Guizhi Fuling Research Articles

The crucial component(s) of Guizhi Fuling Wan in inhibiting endometriosis development

Ethnopharmacological relevanceEndometriosis is a common gynecological disorder that manifests as chronic pelvic pain and subfertility. Guizhi Fuling Wan (GFW), which contains five herbs, was first described in Chinese canonical medicine to treat qi stagnation and circulation. Although the inhibition of endometriosis by GFW has been previously demonstrated, its efficacy could potentially be improved by adjusting the dose of each component. AimThis study aimed to examine the relative importance of herbs in endometriosis treatment. Materials and methodsEndometriosis was induced in C57BL/6NCrlBltw mice, followed by treatment with H2O, GFW, all individual herbs of GFW, and GFW with sequential deletion of a single herb for 28 days. Immunohistochemistry and western blotting were performed to examine the expression of inflammatory and apoptotic markers. ResultsThe endometriosis-inhibiting effect of GFW was reduced by deletion of either Guizhi or Fuling. Guizhi, Fuling, or Taoren alone also inhibit the development of endometriosis. The reduction in intercellular adhesion molecule 1 (ICAM-1) by GFW was attenuated by deletion of Guizhi, Fuling, or Taoren, whereas either Guizhi or Fuling alone decreased ICAM-1 expression. The deletion of either Guizhi or Fuling diminished the enhancement of caspase-3 by GFW, whereas caspase-3 expression was elevated by either Guizhi or Fuling alone. ConclusionDeletion of either Guizhi or Fuling attenuated the inhibition of endometriosis development, while either Guizhi or Fuling alone maintained the endometriosis-inhibiting effect of GFW. Consistent with the description in the canonical literature of traditional Chinese medicine, this study revealed that Guizhi and Fuling are the most crucial components of GFW in treating endometriosis, as they are the emperor drugs of this formula, whereas Taoren is a minister drug.

Study on the Mechanism of Guizhi Fuling Decoction in Treating Hyperlipidemia Based on Network Pharmacology and Experimental Methods

Background Hyperlipidemia, caused by abnormal lipid metabolism, has become a significant health concern, especially in younger populations. While statins are commonly used for treatment, they often cause severe side effects with long-term use, leading to increased interest in finding natural alternatives. Purpose This study aimed to investigate how Guizhi Fuling Decoction (GZFL) treats hyperlipidemia using network pharmacology and experimental validation. Materials and Methods Network pharmacology analyzed GZFL’s active components, targets, and treatment mechanisms for hyperlipidemia. Hyperlipidemia rat and high-fat cell models were established. Experimental validation included enzyme-linked immunosorbent assay, Western blot, quantitative polymerase chain reaction, Oil Red O staining, and other methods. Statistical analysis was performed using one-way analysis of variance followed by Tukey’s post hoc test for multiple comparisons. Pearson correlation analysis was used to identify correlations between key protein expressions and lipid levels. Results GZFL comprises 85 active components and targets 218 proteins. Hyperlipidemia involves 3,852 targets, with 175 overlapping targets. Key targets included estrogen receptor 1, prostaglandin-endoperoxide synthase 2, interleukin-6 (IL-6), protein kinase B (AKT) 1, tumor necrosis factor, interleukin-1 beta, peroxisome proliferator-activated receptor gamma (PPARG), tumor protein p53 (TP53), and fructo oligosaccharide (FOS). Major active components were quercetin, phytosterols, kaempferol, and baicalein. Enrichment analysis highlighted processes like lipopolysaccharide response and lipid signaling pathways. Animal studies showed GZFL significantly reduced serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol levels, and increased apolipoprotein A1 ( p < 0.05). Western blot indicated that GZFL influenced AKT, TP53, IL-6, PPARγ expression, and mRNA levels in the phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway ( p < 0.01). Serum pharmacology experiments demonstrated GZFL-medicated serum reduced lipid droplet formation in high-fat cell models. Conclusion GZFL contains multiple pharmacologically active components, such as quercetin, phytosterols, and kaempferol, which can influence lipid metabolism. It may exert its effects by regulating proteins like AKT, TP53, IL-6, and PPARγ, participating in lipid and atherosclerosis signaling pathways and the PI3K/AKT/mTOR pathway.

