Pachymic Acid Research Articles

Lingguizhugan decoction alleviates gestational diabetes mellitus by modulating the PI3K-AKT pathway and oxidative stress: Network pharmacology and experimental evidence.

The Lingguizhugan decoction (LGZGD) is a promising traditional Chinese medicine for the treatment of gestational diabetes mellitus (GDM). However, its bioactive compounds and therapeutic mechanisms remain unknown. The main chemical composition of LGZGD was analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Furthermore, the underlying mechanisms of LGZGD against GDM were elucidated through network pharmacology and molecular docking. The therapeutic efficacy and targets of LGZGD were further confirmed via an in vitro GDM model (high glucose [HG]-treated HTR-8/SVneo cells). Four compounds of LGZGD, namely, cinnamaldehyde, glycyrrhizic acid, 2-atractylenolide, and pachymic acid, were detected. A total of 26 targets for LGZGD treating GDM were obtained, which were mainly involved in oxidative stress and the PI3K-AKT signaling pathway. The protein-protein interaction (PPI) network unveiled that AKT1, TLR4, TP53, and NOS3 were hub therapeutic targets. Molecular docking showed that these targets had strong affinity with key compounds. In vitro experiments confirmed that LGZGD treatment promoted HG-induced cell viability, migration, and invasion ability while inhibited the apoptosis rate and oxidative stress. Mechanically, western blot revealed that LGZGD may protect HG-treated cells by activating the PI3K-AKT pathway and suppressing TLR4 expression. Our study preliminarily explored the mechanism of LGZGD in GDM treatment, providing a scientific basis for the clinical application of LGZGD.

Pachymic acid promotes ferroptosis and inhibits gastric cancer progression by suppressing the PDGFRB-mediated PI3K/Akt pathway

Gastric cancer (GC) is a common malignant tumour with high incidence and mortality rates worldwide. Despite current treatment modalities, including surgical resection and chemotherapy, challenges such as postoperative recurrence, metastasis and drug resistance persist. Therefore, investigating the feasibility and mechanism of traditional Chinese medicine in treating gastric cancer is crucial for discovering new anti-gastric cancer drugs or adjuvant therapies. Pachymic acid (PA) is a natural triterpenoid found in the traditional Chinese medicinal herb Poria cocos (PC) (Schw. Wolf). Recent studies have reported its inhibitory effects on various cancer cells, including liver, cervical, breast and gastric cancer. Our in vitro and in vivo experiments confirmed that PA inhibits the proliferation, migration and invasion of gastric cancer cells. The treatment of gastric cancer cells with various death inhibitors revealed that PA may suppress gastric cancer progression by inducing ferroptosis. Malondialdehyde, Fe2+, reactive oxygen species and glutathione assays were performed to validate the effects of PA on ferroptosis in gastric cancer. High-throughput sequencing combined with analysis of the TCGA database identified PDGFRB as a potential downstream target of PA. In vivo experiments indicated that the PDGFRB overexpression could counteract the antitumour effects of PA, while ferroptosis induced by the PI3K/Akt signalling pathway may play a key role in this process. This study provides initial evidence that PA, through its interaction with PDGFRB, alters the PI3K/Akt signalling pathway, leading to ferroptosis in gastric cancer cells, thus manifesting its antitumour properties. This discovery holds promise for the development of novel therapeutic strategies for gastric cancer patients.

A review on antitumor effect of pachymic acid.

Poria cocos, also known as Jade Ling and Songbai taro, is a dry fungus core for Wolfiporia cocos, which is parasitic on the roots of pine trees. The ancients called it "medicine of four seasons" because of its extensive effect and ability to be combined with many medicines. Pachymic acid (PA) is one of the main biological compounds of Poria cocos. Research has shown that PA has various pharmacological properties, including anti-inflammatory and antioxidant. PA has recently attracted much attention due to its anticancer properties. Researchers have found that PA showed anticancer activity by regulating apoptosis and the cell cycle in vitro and in vivo. Using PA with anticancer drugs, radiotherapy, and biomaterials could also improve the sensitivity of cancer cells and delay the progression of cancer. The purpose of this review was to summarize the anticancer mechanism of PA by referencing the published documents. A review of the collected data indicated that PA had the potential to be developed into an effective anticancer agent.

