Atractylodes macrocephala compounds attenuate pediatric epilepsy neuroinflammation through multitarget regulation of the NF-κB pathway

Atractylenolide III attenuates acute kidney injury through phosphorylation of PIK3CA: Functional activation and molecular interaction analysis.

Atractylenolide I (AO-I) and atractylenolide III (AO-III) are the major sesquiterpenes considered to be bioactive from Atractylodes macrocephala (RAM) and its different processed products, popularly used for the treatment of the various digestive diseases and tumors. A rapid and simple capillary zone electrophoresis (CZE) method to quantitatively determine the AO-I and AO-III is described. Using an optimized method, AO-I and AO-III were separated in less than 10 min with A 50 mM borate buffer and a separated voltage 20 kV at 25°C. The CZE method was validated for linearity, sensitivity, accuracy, and precision and then used to determine the content of AO-I and AO-III. This method was successfully applied to determine AO-I and AO-III in real sample RAM and its different processed products.

Atractylenolide III from Atractylodes macrocephala Koidz promotes the activation of brown and white adipose tissue through SIRT1/PGC-1α signaling pathway

Atractylenolide-III restrains cardiac fibrosis after myocardial infarction via suppression of the RhoA/ROCK1 and ERK1/2 pathway.

A simple and sensitive capillary gas chromatography-mass spectrometry (GC–MS) method for the simultaneous determination of atractylenolide I (AO-I) and atractylenolide III (AO-III) in atractylodes macrocephala is described. AO-I and AO-III were obtained by supercritical fluid extraction, and imperatorin was used in this method as an internal standard for quantification. MS detection was performed in SIM mode to increase the sensitivity. Stability of the solutions, linear concentration range, accuracy, precision, LOD, LOQ, and specificity were examined to test the reliability of this method. The method was applied to the analysis of AO-I and AO-III in atractylodes macrocephala.

Atractylenolide-III (AT-III) is well known as its role in antioxidant and anti-inflammatory. Present study was aimed to figure out its effects on osteoarthritis and potential mechanisms. Rat model, human osteoarthritis cartilage explants as well as rat/human chondrocyte cultures were prepared to test AT-III's effects on osteoarthritis progression and chondrocyte senescence. Potential targeted molecules of AT-III were predicted using network pharmacology and molecular docking, assessed by Western blotting and then verified with rescue experiments. AT-III treatment alleviated osteoarthritis severity (shown by OARSI grading score and micro-CT) and chondrocyte senescence (indexed by levels of SA-β-gal, P16, P53, MMP13, ROS and ratio of healthy/collapsed mitochondrial membrane potentials). Network pharmacology and molecular docking suggested that AT-III might play role through NF-κB pathway. Further experiments revealed that AT-III reduced phosphorylation of IKKα/β, IκBα and P65 in NF-κB pathway. As well as nuclear translocation of p65. Both in vivo and in vitro experiments indicated that AT-III's effects on osteoarthritis and anti-senescence were reversed by an NF-κB agonist. AT-III could alleviate osteoarthritis by inhibiting chondrocyte senescence through NF-κB pathway, which indicated that AT-III is a prospective drug for osteoarthritis treatment.

Autophagy dysregulation serves as a significant pathogenic factor in Alzheimer's disease (AD), with transcription factor EB (TFEB) acting as a pivotal transcription factor that governs the process of autophagy. Atractylenolide III (AT-III), a terpenoid compound found in medicinal Atractylodes macrocephala Koidz, is well-known for its role in antioxidant and anti-inflammatory activities. The purpose of this study is to explore the beneficial impact of AT-III on AD pathology and identify the mechanisms involved. C. elegans CL4176, SH-SY5Y APPSWE, and APP/PS1 mice were used to investigate the efficacy and possible mechanism of AT-III on the treatment of AD. AT-III reduced amyloid protein (Aβ) deposition in C. elegans CL4176 heads, prolonged the paralysis time, and reduced Aβ levels in SH-SY5Y APPSWE cells. AT-III improved the learning and memory ability of APP/PS1 mice and decreased the deposition of Aβ plaques. Transcriptomics and experimental validation showed that AT-III stimulated transcription and translation of autolysosome-associated genes. AT-III enhanced co-localization of LC3 and LAMP2 with Aβ in APP/PS1 mice. Meanwhile, AT-III increased TFEB transcriptional activity, mRNA, and protein levels in the nucleus. Furthermore, AT-III enhanced the expression of Yin Yang 1 (YY1) protein, an upstream regulator of TFEB, and led to the stimulation of autophagy and lysosome biogenesis both invivo and invitro. The observed effects were reversed upon silencing YY1. AT-III may regulate the YY1-TFEB pathway, thereby restoring autophagy flux disturbances and ameliorating AD-related pathological changes and cognitive decline. This study provides a promising lead compound for intervention in AD.