Research Progress on Pharmacological Effects of Guizhi Fuling Pill

Research Progress on Pharmacological Effects of Guizhi Fuling Pill

Exploring the therapeutic efficacy and pharmacological mechanism of Guizhi Fuling Pill on ischemic stroke: a meta-analysis and network pharmacology analysis.

The role of Guizhi Fuling Pill (GZFL) in the treatment of ischemic stroke (IS) is still controversial, and its pharmacological mechanism remains unclear. To evaluate the efficacy and potential pharmacological mechanisms of GZFL on IS, a comprehensive method integrating meta-analysis, network pharmacology, and molecular docking was employed. Eight electronic databases were searched from inception to November 2023. Review Manager 5.4.1 software was used for meta-analysis. Active compounds and targets of GZFL were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database, Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine, and Encyclopaedia of Traditional Chinese Medicine. Relevant targets of IS were obtained from the DisGeNet, Genecards, and DrugBank databases. GO biological function analysis and KEGG enrichment analysis were performed in the Metascape database. AutoDock Tools and PyMOL software were employed for Molecular docking. The intervention group significantly increased the total effective rate and decreased the NIHSS score. Administration of GZFL also improved the whole blood viscosity (low and high shear rates) and levels of fibrinogen, TNF-α, and IL-6. The key active compounds included quercetin, kaempferol, catechin, and beta-sitosterol, and the core target proteins included SRC, MAPK1, TP53, JUN, RELA, AKT1, and TNF. GO analysis mainly involved inflammation response, cellular response to lipids, and regulation of ion transport. The core pathways were lipid and atherosclerosis, cAMP, calcium, IL-17, and MAPK signaling pathways. Key active compounds showed good affinity with the core targets. The underlying mechanisms of GZFL in IS treatment are primarily related to its anti-inflammatory, anti-atherosclerosis, and neuroprotective effects.

Guizhi Fuling Wan attenuates tetrachloromethane-induced hepatic fibrosis in rats via PTEN/AKT/mTOR signaling pathway

Ethnopharmacological relevanceTreatment options for hepatic fibrosis, a prevalent liver condition closely linked to cirrhosis, are currently limited. While Guizhi Fuling Wan (GFW), a pill derived from traditional Chinese herbs, has been reported to possess hepatoprotective properties, its therapeutic effect and mechanism in hepatic fibrosis remain elusive. Aim of the studyThis study aimed to evaluate the anti-fibrotic impact of GFW and its underlying mechanisms in both in vivo and in vitro settings. Materials and methodsTetrachloromethane (CCl4) was used to induce hepatic fibrosis in male rats. In vitro, activation of hepatic stellate cells (HSCs) was triggered by platelet-derived growth factor-BB (PDGF-BB). In vivo, liver function, pathological alterations, and HSC activation were evaluated. Additionally, the impact of GFW on the activated phenotypes of Lieming Xu-2 (LX-2) cells was examined in vitro. Network pharmacology was employed to identify the potential targets of GFW in hepatic fibrosis. Lastly, the impact of GFW on the PTEN/AKT/mTOR pathway and PTEN ubiquitination in HSCs was investigated. ResultsGFW alleviated CCl4-induced liver damage and scarring in rats in a dose-dependent manner and suppressed HSC activation in vivo. Moreover, GFW inhibited the proliferation, migration, differentiation, and extracellular matrix (ECM) production of activated HSCs in vitro. GFW also promoted autophagy and apoptosis of HSCs. Meanwhile, network pharmacology and in vitro studies suggested that GFW inhibits the AKT/mTOR pathway by preventing PTEN degradation by suppressing ubiquitination. ConclusionGFW attenuates Ccl4-induced hepatic fibrosis in male rats by regulating the PTEN/AKT/mTOR signaling pathway, positioning it as a potential candidate for the treatment of hepatic fibrosis.