Bu-Shen-Ning-Xin decoction ameliorates premature ovarian insufficiency by suppressing oxidative stress through rno_circRNA_012284/rno_miR-760–3p/HBEGF pathway

BackgroundPOI (premature ovarian insufficiency) refers to premature and rapid decline of ovarian reserve function in women before the age of 40, which can be manifested as menstrual disorders, endocrine abnormalities and low fertility. Bu-Shen-Ning-Xin decoction (BSNXD) has been found to have therapeutic effects on POI. Nevertheless, how it exerts therapeutic effects remains elusive. PurposeThis research aims to clarify the pharmacological mechanisms of BSNXD. MethodsWe applied Ultra Performance Liquid Chromatography (UPLC) to identify the main components of BSNXD.4-vinylcyclohexene diepoxide(VCD)was used to induce POI models. ELISA detected the serum level of hormones. H&E staining evaluated the morphology of ovarian tissues.CircRNA and mRNA expression profiles in the ovaries of both POI rats and those treated with BSNXD were detected. Then, dysregulated circRNAs and mRNAs that were potentially altered by BSNXD were screened. Network pharmacology analysis was performed to identify drug targets of BSNXD active ingredients. A circRNA-miRNA-mRNA network and an oxidative stress(OS)-related subnetwork were constructed. Expression of rno_circRNA_012284, rno_miR-760–3p, and HBEGF(Heparin-binding epidermal growth factor-like growth factor) was measured by RT-PCR and their binding were verified by dual-luciferase reporter assays. ROS was measured through DCFH-DA fluorescence probes. The HBEGF target was selected for molecular docking with key active ingredients.Surface plasmon resonance(SPR) was applied to verify the binding ability and affinity between components and HBEGF. ResultsUPLC analysis indicated that 6 chemical compounds including berberine, paeoniflorin, morroniside,gallic acid, loganin, baicalin were identified.Elevated FSH and LH levels, suppressed E2 and AMH levels in the serum, and inhibited follicles and corpus luteums in the ovarian tissues of VCD-induced rats were notably reversed by BSNXD.In total, 992 up- and 1135 down-regulated circRNAs, and 205 up- and 243 down-regulated mRNAs were found in POI rat ovaries following BSNXD administration. Furthermore, 198 drug targets of BSNXD were identified. An OS-related and BSNXD-targeted ceRNA subnetwork composed of rno_circRNA_012284/rno_miR-760–3p/HBEGF was established. rno_circRNA_012284 and HBEGF were up-regulated and rno_miR-760–3p was down-regulated in POI ovarian granulosa cells (OGCs) after BSNXD administration. rno_circRNA_012284 was a sponge of rno_miR-760–3p to elevate HBEGF expression. Moreover, rno_circRNA_012284 overexpression alleviated POI-induced excessive ROS generation in ovarian granulosa cells, while rno_circRNA_012284 inhibition exerted the opposite effect. Finally,molecular docking speculated active ingredients of each herb acted on HBEGF to reduce the OS. SPR tests showed that Berberine,Baicalein,Quercetin,Pachymic acid,Paeoniflorin exhibited satisfying affinity with HBEGF protein. ConclusionThis study demonstrates that BSNXD ameliorates POI partly by attenuating OS in ovarian granulosa cells via rno_circRNA_012284/rno_miR-760–3p/HBEGF axis, uncovering the pharmacological mechanisms of BSNXD in alleviating POI.

Drug-drug interaction between pachymic acid and repaglinide in rats and its potential mechanism

Both repaglinide and pachymic acid are useful for reducing blood glucose, but the drug interactions between them are not known. This study aimed to explore the effect of pachymic acid on the pharmacokinetics of repaglinide in rats and to elucidate its mechanism. The effects of pachymic acid (5 and 10 mg/kg) on the pharmacokinetics of repaglinide (0.5 mg/kg) were investigated in Sprague-Dawley rats (n = 6) with a single dose of repaglinide as a control. LC-MS/MS was used to determine the plasma concentration of repaglinide. Furthermore, the effect of pachymic acid on the metabolic stability of repaglinide and CYP3A4 activity was assessed in rat liver microsomes (RLMs). The co-administration of repaglinide and pachymic acid caused considerable changes in the pharmacokinetics of repaglinide, as shown by an increase in Cmax, t1/2, and AUC(0-t), and a decrease in CLz/F. The effect of pachymic acid was enhanced with increasing concentration. In vitro, pachymic acid showed a significant improvement of the metabolic stability of repaglinide in RLMs and inhibited the activity of CYP3A4. Co-administration of repaglinide with pachymic acid increased the systemic exposure of repaglinide and improved its metabolic stability in rats, which was mediated by CYP3A4. The results of animal studies cannot be directly applied to clinical practice, but concomitant administration of repaglinide and pachymic acid should be avoided or monitored until interaction studies are performed in humans.

