Bioactive constituents and action mechanism of Xiaoyao San for treatment of non-alcoholic fatty liver disease.

Xiaoyao San (XYS) is a classic Chinese medicine prescription. It is traditionally used to relieve syndrome of “liver stagnation and spleen deficiency”, a common syndrome type in traditional Chinese medicine, through soothing liver, tonifying spleen, and nourishing blood. Correspondingly, XYS has long application in the treatment of depression, dyspepsia and liver diseases. Given the rising of cutting-edge researches on XYS, there’s a significant need to diligently uncover the bioactive constituents and action mechanisms of XYS for treating non-alcoholic fatty liver disease accordingly.

Xiaoyao san (XYS) plays an important role in treatment of non-alcoholic fatty liver disease (NAFLD) with liver stagnation and spleen deficiency, but its specific mechanism is still unclear. This study aimed to investigate the material basis and mechanism by means of network pharmacology, metabolomics, systems biology and molecular docking methods. On this basis, NAFLD rat model with liver stagnation and spleen deficiency was constructed and XYS was used to intervene, and liver histopathology, biochemical detection, enzyme-linked immunosorbent assay, quantitative PCR assay and western blotting were used to further verify the mechanism. Through the above research methods, network pharmacology study showed that there were 94 targets in total for XYS in the treatment of NAFLD. Metabolomics study showed that NAFLD with liver depression and spleen deficiency had a total of 73 differential metabolites. Systems biology found that PTGS2 and PPARG were the core targets; Quercetin, kaempferol, naringenin, beta-sitosterol and stigmasterol were the core active components; AA, cAMP were the core metabolites. And molecular docking showed that the core active components can act well on the key targets. Animal experiments showed that XYS could improve liver histopathology, increase 5HT and NA, decrease INS and FBG, improve blood lipids and liver function, decrease AA, increase cAMP, down-regulate PTGS2, up-regulate PPARG, and decrease PGE2 and 15d-PGJ2. In conclusion, XYS might treat NAFLD with liver depression and spleen deficiency by down-regulating PTGS2, up-regulating PPARG, reducing AA content, increasing cAMP, improving insulin resistance, affecting glucose and lipid metabolism, inhibiting oxidative stress and inflammatory response. Communicated by Ramaswamy H. Sarma

This study aimed to explore the mechanism of Xiaoyao San(XYS) in the treatment of three diseases of liver depression and spleen deficiency, ie, depression, breast hyperplasia, and functional dyspepsia, and to provide a theoretical basis for the interpretation of the scientific connotation of "treating different diseases with the same method" of traditional Chinese medicines. Traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP) was used to screen the active components of XYS which underwent principal component analysis(PCA) with the available drugs for these three diseases to determine the corresponding biological activities. The targets of XYS on depression, breast hyperplasia, and functional dyspepsia were obtained from GeneCards, TTD, CTD, and DrugBank databases. Cytoscape was used to plot the "individual herbal medicine-active components-potential targets" network. The resulting key targets were subjected to Kyoto encyclopedia of genes and genomes(KEGG) pathway analysis and gene ontology(GO) enrichment analysis. A total of 121 active components of XYS and 38 common targets in the treatment of depression, breast hyperplasia, and functional dyspepsia were collected. The key biological pathways were identified, including advanced glycation and products(AGEs)-receptor for advanced glycation and products(RAGE) signaling pathway in diabetic complications, HIF-1 signaling pathway, and cancer-related pathways. The key targets of XYS in the treatment of depression, breast hyperplasia, and functional dyspepsia included IL6, IL4, and TNF, and the key components were kaempferol, quercetin, aloe-emodin, etc. As revealed by the molecular docking, a strong affinity was observed between the key components and the key targets, which confirmed the results. The therapeutic efficacy of XYS in the treatment of diseases of liver depression and spleen deficiency was presumedly achieved by reducing the inflammatory reactions. The current findings are expected to provide novel research ideas and approaches to classify the scientific connotation of "treating different diseases with the same method" of Chinese medicines, as well as a theoretical basis for understanding the mechanism of XYS and exploring its clinical applications.