Coronary heart disease: innovative understanding from traditional Chinese medicine and treatment by classic formulas

Coronary heart disease is a common cardiovascular disease, attacking about 11.4 million patients in China. With increasing prevalence and mortality year by year, coronary heart disease has become a major factor threatening human health and public health. Although primary and secondary prevention, intervention, coronary artery bypass grafting and other interventions have reduced the death rate, there are drug(aspirin) resistance, secondary nitroglycerin failure, post-intervention fatigue, chest tightness, and an-xiety, and complication with a high risk of bleeding, which have become the key clinical and scientific issues needed to be resolved. Coronary heart disease belongs to the category of chest impediment and heart pain in traditional Chinese medicine(TCM). The TCM etiology of this disease includes external contraction of cold, emotional disorders, constitutional insufficiency, physical weakness, and labor injury, which are closely related to sympathetic nerve activity, state of cardiac and psychological diseases, family history, and cardiovascular metabolic disorders in modern medicine. The TCM causes of coronary heart disease include Qi depression, phlegm turbidity, blood stasis, fire-heat, cold congealing, and healthy Qi deficiency, which are associated with emotional factors such as anxiety and depression, abnormal lipid metabolism, abnormalities in blood circulation and coagulation, inflammatory responses, hyperactive immune responses, and heart failure, chronic wasting disease, or aging, respectively. Accordingly, the patients with Qi depression should be treated with Chaihu Longgu Muli Decoction, and those with phlegm turbidity should be treated with Wendan Decoction and Gualou Xiebai Banxia Decoction. Xuefu Zhuyu Decoction and Guizhi Fuling Pills are recommended for blood stasis, Xiaoxianxiong Decoction and Sanhuang Xiexin Decoction for fire-heat, Zhishi Xiebai Guizhi Decoction for cold congealing, and Renshen Decoction for healthy Qi deficiency. Due to the changes in the spectrum of diseases from ancient to modern times as well as the differences in physical constitution, the key cause of coronary heart disease has evolved from the chest Yang deficiency and cold congealing to Qi depression, phlegm turbidity, phlegm combined with stasis, and fire-heat, showing a shift from cold to heat and from deficiency to excess. The combination of classic formulas presents a pattern. That is, the core formula-syndrome correspondence of a disease often fixedly appears with other formula-syndrome correspondence, which may be related to the development of the pathophysiological mechanism of the disease. In the clinical application of modern pharmacological results, the research team has formulated the clinical principle of pathogenesis corresponding to pathological changes and medicinal nature corresponding pharmacological effects. The modern pharmacological research on classic formulas is conducive to targeted treatment. Moreover, classic formulas help to ameliorate aspirin resistance, clopidogrel resistance, post-intervention anxiety, and high risk of bleeding and address the lack of effective blockade of critical lesions in the coronary artery and the progression of post-infarction heart failure. The innovative understanding of the etiology and pathogenesis of co-ronary heart disease helps to improve the clinical efficacy of TCM and the clinical system for treating coronary heart disease with classic formulas.

Comparative efficacy of commercial oral poly-herbal traditional Chinese medicine formulations combined with western medicine in benign prostatic hyperplasia management: a systematic review and network meta-analysis.

Background: Benign prostatic hyperplasia (BPH) is prevalent among the aging male population and often presents with distressing lower urinary tract symptoms. There is emerging evidence that commercial oral poly-herbal traditional Chinese medicine (TCM) formulation combined with Western medicine (WM) may offer enhanced therapeutic effects compared to WM alone in BPH treatment. Nevertheless, determining the optimal formulations for BPH remains controversial. We aimed to employ a network meta-analysis to compare and assess differences among commonly used and recommended poly-herbal TCM formulations outlined in the Chinese guidelines for BPH treatment, providing clinical medication recommendations and guidance. Methods: We extensively searched for RCTs of BPH patients that had oral poly-herbal TCM formulations and WM treatment, covering both English and Chinese databases up to 31 October 2023. The quality of the included studies was evaluated using the Cochrane risk-of-bias tool Version 2 (ROB2). A Bayesian network meta-analysis was performed to assess the effectiveness of various formulations, followed by sensitivity and subgroup analyses. Results: Our meta-analysis included 107 RCTs involving 11,037 patients across 16 oral poly-herbal TCM formulations. The quality of the selected studies was assessed as "Some concerns". Most formulations combined with WM demonstrated superior therapeutic efficacy compared to WM alone. For clinical effective rate, Jingui Shenqi pill (JGSQ) + WM had the highest-ranking probability (87.38%). Concerning International Prostate Symptom Score (IPSS) and maximum flow rate of urine, Guizhi Fuling capsule (GZFL) + WM was most effective (91.10% and 98.55%). Regarding the quality of life score and postvoid residual urine, Pulean tablet (PLA) + WM ranked first (86.71% and 91.81%). In controlling prostate volume, Huange capsule (HE) + WM demonstrated the highest efficacy (95.65%). Additionally, among the interventions, Lingze (LZ) + WM capsule exhibited the lowest incidence of adverse drug reactions (2.32%). Conclusion: Combining oral poly-herbal TCM formulations with WM may provide greater therapeutic benefits in BPH treatment compared to WM alone. JGSQ, GZFL, PLA, and HE emerged as promising treatment options. However, further rigorous empirical studies are essential to substantiate these findings. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=459651, CRD 42023459651.