Optimizing liquid fermentation for Wolfiporia cocos: gene expression and biosynthesis of pachymic acid and mycelial biomass.

Wolfiporia cocos, a versatile fungus acclaimed for its nutritional and therapeutic benefits in Traditional Chinese Medicine, holds immense potential for pharmaceutical and industrial applications. In this study, we aimed to optimize liquid fermentation techniques and culture medium composition to maximize mycelial biomass (MB) yield, pachymic acid (PA) concentration, and overall PA production. Additionally, we investigated the molecular basis of our findings by quantifying the expression levels of genes associated with PA and MB biosynthesis using quantitative real-time polymerase chain reaction. Under the optimized fermentation conditions, significant results were achieved, with maximum MB reaching 6.68 g l-1, PA content peaking at 1.25 mg g-1, and a total PA yield of 4.76 g l-1. Notably, among the four examined genes, squalene monooxygenase, exhibited enhanced expression at 0.06 ratio under the optimized conditions. Furthermore, within the realm of carbohydrate-active enzymes, the glycoside hydrolases 16 family displayed elevated expression levels at 21 ratios, particularly during MB production. This study enhances understanding of genetic mechanism governing MB and PA production in W. cocos, highlighting the roles of squalene monooxygenase and glycoside hydrolases 16 carbohydrate-active enzymes.

An omics-based characterization of Wolfiporia cocos reveals three CYP450 members involved in the biosynthetic pathway of pachymic acid

Wolfiporia cocos is a medicinal mushroom used in China. It biosynthesizes pachymic acid (PA), a main therapeutic triterpene associated with therapies. Nowadays, the unknown PA biosynthesis leads to difficulties in increasing its content in W. cocos. Herein, we report sequencing, assembling, and characterization of the genome and several transcriptomes of W. cocos. Sequence mining determined candidate genes that encode lanosterol synthase, sterol O-acyltransferase, and sterol C-24 methyltransferase likely involved in the steps from lanosterol to PA. Gene cluster analysis identified four CYP450 cDNAs likely involved in the biosynthesis of PA, namely WcCYP64-1, WcCYP64-2, WcCYP52, and WcCYP_FUM15, which were subjected to both overexpression and silencing in mycelia. The overexpression of each of WcCYP64-1, WcCYP52 and WcCYP_FUM15 increased the content of PA, 16α-hydroxytrametenolic acid, eburicoic acid, and tumulosic acid, while the silencing of each gene either significantly or slightly decreased the contents of these four compounds, indicating their involvement in the PA biosynthesis. In addition, different temperatures affected the expression of these genes and the formation of PA. By contrast, the overexpression and silencing of WcCYP64-2 did not alter the formation of these compounds. Taken together, these findings determine more potential steps in the biosynthetic pathway of PA for metabolic engineering.

Pachymic acid activates TP53INP2/TRAF6/caspase-8 pathway to promote apoptosis in renal cell carcinoma cells.

While pachymic acid (PA), a key component of Poria cocos (Schw.), has demonstrated anti-tumor effects in lung, breast, and pancreatic cancers, its impact on renal cell carcinoma (RCC) is unclear. This study evaluated the effect of PA on proliferation, migration, and apoptosis in human renal cancer A498 and ACHN cells as well as in cancer xenograft mice using wound scratch test, Western blotting, and co-immunoprecipitation assays. In a dose- and time-dependent manner, PA exhibited significant inhibition of RCC cell proliferation, migration, and invasion, accompanied by the induction of apoptosis. Additionally, PA upregulated the expression of tumor protein p53-inducible nuclear protein 2 (TP53INP2) and tumor necrosis factor receptor-associated factor 6 (TRAF6), which were downregulated in renal papillary and chromophobe carcinoma, resulting in inhibited tumor growth in mice. PA treatment elevated cleaved-caspase 3 and 8, and PARP levels, and facilitated TP53INP2 and TRAF6 binding to caspase 8, promoting its ubiquitination. Molecular docking revealed interactions between PA and TP53INP2, TRAF6. In summary, PA inhibits RCC development by upregulating TP53INP2 and promoting TRAF6-induced caspase 8 ubiquitination, activating apoptotic pathways.

