupuncture (EA) at "Zusanli" (ST36) on the ultrastructure and mitochondrial dynamics of skeletal muscle tissue in spleen qi deficiency rats, so as to explore the partial action mechanism of EA at ST36 for spleen deficiency syndrome. METHODS: Twenty-four male SD rats were randomly divided into 4 groups: normal group, model group, ST36 group and non-acupoint group (n＝6 in each group). The model of spleen qi deficiency syndrome was established by improper diet and exhaustive swimming. EA (2 Hz/15 Hz, 0.5 mA) was applied to bilateral ST36 in the ST36 group and non-acupoint in the non-acupoint group for 20 min, once daily for 7 days. The colorimetric method was used to detect the ATP content in skeletal muscle tissue. The ultrastructure changes of skeletal muscle tissue were observed by transmission electron microscopy. The expression levels of optic atrophy 1 (Opa1) and dynamin-related protein 1 (Drp1) mRNA and proteins in the skeletal muscle tissue were determined by fluorescence quantitative real-time PCR and Western blot, respectively. RESULTS: The ATP content in skeletal muscle tissue of model group was significantly lower than that in the normal group (P<0.05), while significantly higher in the ST36 group than that in the model group and non-acupoint group (P<0.05). Transmission electron microscopy showed that a large number of muscle fibers that were ruptured, damaged, and disorganized; moreover, many vacuoles with different sizes, and abnormally shaped or swollen mitochondria were observed in the model group. ST36 treatment improved the disor-dered fiber arrangement, and reduced the population of damaged mitochondria; thus, fused and elongated mitochondria were readily observed. Compared with the model group, there were no obvious improvements in the non-acupoint group. Compared with the normal group, the expression levels of Opa1 and Drp1 mRNAs and proteins in the skeletal muscle tissue were significantly lower in the model group (P<0.05). After the treatment, the expression levels of Opa1 and Drp1 mRNAs and proteins were up-regulated in the ST36 group (P<0.05), and the expression of Drp1 protein was up-regulated in the non-acupoint group (P<0.05)．. CONCLUSION: EA at ST36 can correct the imbalance of mitochondrial fission and fusion in skeletal muscle of rats with spleen qi deficiency, thereby improving the damage of mitochondrial structure and function, and leading to an increase of energy metabolism.

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 formula rapidly. 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.

Untargeted metabonomics study of ginseng in treatment of spleen-Qi deficiency / 中国中药杂志

Ginseng has been used to treat Qi-deficiency syndrome up to now, while the therapeutic mechanism is still unclear. In order to explore the mechanism of ginseng in the treatment of Qi-deficiency constitution, the untargeted metabonomics with blood was studied based on rapid resolution high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(RRLC-Q-TOF-MS). In the results, 13 potential biomarkers were found and identified, which mainly involved in the body's antioxidant and immune functions and energy, glycerol, fatty acid, sugar metabolism and bile acid metabolism. The results of blood biochemical analysis also indicated that ginseng could regulate the body's energy metabolism, immune functions and antioxidant capacity in spleen-Qi deficiency constitution. This study revealed the mechanism of ginseng in the treatment of spleen-Qi deficiency using the blood metabonomics, which could provide technological support and scientific basis for further research on ginseng treatment of Qi-deficiency.

Effects of electroacupuncture at "Zusanli"(ST36) on ultrastructure and mitochondrial dynamics of skeletal muscle in rats with spleen qi deficiency syndrome / 针刺研究

OBJECTIVE: To observe the effects of electroacupuncture (EA) at "Zusanli" (ST36) on the ultrastructure and mitochondrial dynamics of skeletal muscle tissue in spleen qi deficiency rats, so as to explore the partial action mechanism of EA at ST36 for spleen deficiency syndrome. METHODS: Twenty-four male SD rats were randomly divided into 4 groups: normal group, model group, ST36 group and non-acupoint group (n＝6 in each group). The model of spleen qi deficiency syndrome was established by improper diet and exhaustive swimming. EA (2 Hz/15 Hz, 0.5 mA) was applied to bilateral ST36 in the ST36 group and non-acupoint in the non-acupoint group for 20 min, once daily for 7 days. The colorimetric method was used to detect the ATP content in skeletal muscle tissue. The ultrastructure changes of skeletal muscle tissue were observed by transmission electron microscopy. The expression levels of optic atrophy 1 (Opa1) and dynamin-related protein 1 (Drp1) mRNA and proteins in the skeletal muscle tissue were determined by fluorescence quantitative real-time PCR and Western blot, respectively. RESULTS: The ATP content in skeletal muscle tissue of model group was significantly lower than that in the normal group (P<0.05), while significantly higher in the ST36 group than that in the model group and non-acupoint group (P<0.05). Transmission electron microscopy showed that a large number of muscle fibers that were ruptured, damaged, and disorganized; moreover, many vacuoles with different sizes, and abnormally shaped or swollen mitochondria were observed in the model group. ST36 treatment improved the disor-dered fiber arrangement, and reduced the population of damaged mitochondria; thus, fused and elongated mitochondria were readily observed. Compared with the model group, there were no obvious improvements in the non-acupoint group. Compared with the normal group, the expression levels of Opa1 and Drp1 mRNAs and proteins in the skeletal muscle tissue were significantly lower in the model group (P<0.05). After the treatment, the expression levels of Opa1 and Drp1 mRNAs and proteins were up-regulated in the ST36 group (P<0.05), and the expression of Drp1 protein was up-regulated in the non-acupoint group (P<0.05)．. CONCLUSION: EA at ST36 can correct the imbalance of mitochondrial fission and fusion in skeletal muscle of rats with spleen qi deficiency, thereby improving the damage of mitochondrial structure and function, and leading to an increase of energy metabolism.