Guizhi Fuling Wan ameliorates concanavalin A-induced autoimmune hepatitis in mice

BackgroundAutoimmune hepatitis (AIH) is an immune-mediated hepatic disease associated with intense complications. AIH is more common in females and needs effective drugs to treat. Guizhi Fuling Wan (GZFLW) is a traditional Chinese herbal formula used to treat various gynecologic diseases. In this study, we aim to extend the new use of GZFLW for AIH. MethodsThe tandem MS-based analysis was used to identify secondary metabolites in GZFLW. Therapeutic effects of GZFLW were tested in a concanavalin A (Con A)-induced AIH model in mice. Ethnopharmacological mechanisms underlying the antiapoptotic, antioxidant, and immunomodulatory protective effects were determined. ResultsOral administration of GZFLW attenuates AIH in a Con A-induced hepatotoxic model in vivo. The tandem MS-based analysis identified 15 secondary metabolites in GZFLW. The Con A-induced AIH syndromes, including hepatic apoptosis, inflammation, reactive oxygen species accumulation, function failure, and mortality, were significantly alleviated by GZFLW in mice. Mechanistically, GZFLW restrained the caspase-dependent apoptosis, restored the antioxidant system, and decreased pro-inflammatory cytokine production in the livers of Con A-treated mice. Besides, GZFLW repressed the Con A-induced hepatic infiltration of inflammatory cells, splenic T cell activation, and splenomegaly in mice. ConclusionsOur findings demonstrate the applicable potential of GZFLW in treating AIH. It prompts further investigation of GZFLW as a treatment option for AIH and possibly other hepatic diseases.

Gallic acid ameliorates endometrial hyperplasia through the inhibition of the PI3K/AKT pathway and the down-regulation of cyclin D1 expression

BackgroundGallic acid (GA) is an organic compound with phenolic properties that occurs naturally and can be found in Guizhi Fuling capsules, showcasing a wide range of biological functionalities. PurposeThe objective of this study was to examine the influence of GA on endometrial hyperplasia (EH) and elucidate its underlying mechanism. MethodsInitially, the induction of EH was achieved by administering estradiol to mice via continuous subcutaneous injection for a duration of 21 days. Concurrently, GA treatment was administered, and subsequently, the uterine tissue structure was assessed using hematoxylin and eosin (H&E) staining. Following this, the proliferation of human endometrial cells treated by GA was determined utilizing the CCK-8 method. Furthermore, network pharmacology and single-cell-RNA-seq data were employed to identify the target of GA action. In addition, we will employ immunofluorescence (IF), immunohistochemistry (IHC), flow cytometry, western blot and RT-qPCR methodologies to investigate the impact of GA on the expression level of cyclin D1, PI3K, p-PI3K, AKT, p-AKT. ResultsGA treatment ameliorated histopathological alterations in the uterus and suppress proliferation. Estradiol stimulation can activate the PI3K/AKT pathway, leading to up-regulation of cyclin D1 expression, whereas GA treatment results in down-regulation of its expression. ConclusionsThe expression of cyclin D1 is down-regulated by GA through the inhibition of the PI3K/AKT pathway, effectively mitigating estradiol-induced EH in mice.

Research Progress on the Treatment of Benign Prostatic Hyperplasia with Classical Prescriptions

AbstractThe main pathogenesis of benign prostatic hyperplasia (BPH) includes deficiency of healthy qi, blood stasis, and fluid retention. The root cause of BPH is the deficiency of healthy qi, for which Jingui Shenqi Pill and Zhenwu Decoction are used to nourish kidney qi and warm yang to promote diuresis. Blood stasis is the key pathogeneses of BPH, for which Guizhi Fuling Pill and Didang Decoction are used to dissipate blood stasis, relieve obstruction of orifices and facilitate bowel movement. Fluid retention is the most urgent symptom of BPH, for which Wuling Powder and Danggui Shaoyao Powder are used to transform qi and promote diuresis, addressing the urgent symptoms.