Pachymic acid exerts antitumor activities by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B

Objective: To determine the inhibitory effects of pachymic acid on lung adenocarcinoma (LUAD) cells and elucidate its underlying mechanism. Methods: CCK-8, wound healing, Transwell, Western blot, tube formation, and immunofluorescence assays were carried out to measure the effects of various concentrations of pachymic acid on LUAD cell proliferation, metastasis, angiogenesis as well as autophagy. Subsequently, molecular docking technology was used to detect the potential targeted binding association between pachymic acid and protein tyrosine phosphatase 1B (PTP1B). Moreover, PTP1B was overexpressed in A549 cells to detect the specific mechanisms of pachymic acid. Results: Pachymic acid suppressed LUAD cell viability, metastasis as well as angiogenesis while inducing cell autophagy. It also targeted PTP1B and lowered PTP1B expression. However, PTP1B overexpression reversed the effects of pachymic acid on metastasis, angiogenesis, and autophagy as well as the expression of Wnt3a and β-catenin in LUAD cells. Conclusions: Pachymic acid inhibits metastasis and angiogenesis, and promotes autophagy in LUAD cells by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B.

Characterisation and evaluation of physical properties of AH-Plus sealer with and without the incorporation of petasin, pachymic acid, curcumin and shilajit-an invitro study

BackgroundAH Plus, an epoxy resin-based sealer, is widely used in endodontic practice, owing to its good physical properties that confers longstanding dimensional stability and good adhesion to dentin. Nevertheless, its propensity to trigger inflammation, especially in its freshly mixed state, has been extensively documented. Phytochemicals such as Petasin, Pachymic acid, Curcumin, and Shilajit are known for their anti-inflammatory and analgesic effects. This study aimed to analyze and determine the effect of these natural products on the physical properties of AH Plus sealer when incorporated with the sealer.MethodsAH Plus (AHR) sealer was mixed with 10% petasin, 0.75% pachymic, 0.5% and 6%shilajit to obtain AHP, AHA, AHC and AHS in the ratio of 10:1 and 5:1 respectively. Five samples of each material were assessed for setting time, solubility, flow, and dimensional stability in accordance with the ISO 6876:2012 standardization. Sealers were characterized through scanning electron microscopy (SEM), X-ray energy dispersive spectroscopy, X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Statistical evaluation involved the Kolmogorov-Smirnov and Shapiro-Wilks tests for normality and the one-way ANOVA test for analysis.ResultsIn this investigation, the characterisation analysis revealed a relatively similar microstructure in all the experimental root canal sealers. All experimental groups, excluding the control group, exhibited an increase in flow ranging from 11.9 to 31.4% at a 10:1 ratio. Similarly, for the 5:1 ratio, the increase ranged from 12.02 to 31.83%. In terms of dimensional stability, all groups at the 10:1 ratio showed a decrease compared to the control group. The addition of natural agents to AHR in 10:1 ratio led to a reduction in setting time by 8.9–31.6%, and at a 5:1 ratio, the reduction ranged from 8.1 to 31.5%. However, regarding solubility, the addition of natural agents did not induce any significant alterations.ConclusionBased on the results of this study, it can be concluded that all tested root canal sealers exhibited properties that met the acceptable criteria outlined in the ISO 6876:2012 standardization.

Integrated analysis and separation of 5-lipoxygenase inhibitors from triterpenes of Poria cocos using bioaffinity ultrafiltration UPLC-Q-Exactive, molecular docking and target-based multiple complex networks

Poria cocos (Schw.) Wolf (P. cocos) has been widely used as medical plant in East Asia with remarkable anti-Alzheimer's disease (anti-AD) activity. However, the underlying mechanisms are still confused. In this study, based on the β-Amyloid deposition hypothesis of AD, an integrated analysis was conducted to screen and separation 5-lipoxygenase (5-LOX) inhibitors from triterpenoids of P. cocos and investigate the anti-AD mechanisms, containing bioaffinity ultrafiltration UPLC-Q-Exactive, molecular docking, and multiple complex networks. Five triterpenoids were identified as potential 5-LOX inhibitors, including Tumulosic acid, Polyporenic acid C, 3-Epi-dehydrotumulosic acid, Pachymic acid and Dehydrotrametenolic acid. Five potential 5-LOX inhibitors were screened by ultrafiltration affinity assay in P. cocos. The molecular docking simulation results are consistent with the ultrafiltration experimental results, which further verifies the accuracy of the experiment. The commercial 5-LOX inhibitor that Zileuton was used as a positive control to evaluate the inhibitory effect of active ingredients on 5-LOX. Subsequently, the established separation method allowed the five active ingredients (Pachymic acid, 3-Epi-dehydrotumulosic acid, Dehydrotrametenolic acid, Tumulosic acid and Polyporenic acid C) with high purity to be isolated. Targeting network pharmacology analysis showed that five active ingredients correspond to a total of 286 targets. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis found that target cells were mainly enriched in Pathways in cancer, Lipid and atherosclerosis. Our results indicate that P. cocos extract has the potential to be used in the prevention and treatment of neurodegenerative diseases. This will help elucidate the mechanisms of action of various medicinal plants at the molecular level and provide more opportunities for the discovery and development of new potential treatments from health food resources.