Nonalcoholic Fatty Liver Disease (NAFLD) is characterized by excessive lipid accumulation in hepatocytes and is closely associated with metabolic disturbances such as obesity, dyslipidemia, and insulin resistance. Despite its increasing prevalence and potential progression to severe liver conditions, there is currently no approved pharmaceutical intervention for NAFLD. Traditional Chinese Medicine (TCM) formulations, such as Xiaoyao San (XYS), have shown therapeutic efficacy in treating NAFLD, but the underlying mechanisms remain unclear. This study employed a multi-omics approach to elucidate the therapeutic mechanisms of XYS in NAFLD. A rat model of NAFLD was established using a high-fat diet (HFD). The chemical constituents of XYS were analyzed using UPLC-MS/MS. Transcriptomics and proteomics analyses were performed to identify potential biological targets and signaling pathways involved in the therapeutic effects of XYS. The results were validated using ELISA and Western blotting. UPLC-MS/MS identified 225 prototype chemical components of XYS in the blood. XYS significantly reduced body weight, liver index, and Lee’s index in NAFLD model rats. It ameliorated HFD-induced hepatic steatosis, down-regulated serum levels of ALT, AST, GGT, TG, TC, LDL-C, FBG, IL-1β, IL-6, TNF-α, and ROS, and up-regulated HDL-C levels. Transcriptomics and proteomics analyses revealed that XYS modulated key signaling pathways, including cAMP, TGF-β, NF-κB, and necroptosis. Specifically, XYS down-regulated the expressions of NF-κB, p-NF-κB, FOXO1, TGF-β1, RIP3, and p-MLKL, while up-regulating cAMP, PKA, p-PKA, and PPARα. XYS improves NAFLD by regulating the cAMP/PKA-mediated PPARα, FOXO1, and NF-κB signaling pathways. This study provides a comprehensive understanding of the molecular mechanisms underlying the therapeutic effects of XYS in NAFLD and supports its potential as a novel therapeutic intervention for this condition.

Non-alcoholic fatty liver disease (NAFLD) and depression represent two highly prevalent medical conditions often presenting concurrently. However, currently available pharmacological treatments rarely target both disorders simultaneously. Xiaoyao San (XYS), a traditional herbal formulation from Chinese medicine, has been clinically effective in addressing NAFLD and depressive symptoms, although its precise therapeutic mechanisms remain poorly understood. This study aimed to investigate the underlying mechanisms of XYS in managing comorbid NAFLD and depression by employing network pharmacology (NP) and multi-omics methodologies. A chronic restraint stress (CRS)-induced NAFLD rat model was constructed to simulate the disease conditions. The efficacy of XYS treatment was evaluated through behavioral assessments, histopathological liver examination, biochemical analyses, and measurements of inflammatory and oxidative stress (OS) biomarkers. Further, NP combined with metabolomic and proteomic techniques was applied to uncover pivotal targets and pathways, with subsequent validation via Western blotting (WB). XYS improved depression-like behaviors, reduced hepatic lipid deposition, and lowered levels of hepatic enzymes, blood lipids, and blood glucose. It also attenuated inflammatory and OS responses. Multi-omics analysis identified the PPARα/HMOX1/GPX4 axis as a key target. XYS modulated this pathway to inhibit ferroptosis and reduce hepatic iron accumulation. Specifically, XYS upregulated PPARα and GPX4 expression, downregulated HMOX1 expression, and decreased hepatic iron levels, alleviating both NAFLD and depression. XYS exerts therapeutic effects on NAFLD and depression through multi-target regulation, primarily by modulating the PPARα/HMOX1/GPX4 axis to inhibit ferroptosis. This study provides a novel mechanistic basis for the dual therapeutic efficacy of XYS.

Network pharmacology-guided lipidomics and molecular docking reveals the involvement of sphingolipid metabolism and HIF-1 signaling in Xiaoyao San's blood-nourishing mechanism against depression.

Introduction: The effectiveness and safety of the Chinese herbal medicine (CHM) Xiao Yao San (XYS) used for treating anxiety disorders are still unknown. Thus, we conducted this systematic review with meta-analysis and trial sequential analysis (TSA) to determine its safety and efficacy. Methods: We searched 12 databases for relevant studies from the inception of each database till 10 August 2023. We selected randomized controlled trials to compare the efficacy and safety of XYS (including XYS only and XYS + anxiolytics) to those of anxiolytics in patients with anxiety. Results: We found 14 trials with 1,256 patients in total that met the requirements for inclusion. We assessed the majority of studies (8 out of 14) as being at high risk of bias; 6 were assessed as having a moderate risk of bias. Three trials compared oral XYS to anxiolytic medication, and 11 trials compared oral XYS plus anxiolytics to anxiolytic treatment alone. The pooled results showed that the efficacy of treatment in the XYS + anxiolytics groups was significantly higher than that of the anxiolytics alone group (RR = 1.19; 95% CI: [1.13, 1.26]; p < 0.00001; I2 = 0) and the adverse event rates in the XYS + anxiolytics groups were significantly lower than those in the anxiolytics alone group (RR = 0.44; 95% CI: [0.28, 0.82]; p = 0.001 < 0.05; I2 = 13). The efficacy of treatment in the XYS alone groups was also significantly higher than that of the anxiolytics alone groups (RR = 5.41; 95% CI: [2.23, 13.11]; p < 0.0001; I2 = 0). However, there was no statistical difference between the adverse events of the XYS alone group and the anxiolytics alone group, although the incidence of adverse events in the XYS alone group was lower than that in the anxiolytics alone group. The results of the TSA confirmed the above findings. Conclusion: The use of XYS combined with anxiolytics for treating anxiety was found to be safe and effective. However, although XYS alone is effective in the treatment of anxiety disorder, more large-scale research is needed to investigate adverse events. Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=350358, identifier CRD42022350358.