Computer Prediction and Experimental Verification of Guizhi Fuling Pill in the Treatment of Benign Prostatic Hyperplasia

Objective: Benign prostatic hyperplasia (BPH) is a prevalent condition among middle-aged and elderly males. The hypoxia-inducible factor-1α (HIF-1α) / vascular endothelial growth factor A (VEGF-A) signaling pathway plays a key role in proliferative disorders. Guizhi Fuling Pill (GFP) provides significant clinical efficacy in addressing BPH and related hyperplastic ailments. This study aims to elucidate the potential involvement of the HIF-1α / VEGF-A signaling pathway in the therapeutic action of GFP against BPH. Methods: Employing network pharmacology and molecular docking techniques, the mechanism underlying GFP against BPH was anticipated. A rat model of BPH was established to validate the primary targets identified via molecular docking. Subcutaneous administration of testosterone propionate was conducted to induce the BPH rat model for experimental validation. Immunohistochemistry was performed to assess the expression of HIF-1α and VEGF-A in prostate tissue across different experimental groups. Results: The network pharmacology analysis unveiled 111 potential constituents of GFP, with key components such as ent-Epicatechin, quercetin, and Mairin exerting influence on BPH. Essential targets encompass ESR1 and HSP90AA1, with signaling pathways like HIF-1, PI3K-AKT, and VEGF prominently associated with cellular apoptosis, angiogenesis, and proliferation. Molecular docking projections underscored the propensity of core components to interact with key targets, exhibiting favorable binding characteristics. In the context of animal experiments, GFP significantly reduced the expression of HIF-1α and VEGF-A in BPH-afflicted rats as compared to the model group. Conclusion: Collectively, this investigation suggests that GFP’s therapeutic potential for BPH could be attributed to its modulation of multifaceted targets, including HIF-1α and VEGF-A.

The potential mechanism of Guizhi Fuling Wan effect in the treatment of cervical squamous cell carcinoma: A bioinformatics analysis investigation.

As a global malignancy with high mortality rate, targeted drug development for Uterine Cervical Neoplasms is an important direction. The traditional formula Guizhi Fuling Wan (GFW) is widely used in gynecological diseases. However, its potential mechanism of action remains to be discovered. We retrieved GFW and cervical squamous cell carcinoma (CSCC) targets from public databases. The protein-protein interaction network was obtained by string computational analysis and imported Cytoscape_v3.9.0 to obtain the core network and the top 10 Hub genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes were used for enrichment analysis of the core network, and then molecular docking to verify whether the selected signaling pathway binds well to the core node. Finally, clinical prognostic analysis and expression differences of Hub genes were validated using the Cancer Genome Atlas database and R language. Our search yielded 152 common targets for GFW and CSCC. The interleukin-17 signaling pathway, tumor necrosis factor signaling pathway, and Toll-like signaling pathway were then selected for further molecular docking from the hub genes enrichment analysis results, which showed good binding. Among the Hub genes, JUN, VEGFA, IL1B, and EGF had a poor prognosis for CSCC. In conclusion, this study illustrates that GFW can have adjuvant therapeutic effects on CSCC through multiple targets and multiple pathways, providing a basis for further research.

参苓白术散联合桂枝茯苓丸、达格列净治疗2型糖尿病肾病IV期的蛋白尿的随机对照研究

参苓白术散联合桂枝茯苓丸、达格列净治疗2型糖尿病肾病IV期的蛋白尿的随机对照研究

The ameliorating effects of Guizhi Fuling Wan combined with rosiglitazone in a rat ovarian model of polycystic ovary syndrome by the PI3K/AKT/NF-κB and Nrf2/HO-1 pathways