Mechanism of pachymic acid in the treatment of gastric cancer based on network pharmacology and experimental verification.

Pachymic acid (PA) is derived from Poria cocos. PA has a variety of pharmacological and inhibitory effects on various tumors. However, the mechanism of action of PA in gastric cancer (GC) remains unclear. To investigate the mechanism of PA in treating GC via the combination of network pharmacology and experimental verification. The GeneCards and OMIM databases were used to derive the GC targets, while the Pharm Mapper database provided the PA targets. Utilizing the STRING database, a protein-protein interaction network was constructed and core targets were screened. The analyses of Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set enrichment analysis were conducted, and molecular docking and clinical correlation analyses were performed on the core targets. Ultimately, the network pharmacology findings were validated through in vitro cell assays, encompassing assessments of cell viability, apoptosis, cell cycle, cloning, and western blot analysis. According to network pharmacology analysis, the core targets were screened, and the PI3K/AKT signaling pathway is likely to be the mechanism by which PA effectively treats GC, according to KEGG enrichment analysis. The experimental findings showed that PA could control PI3K/AKT signaling to prevent GC cell proliferation, induce apoptosis, and pause the cell cycle. Network pharmacology demonstrated that PA could treat GC by controlling a variety of signaling pathways and acting on a variety of targets. This has also been supported by in vitro cell studies, which serve as benchmarks for further research.

P-Cymene and pachymic acid alleviate adalimumab-resistance cell line inflammatory response via inhibiting PI3K/AKT/mTOR signaling pathway

Background: In the management of inflammatory bowel disease, the emergence of drug resistance to monoclonal antibodies has come out as a significant impediment to achieving desired therapeutic outcomes. However, extensive evidence has substantiated that the induction of autophagy holds promise in counteracting the development of drug resistance. In this regard, the utilization of traditional Chinese medicine presents a potential avenue for alleviate drug resistance, specifically through Peilan and Fuling. Consequently, the objective of this study aims to investigate the regulatory effects of p-Cymene and pachymic acid on the autophagy signaling pathway, with the ultimate goal of reducing inflammatory response and alleviating drug resistance. Methods: The RAW 264.7 murine macrophage cell line was subjected to induction using varying concentration gradients of adalimumab in order to establish an adalimumab-resistant cell line. The cell growth curve and viability were assessed using the CCK-8 assay. The expression of cytokines was determined through the implementation of ELISA. The autophagy signaling pathway was evaluated using Western blot analysis. Results: p-Cymene and pachymic acid were found to exert an inhibitory effect on the proliferation of the adalimumab-resistant cell line. Additionally, treatment with p-Cymene and pachymic acid resulted in a decrease in the levels of proinflammatory cytokines, namely TNF-α, IL-1β, and IL-6, while concurrently increasing the level of IL-10. p-Cymene and pachymic acid demonstrated the capacity to suppress the expression of autophagy-related proteins and mRNA including PI3K, AKT, and mTOR. Conclusions: p-Cymene and pachymic acid exhibit the potential to alleviate adalimumab-resistance and ameliorate the inflammatory response, primarily through the activation of the PI3K/AKT/mTOR signaling pathway. These findings suggest that traditional Chinese medicine holds promise as a viable treatment option for mitigating monoclonal antibody resistance in inflammatory bowel disease.