Serum metabolomics reveals compatibility rules of the antidepressant effects of Xiaoyaosan and its efficacy groups

Objective To discuss the clinical efficacy of Sini powder and scattered seven Atractylodes in the treatment of functional dyspepsia with hepatic stagnation and spleen deficiency syndrome. Methods A total of 100 cases of functional dyspepsia with hepatic stagnation and spleen deficiency syndrome in our hospital were selected and randomly divided into the western medicine group and Chinese medicine group by envelope grouping method.The patients in western medicine group were treated with mosapride tablets, 5mg each time, and the patients in Chinese medicine group were treated with Sini powder and scattered seven Atractylodes, 150mL each time, two times a day for 4 weeks.The efficacy, abdominal distension scores and adverse drug reactions were compared between the two groups. Results In the Chinese medicine group, 40 cases were obviously improved, 8 cases were improved, 2 cases were invalid, and the total clinical effective rate was 96.0%.In the western medicine group, 30 cases were obviously improved, 12 cases were improved and 8 cases were invalid.The total clinical effective rate was 80.4%.The total effective rate of the Chinese medicine group was higher than that of the western medicine group, the difference was statistically significant(χ2=8.239, P=0.041). The abdominal distension scores of the two groups after treatment was lower than those before treatment, but the abdominal distension score of the Chinese medicine group[(2.4±2.7)points]was significantly lower than that of the western medicine group[(2.9±2.2)points], the clinical efficacy of the traditional Chinese medicine group was better than that of the Western medicine group(t=3.794, P=0.006). No serious adverse reactions were found in the two groups. Conclusion Sini powder and scattered seven Atractylodes can effectively improve the clinical symptoms of liver stagnation and spleen deficiency in patients with functional dyspepsia, and has no obvious adverse reactions. Key words: Functional dyspepsia; Sini powder and scattered seven Atractylodes; Liver stagnation and spleen deficiency

Xiaoyao San, a Chinese herbal formula, ameliorates depression-like behavior in mice through the AdipoR1/AMPK/ACC pathway in hypothalamus

Xiaoyao San ameliorates high-fat diet-induced anxiety and depression via regulating gut microbiota in mice

Background:Patients with long-term insomnia generally experience anxiety and depression. Traditional sleeping pills and anti-anxiety drugs have certain limitations. Xiao Yao San (XYS), a complementary and alternative therapy, has been widely used to treat insomnia combined with anxiety. This study aims to evaluate the efficacy and safety of XYS in the treatment of insomnia combined with anxiety.Methods:Related literature was retrieved from 8 electronic databases from the establishment time to March 2021. The subjects were diagnosed with insomnia combined with anxiety. We assessed the methodological quality of randomized controlled trials (RCTs) according to the Cochrane Handbook. Data analysis was conducted using RevMan 5.3 software.Results:The analysis includes 9 RCTs involving 681 patients. Meta-analysis supported that as an auxiliary drug for Western medicine (WM), XYS could enhance the clinical efficacy of insomnia calculated according to the traditional Chinese medicine (TCM) syndrome score scale (risk ratio [RR]: 1.26; 95% confidence interval [CI]: 1.13–1.43; P = .0002) and reduced the scores of Hamilton Anxiety Scale (mean difference [MD]: −5.19; 95% CI: −7.78 to −2.60; P < .001), Pittsburgh Sleep Quality Index (MD: −3.35; 95% CI: −4.86 to −1.84; P < .001), Self-rating Anxiety Scale (MD: −9.38; 95% CI: −10.20 to −8.75; P < .001), TCM syndrome score scale for insomnia (MD: −4.45; 95% CI: −6.65 to −2.24; P < .001), and TCM syndrome score scale for anxiety (MD: −5.54; 95% CI: −9.48 to −1.6; P = .006). The summary analysis also shows that the separate use of XYS was advantageous in reducing the scores of the Hamilton Anxiety Scale (MD: −3.70; 95% CI: −6.31 to −1.09; P = .005), Pittsburgh Sleep Quality Index (MD: −1.82; 95% CI: −2.39 to −1.24; P < .001), and Self-rating Anxiety Scale (MD: −10.79; 95% CI: −14.09 to −7.49; P < .001). The incidence of adverse events with XYS as an ancillary drug or used separately was lower than that in the WM.Conclusion:Our systematic evaluation and meta-analysis showed that XYS combined with WM or XYS alone was beneficial for improving sleep quality and relieving anxiety. Due to the low methodological quality, small sample size, and significant heterogeneity of RCTs, more rigorous and scientific clinical trials are required to further evaluate the efficacy and safety of XYS.PROSPERO registration number:CRD42020190613.

Relationship between the phenotypes and functions of peripheral blood dendritic cells and the different spleen deficiency syndrome types in patients with chronic hepatitis B

Efficacy and safety of Chinese patent medicine Xiao Yao San in polycystic ovary syndrome: A systematic review and meta-analysis

Studies on the potential link between antidepressant effect of Xiaoyao San and its pharmacological activity of hepatoprotection based on multi-platform metabolomics.