Objective GuizhiFulingWan (GFW) has been reported to be effective against polycystic ovary syndrome (PCOS) by possessing oxidative stress and inflammation which related to PI3K/AKT/NF-κB, Nrf2/HO-1 pathway. This study aims to probe the effects and mechanisms of GFW combined with rosiglitazone on PCOS via PI3K/AKT/NF-κB and Nrf2/HO-1 pathways. Methods A rat PCOS model established by dehydroepiandrosterone (DHEA) injection. The experiment was allocated to control, DHEA, GFW, rosiglitazone, GFW + rosiglitazone groups. Treatment for 30 days, we monitored weight and ovarian weight of rats. Fasting blood glucose (FBG), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), lipid metabolism indexes, estrous cycle and sex hormone-, inflammation-, oxidative stress-related factors were examined. Hematoxylin&eosin staining assessed ovarian tissue pathological changes. Western blot determined PI3K/AKT/NF-κB, Nrf2/HO-1 pathways-related markers. Results GFW and rosiglitazone treatment suppressed body weight and ovarian weight in PCOS rats. They also decreased FBG, FINS, HOMA-IR while inhibited total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL) and enhanced high-density lipoprotein (HDL). They ameliorated estrous cycle, ovarian histological changes and follicular development. They restrained testosterone (T), luteinizing hormone (LH) and accelerated estradiol (E2), progesterone (P), follicle stimulating hormone (FSH). They inhibited glutathione peroxidase (GSH-Px), malondialdehyde (MDA), superoxide dismutase (SOD) in serum while increased GSH-Px, SOD and decrease MDA in ovarian tissues. They reduced C-reactive protein, interleukin-18 (IL-18), tumor necrosis factor-α (TNF-α), IL-6, IL-1β levels. GFW and rosiglitazone co-intervention regulated PI3K/AKT/NF-κB and Nrf2/HO-1 pathways in PCOS rats. Conclusion GFW alleviated ovarian dysfunction in PCOS rats, which may be related to the PI3K/AKT/NF-κB, Nrf2/HO-1 pathways.

Guizhi Fuling Capsule inhibits migration and induces apoptosis of human ovarian cancer cells by regulating the NF-κB signaling pathway

To study the inhibitory effect of Guizhi Fuling Capsule (GFC) on migration of human ovarian cancer cells and explore the possible mechanism. Sixty Wistar rats were randomized into 4 groups for daily gavage of saline or 4, 8, or 16 g/kg GFC suspension for 5 days to prepare blank and low-, medium- and high-dose GFC-medicated sera. Cisplatinresistant ovarian cancer SKOV3/DDP cells were treated with these sera with nuclear factor-κB (NF-κB) inhibitor SN50 as the positive control, and the changes in migration ability and apoptosis of the cells were examined using scratch assay and flow cytometry, respectively; the changes in the mRNA and protein expressions of CDH1, CDH2, caspase 3 and NF- κB were detected using RT-qPCR and Western blotting. ATAC-seq was used to analyze the changes in expressions of CDH1, CDH2, caspase 3 and NF-κB genes in the open chromatin. Treatment with GFC-medicated sera dose-dependently inhibited the migration (P < 0.05), increased apoptosis (P < 0.01), inhibited CDH2 and NF-κB mRNA expression (P < 0.05), and enhanced caspase 3 and CDH1 mRNA expressions (P < 0.01) in SKOV3/DDP cells. The effects of high-dose GFC-medicated serum were comparable to those of SN50 (P>0.05), but its effect for enhancing DH1 protein expression was weaker than that of SN50 (P < 0.01). GFC-medicated sera significantly lowered the expressions of NF-κB and CDH2 and increased CDH1 expression in the open chromatin without obviously affecting caspase 3 expression. GFC- medicated sera inhibits the migration ability of SKOV3/DDP cells possibly by promoting cell apoptosis and caspase 3 and CDH1 expressions, inhibiting CDH2 and NF-κB expressions, and regulating the expressions of NF-κB, CDH2 and CDH1 in the open chromatin.

Discrete element simulation study of mixing process of Guizhi Fuling Capsules: parameter calibration

The mixing process is a critical link in the formation of oral solid preparations of traditional Chinese medicine. This paper took the extract powder of Guizhi Fuling Capsules and Paeonol powder as research objects. The angle of repose, loose packing density, and particle size of the two powders were measured to calibrate discrete element simulation parameters for the mixing process. The discrete element method was used to calibrate the simulated solid density of Paeonol powder and extract powder of Guizhi Fuling Capsules based on the Hertz-Mindlin with JKR V2 contact model and particle scaling. The Plackett-Burman experimental design was used to screen out the critical contact parameters that had a significant effect on the simulation of the angle of repose. The regression model between the critical contact parameters and the simulated angle of repose was established by the Box-Behnken experimental design, and the critical contact parameters of each powder were optimized based on the regression model. The best combination of critical contact parameters of the extract powder of Guizhi Fuling Capsules was found to be 0.51 for particle-particle static friction coefficient, 0.31 for particle-particle rolling friction coefficient, and 0.64 for particle-stainless steel static friction coefficient. For Paeonol powder, the best combination of critical contact parameters was 0.4 for particle-particle static friction coefficient and 0.19 for particle-particle rolling friction coefficient. The best combination of contact parameters between Paeonol powder and extract powder of Guizhi Fuling Capsules was 0.27 for collision recovery coefficient, 0.49 for static friction coefficient, and 0.38 for rolling friction coefficient. The verification results show that the relative error between the simulated value and the measured value of the angle of repose of the two single powders is less than 1%, while the relative error between the simulated value and the measured value of the angle of repose of the mixed powder with a mass ratio of 1∶1 is less than 4%. These research results provide reliable physical property simulation data for the mixed simulation experiment of extract powder of Guizhi Fuling Capsules and Paeonol powder.