Pachymic Acid Alleviates Oxidative Damage of Retinal Cells Induced by Sodium Iodate or Hydrogen Peroxide by Activating the Nrf2/HO-1 Signaling Pathway

Background: Pachymic acid (PA), extracted from the traditional Chinese medicinal herb Poria cocos ( P cocos) is reported to protect organs such as the liver and lungs from damage. This study investigated the effect of PA on oxidative stress in the retina in vivo and in vitro. Methods: In vitro, retinal pigment epithelial (RPE) cells were treated with hydrogen peroxide to establish a cell model of oxidative damage. The viability of ARPE-19 cells was measured using an MTT kit. Apoptosis of the cells was detected using flow cytometry, and the expression of Caspase-3, Bax, and Bcl-2 was determined. The oxidative stress level was evaluated by observing the production of superoxide dismutase, catalase, and GSH-Px using ELISA in mice serum. The retinal oxidative damage model was induced by intravenous injection of sodium iodate (SI) into the tail vein of mice. OCT, HE, and TUNEL staining monitored the structure and the apoptosis of the mouse retina. The regulatory oxidative factors nuclear transcription factor E2-related factor 2 (Nrf2) and HO-1 were determined using fluorescence staining. Results: H2O2 decreased the viability of RPE cells, which was recovered after PA administration and was shown to reduce the cell apoptosis rate, the expression of Bax, caspase-3, and the production of ROS. In vivo, the thinning of the retina and the apoptosis rate in the retinal tissue of mice caused by the administration of SI were reversed by treatment with PA. Furthermore, PA administration caused the translocation of Nrf2 and increased the expression of HO-1, and the application of these inhibitors inhibited this effect. Conclusion: PA protects RPE cells from oxidative stress and apoptosis by activating the Nrf2/HO-1 pathway, and it prevents retinal damage by halting the progression of retinal thinning in a mouse model of retinal oxidative damage.

Spectrum-effect relationship analysis based on HPLC-FT-ICR-MS and multivariate statistical analysis to reveal the pharmacodynamic substances of Ling-Gui-Zhu-Gan decoction on Alzheimer's disease

Alzheimer’s disease (AD) threatens elderly human health and still lacks effective treatment. Our previous work showed that LGZGD possessed a neuroprotective effect on the Aβ25–35-induced neurotoxicity in differentiated PC12 cells, indicating that LGZGD may be a potential drug for treatment of AD. However, its pharmacodynamic substances which show anti-inflammatory and anti-oxidant stress activities are still unrevealed. This research aims to reveal the pharmacodynamic substances of LGZGD on Aβ25–35-induced PC12 cell model of AD based on a spectrum-effect relationship study by using HPLC-FT-ICR-MS method and multivariate statistical analysis. Firstly, the chemical composition spectra of different combinations of LGZGD were recorded by HPLC-FT-ICR MS. Subsequently, Aβ25–35-induced PC12 cell model of AD was established and pharmacodynamic experiments were conducted to evaluate their anti-inflammatory and anti-oxidant activities, respectively. Finally, the potential pharmacodynamic substances were screened out through spectrum-effect relationship study accompanied by multivariate statistical analysis including bivariate correlation analysis (BCA), grey relational analysis (GRA), principal component analysis (PCA), partial least squares regression analysis (PLSR). As a result, a total of 96 chemical consistents in different combinations of LGZGD were discovered. Among them, 7 components such as isoglabrolide, licorice saponin E2, licorice saponin N2 and licoisoflavanone were directly linked with the anti-inflammatory effects, and 14 constituents such as tumulosic acid, polyporenic acid C, dehydrotumulosic acid, dehydropachymic acid, and pachymic acid were directly correlated with the anti-oxidative stress activities. In conclusion, we combined the HPLC-FT-ICR-MS spectra with pharmacodynamic indicators to develop the spectrum-effect relationships of LGZGD for the first time, and successfully revealed its potential pharmacodynamic substances in the treatment of AD from the anti-inflammatory and antioxidant pathways in the cell model.

Efficacy of Poria Cocos Extract on Sleep Quality Enhancement: A Clinical Perspective with Implications for Functional Foods.

Since the outbreak of the pandemic started, an increase in the number of sleep disorders, including insomnia and poor sleep quality, has been seen. The pattern will probably continue. This study focuses on the preparation and clinical testing of Poria cocos extract in treating suboptimal sleep quality. The optimal extraction method utilized a 75% ethanol concentration, and the clinical investigation involved subjects with defined poor sleep taking 800 mg of the extract nightly, assessed using the Sleep Questionnaire and polysomnography. The non-parametric Wilcoxon signed-rank test was used for statistical analysis due to the non-normal distribution of the collected data. The study involved 21 insomnia sufferers with a mean age of 55 who were administered Poria cocos extracts. The findings of the study indicate a statistically significant rise in the overall duration of sleep (from 327.395 ± 43.2 min to 356.516 ± 63.21 min, p = 0.014). Additionally, there was a notable decrease in the level of arousal during sleep (from 76.316 ± 44.78 min to 47.989 ± 42.38 min, p = 0.009), and an improvement in the sleep severity index of the sleep questionnaire test. Poria cocos as a natural substance could improve quality of sleep, based on the findings. The study investigates Pachymic acid, a substance found in Poria cocos, as a potential indicator for the development of sleeping aids.