Rapid and nondestructive identification of adulterate capsules by NIR spectroscopy combined with chemometrics

This study aims to develop a rapid and non-destructive method to identify counterfeit and substandard drugs, addressing the critical need for better quality control in drug production. According to the reasons for counterfeit products in actual production, the commonly used solid preparation excipients such as HPMC, MCC, Mg-St and Pregelatinized Starch, as well as three chemical drugs with similar efficacy to Guizhi-Fuling (GZFL) Capsule as adulterants, including Aspirin, Ibuprofen and Sinomenine Hydrochloride were selected and designed as adulteration samples with different levels of adulteration. NIR spectra were collected in a non-invasive mode and analyzed by one-class classification methods. The feasibility of using Near-infrared (NIR) spectroscopy as a detection method to qualitatively identify adulterated samples was explored at three packaging levels of powder, intact capsules and capsules in PVC. The differences between the samples were analyzed by NIR spectra comparison, cluster analysis and principal component analysis. The performance of SVM, OCPLS and DD-SIMCA models in dealing with the authentication of genuine and counterfeit products was established and compared. The results show that the spectra contain sample information and the adulterated samples could be discriminated correctly by established models. Moreover, applying appropriate spectral preprocessing methods can further improve the model's performance. In addition, a PLS regression model was developed to predict the adulteration levels of the three packing level samples, which yielded satisfactory results. This study highlights the potential of NIR spectroscopy combined with Chemometrics as a rapid and non-destructive testing analysis method to accurately identify counterfeit and substandard drugs, thereby ensuring drug quality.

Guizhi Fuling capsule relieves memory deficits by inhibition of microglial neuroinflammation through blocking JAK2/STAT3 pathway in presenilin1/2 conditional double knockout mice.

Chronic neuroinflammation has been regarded as an important part of the pathological initiation of Alzheimer's disease (AD), which is associated with the regulation of microglial activation. Preventing microglial activation to inhibit neuroinflammation may become a potential target for the treatment of neurodegenerative diseases. Guizhi Fuling capsule (GZFL) has a strong repression on inflammatory responses. Here, the presenilin1/2 conditional double knockout (PS cDKO) mice, a well-established mouse model of AD, were divided into: WT mice (WT), WT mice+GZFL (WT+GZFL), PS cDKO mice (cDKO), and PS cDKO mice+GZFL (cDKO+GZFL). Mice in the WT+GZFL and cDKO+GZFL group were fed standard chow containing 2000 ppm GZFL for 90 days. After 60 days of GZFL treatment, mice were given to behavioral tests for 30 days in order to explore the effects of GZFL on cognitive and motor function. Then, mice were sacrificed for examining the effects of GZFL on inflammation. Furthermore, primary microglia were obtained from neonatal Sprague-Dawley rats and pretreated with or without GZFL (50 μg/ml) for 1 h in the absence or presence of lipopolysaccharide (LPS) (100 ng/ml) stimulation to speculate whether the underlying mechanism of GZFL's anti-inflammatory potential was closely associated with Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Our findings indicated that GZFL has the ability to alleviate memory deficits in PS cDKO mice, which attributes to the improvement of neuroinflammation by inhibiting microglial activation and the levels of pro-inflammatory mediators. In addition, GZFL could inverse the tau hyperphosphorylation and the lessened expression of synaptic proteins in hippocampus of PS cDKO mice. Furthermore, GZFL prevented LPS-induced neuroinflammatory responses in primary microglia by decreasing the levels of pro-inflammatory mediators. It is noteworthy that therapeutic effects of GZFL on memory impairment are depended on the inhibition of neuroinflammatory responses by the blockage of JAK2/STAT3 signaling pathway. Taken together, GZFL may be an effective compound Chinese medicine for the improvement and postponement of neurodegenerative progression in AD.

Guizhi Fuling Wan inhibits autophagy of granulosa cells in polycystic ovary syndrome mice via H19/miR-29b-3p

Objective To investigate the potential molecular mechanism of traditional Chinese medicine Guizhi Fuling Wan (GZFLW) inhibiting granulosa cells (GCs) autophagy in polycystic ovary syndrome (PCOS). Methods Control GCs and model GCs were cultured and treated with blank serum or GZFLW-containing serum. The levels of H19 and miR-29b-3p in GCs were detected using qRT-PCR, target genes of miR-29b-3p were identified using luciferase assay. The protein expressions of Phosphatase and tensin homolog (PTEN), Matrix Metalloproteinase (MMP)-2, and Bax were measured using western blot. The level of autophagy was detected via MDC staining, the degree of autophagosomes and autophagic polymers was observed using dual fluorescence-tagged mRFP-eGFP-LC3. Results GZFLW intervention reduced the expression of autophagy-related proteins PTEN, MMP-2 and Bax, by upregulating the expression of miR-29b-3p and downregulated the expression of H19 (p < .05 or p < .01). The number of autophagosomes and autophagy polymers was significantly decreased by GZFLW treatment. However, the inhibition of miR-29b-3p and overexpression of H19 induced a significant increase in the number of autophagosomes and autophagic polymers, which attenuated the inhibitory effect of GZFLW on autophagy (p < .05 or p < .01). In addition, inhibition of miR-29b-3p or overexpression of H19 can attenuate the effect of GZFLW on the expression of PTEN, MMP-2 and Bax proteins (p < .05 or p < .01). Conclusion Our study found that GZFLW inhibits autophagy in PCOS GCs via H19/miR-29b-3p pathway.

Add-on effect of the Guizhi Fuling formula for management of reduced fertility potential in women with polycystic ovary syndrome: A systematic review and meta-analysis of randomized controlled trials.

Guizhi Fuling (GZFL) pill, a traditional Chinese herbal formula including Semen Persicae, Ramulus Cinnamomi, Poria, Radix Paeoniae Alba, and Cortex Moutan, has been widely applied in the management of gynecological diseases. To evaluate the add-on effect of the GZFL formula for treating reduced fertility potential in women with polycystic ovary syndrome (PCOS) by conducting a systematic review and meta-analysis. Two reviewers independently searched the PubMed, Embase, Cochrane Library, Wanfang, SinoMed, and CKNI databases until 09/11/2022. Eligible studies were randomized controlled trials (RCTs) of the GZFL formula plus Western medicine versus the Western medicine for treating PCOS. The primary endpoint was the ovulation, pregnancy, and miscarriage rate. The secondary endpoints included the serum follicle-stimulating hormone (FSH), total testosterone, luteinizing hormone (LH), estradiol, and homeostasis model assessment insulin resistance (HOMA-IR). There were 16 RCTs with 1,385 patients identified. The GZFL formula plus Western medicine significantly improved the ovulation rate (risk ratios [RR] 1.24; 95% confidence intervals [CI] 1.15-1.34) and pregnancy rate (RR 1.53; 95% CI 1.38 to 1.69) than the Western medicine alone. Adjuvant treatment with the GZFL formula also significantly decreased the serum FSH (mean difference [MD] -0.48 U/l; 95% CI -0.80 to -0.15), total testosterone (standard mean difference [SMD] -1.07; 95% CI -1.71 to -0.44), LH level (MD -2.19 U/l; 95% CI -3.04 to -1.34), and HOMA-IR (MD -0.47; 95% CI -0.60 to -0.34). However, there was no significant difference in the miscarriage rate (RR 0.89; 95% CI 0.36-2.20) and serum estradiol level (SMD 0.34; 95% CI -0.25 to 0.94) between two groups. The GZFL formula as adjuvant therapy can improve the ovulation and pregnancy rates in women with PCOS. Its beneficial effects may correlate with reducing FSH, total testosterone, and LH and ameliorating insulin resistance. However, more well-designed RCTs with larger samples and multicenter trials are required to confirm the current findings due to uncertainty of the evidence. PROSPERO identifier, CRD42022354530.