Pachymic acid (PA) inhibits ferroptosis of cardiomyocytes via activation of the AMPK in mice with ischemia/reperfusion-induced myocardial injury.

Pachymic acid (PA) is a lanostane-type triterpenoid with various pharmacological effects. However, little is known about the effect of PA on myocardial infarction (MI) induced by ischemia/reperfusion (I/R). In this study, we aimed to investigate the protective effect of PA and its underlying mechanism. A cellular MI model was established by oxygen-glucose deprivation and reperfusion (OGD/R) treatment in HL-1 cardiomyocytes, and we found that OGD/R treatment decreased cell viability and glutathione peroxide (GSH-Px) activity, increased Fe2+ concentration and lactate dehydrogenase (LDH) activity, promoted malondialdehyde (MDA) and reactive oxygen species (ROS) production, and inhibited the expression of ferroptosis marker proteins SLC7A11 and GPX4 in a time-dependent manner. OGD/R-induced HL-1 cells were pretreated with different concentrations of PA (0, 20, 40, 60 μg/mL) for 24 h, and toxicological experiments showed that 150 μg/mL PA decreased cell viability, while low concentrations of PA had no toxic effect on cells. 20 μg/mL PA reversed the inhibitory effect of OGD/R on cell viability, reduced MDA and ROS production, and Fe2+ accumulation, increased GSH-Px activity and the expression of SLC7A11 and GPX4, and decreased LDH activity, especially at 60 μg/mL PA. Meanwhile, PA promoted the phosphorylation of IRS-1, AKT, and AMPK proteins in a dose-dependent manner. AICAR, an AMPK activator, inhibited ferroptosis, while STO-609, an AMPK inhibitor, largely abolished the effect of PA on OGD/R-induced ferroptosis of HL-1 cells. In addition, PA inhibited ferroptosis and myocardial I/R injury in wild-type mice and AMPK knockout (AMPK-/- ) mice. Collectively, PA inhibited ferroptosis of cardiomyocytes through activating of the AMPK pathway, thereby alleviating myocardial I/R injury in mice.

Xie Zhuo Tiao Zhi formula modulates intestinal microbiota and liver purine metabolism to suppress hepatic steatosis and pyroptosis in NAFLD therapy

BackgroundCurrent evidence indicates a rising global prevalence of Non-Alcoholic Fatty Liver Disease (NAFLD), which is closely associated to conditions such as obesity, dyslipidemia, insulin resistance, and metabolic syndrome. The relationship between the gut microbiome and metabolites in NAFLD is gaining attention understanding the pathogenesis and progression of dysregulated lipid metabolism and inflammation. The Xie Zhuo Tiao Zhi (XZTZ) decoction has been employed in clinical practice for alleviating hyperlipidemia and symptoms related to metabolic disorders. However, the pharmacological mechanisms underlying the effects of XZTZ remain to be elucidated. PurposeThe objective of this study was to examine the pharmacological mechanisms underlying the hypolipidemic and anti-inflammatory effects of XZTZ decoction in a mouse model of NAFLD, as well as the effects of supplementing exogenous metabolites on PO induced cell damage and lipid accumulation in cultured hepatocytes. MethodsA high-fat diet (HFD) mouse model was established to examine the effects of XZTZ through oral gavage. The general condition of mice and the protective effect of XZTZ on liver injury were evaluated using histological and biochemical methods. Hematoxylin and eosin staining (H&E) staining and oil red O staining were performed to assess inflammatory and lipid accumulation detection, and cytokine levels were quantitatively analyzed. Additionally, the study included full-length 16S rRNA sequencing, liver transcriptome analysis, and non-targeted metabolomics analysis to investigate the relationship among intestinal microbiome, liver metabolic function, and XZTZ decoction. ResultsXZTZ had a significant impact on the microbial community structure in NAFLD mice. Notably, the abundance of Ileibacterium valens, which was significantly enriched by XZTZ, exhibited a negative correlation with liver injury biomarkers such as, alanine transaminase (ALT) and aspartate transaminase (AST) activity. Moreover, treatment with XZTZ led to a significant enrichment of the purine metabolism pathway in liver tissue metabolites, with inosine, a purine metabolite, showing a significant positive correlation with the abundance of I. valens. XZTZ and inosine also significantly enhanced fatty acid β-oxidation, which led to a reduction in the expression of pro-inflammatory cytokines and the inhibition of liver pyroptosis. These effects contributed to the mitigation of liver injury and hepatocyte damage, both in vivo and vitro. Furthermore, the utilization of HPLC fingerprints and UPLC-Q-TOF-MS elucidated the principal constituents within the XZTZ decoction, including naringin, neohesperidin, atractylenolide III, 23-o-Acetylalisol B, pachymic acid, and ursolic acid which are likely responsible for its therapeutic efficacy. Further investigations are imperative to fully uncover and validate the pharmacodynamic mechanisms underlying these observations. ConclusionThe administration of XZTZ decoction demonstrates a protective effect on the livers of NAFLD mice by inhibiting lipid accumulation and reducing hepatocyte inflammatory damage. This protective effect is mediated by the upregulation of I.valens abundance in the intestine, highlighting the importance of the gut-liver axis. Furthermore, the presesnce of inosine, adenosine, and their derivatives are important in promoting the protective effects of XZTZ. Furthermore, the in vitro approaching, we provide hitherto undocumented evidence indicating that the inosine significantly improves lipid accumulation, inflammatory damage, and pyroptosis in AML12 cells incubated with free fatty acids.

Anti-bladder cancer and COVID-19 potentials of pachymic acid: clinical features and therapeutic targets

ABSTRACT Pachymic acid, as one of bioactive ingredients in Poria Cocos, has been evidenced preclinically with pharmacological benefits including anti-bladder cancer action. Additionally, pachymic acid has been initiatorily identified in Chinese prescriptions applied for coronavirus disease 2019 (COVID-19) treatment. However, the molecular mechanisms and pharmacological targets associated with mitochondrial functions in pachymic acid against bladder cancer and COVID-19 have not been explored. In this study, network pharmacology and molecular docking detections were employed to identify common, candidate, and core targets that were related to pachymic acid treating bladder cancer COVID-19. Enrichment determinations using core target genes were used to reveal the anti-bladder cancer and COVID-19 molecular mechanisms exerted by pachymic acid. Firstly, all 1150 differentially expressed genes among bladder cancer and COVID-19 were ascertained and a risk score with key genes characterized by different risk groups was established. Furthermore, current study highlighted pachymic acid as a potential compound for anti-bladder cancer and COVID-19. Network pharmacology approaches ascertained ten key targets for pachymic acid against bladder cancer and COVID-19. Molecular docking findings showed that pachymic acid presented effectively binding abilities to key target proteins of chemokine (CC-motif) ligand 2 (CCL2), thrombospondin-1 (THBS1), matrix metallopeptidase 1 (MMP1) in severe acute respiratory syndrome coronavirus 2 ;(SARS-CoV-2). Our study indicates that pachymic acid may be used for treatment of bladder cancer and COVID-19 through increasing tumor immunity and inhibiting SARS-CoV-2 replication based on present bioinformatics findings.

Pachymic Acid Protects Against Bleomycin-Induced Pulmonary Fibrosis by Suppressing Fibrotic, Inflammatory, and Oxidative Stress Pathways in Mice.

Pachymic acid (PA), a natural extract from Poria cocos (Schw.) Wolf, possesses anti-inflammatory and anti-oxidative properties. However, it is still unknown whether PA can protect against bleomycin (BLM)-induced pulmonary fibrosis (PF). In this study, we investigated the effects of PA in mice administered BLM. Our results showed that PA significantly improved lung damage and pathological manifestations. Additionally, PA reduced the levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α, while increasing the level of IL-10. PA also decreased the levels of hydroxyproline and malondialdehyde, and increased the activities of superoxide dismutase and glutathione peroxidase in lung tissue. Furthermore, PA inhibited the increases in pyrin domain-containing protein 3 (NLRP3), ASC, IL-1β, P20, and TXNIP induced by BLM. In conclusion, our study demonstrated the protective effects of PA against BLM-induced PF in mice by suppressing fibrotic, inflammatory, and oxidative stress pathways.