The dried rhizome of Atractylodes macrocephala Koidz. (AM) is widely utilized in traditional Chinese and Japanese Kampo medicine for its therapeutic effects including digestive improvement, spleen strengthening, and dampness reduction. Atractylenolide III (AIII), an anti-inflammatory and gastric-protective sesquiterpene of AM, serves as a critical chemical reference substance (CRS) of AM, essential for its pharmacological validation. However, its content in the decoction of traditional formula varied due to its chemical properties. This study aimed to investigate the impact of the combinations of AM and other crude drugs in traditional medicinal drug-pair theory by assessing the relationship of extracting conditions, phytochemical profiles, and the bioactivities of the decoctions prepared using various crude drug combinations. AM was decocted with the dried sclerotium of Wolfiporia cocos (WE), the dried root and rhizome of Glycyrrhiza uralensis (GU), the dried root of Panax ginseng (PG), the dried twig of Cinnamomum cassia (CC), or the dried rhizome of Zingiber officinale (ZO), respectively, with varying ratios. The extraction parameters investigated included the ratio of crude drugs-to-water, heating times, and pH of the decoction. The contents of AIII and other CRS were analyzed using HPLC. Bioactivities, including anti-inflammatory effects in RAW 264.7 cells, gastroprotective effects in gastric mucosal cells, and antioxidative effects in SH-SY5Y neuronal cells, were also evaluated. The best extracting yield of AIII from AM was achieved with the ratio of crude drugs to water 1:60 (w/v) for 30-90min of heating time at pH 5. The combinations of crude drugs significantly affected AIII turnover rates, with the highest yield obtained from formulations containing 25% AM, particularly the Lingguizhugantang (ryokeijutsukanto, LGZGT) that contains AM, WE, GU, and CC. The combination of crude drugs enhanced pharmacological activities, with LGZGT notably improving gastric mucosal protection and anti-inflammatory effects. In contrast, Shenzhetang (jinchakuto, SZT) that contains AM, WE, GU, and ZO, exhibited notable anti-inflammatory effects attributed to increased 6-gingerol content despite a reduced AIII concentration. These findings provide critical insights into optimizing AM-based formulations in traditional Chinese or Japanese Kampo medicinal theory for improving clinical efficacies and standardized phytochemical quality control.

BackgroundInflammatory reactions induced by spinal cord injury (SCI) are essential for recovery after SCI. Atractylenolide III (ATL‐III) is a natural monomeric herbal bioactive compound that is mainly derived in Atractylodes macrocephala Koidz and has anti‐inflammatory and neuroprotective effects.ObjectiveHere, we speculated that ATL‐III may ameliorate SCI by modulating microglial/macrophage polarization. In the present research, we focused on investigating the role of ATL‐III on SCI in rats and explored the potential mechanism.MethodsThe protective and anti‐inflammatory effects of ATL‐III on neuronal cells were examined in a rat SCI model and lipopolysaccharide (LPS)‐stimulated BV2 microglial line. The spinal cord lesion area, myelin integrity, and surviving neurons were assessed by specific staining. Locomotor function was evaluated by the Basso, Beattie, and Bresnahan (BBB) scale, grid walk test, and footprint test. The activation and polarization of microglia/macrophages were assessed by immunohistofluorescence and flow cytometry. The expression of corresponding inflammatory factors from M1/M2 and the activation of relevant signaling pathways were assessed by Western blotting.ResultsATL‐III effectively improved histological and functional recovery in SCI rats. Furthermore, ATL‐III promoted the transformation of M1 into M2 and attenuated the activation of microglia/macrophages, further suppressing the expression of corresponding inflammatory mediators. This effect may be partly mediated by inhibition of neuroinflammation through the NF‐κB, JNK MAPK, p38 MAPK, and Akt pathways.ConclusionThis study reveals a novel effect of ATL‐III in the regulation of microglial/macrophage polarization and provides initial evidence that ATL‐III has potential therapeutic benefits in SCI rats.

Atractylenolide Ⅲ partially alleviates tunicamycin-induced damage in porcine oocytes during in vitro maturation by reducing oxidative stress.

Integration of network pharmacology and experimental verification to reveal the active components and molecular mechanism of modified Danzhi Xiaoyao San in the treatment of depression

Involvement of mitochondrial apoptotic pathway and MAPKs/NF-κ B inflammatory pathway in the neuroprotective effect of atractylenolide III in corticosterone-induced PC12 cells

Atractylenolide-III (AIII), a sesquiterpene compound isolated from the rhizome of Atractylodes macrocephala, has been reported to have anti-inflammatory effects in the peripheral organs. However, its effects on brain inflammation remain elusive. The present study investigated the effects of AIII on the response to lipopolysaccharide (LPS) in mouse microglia and clarified the underlying mechanism. In this study, treatment of MG6 cells with AIII (100 μM) significantly decreased the mRNA expression and protein levels of toll-like receptor 4 (TLR4). In addition, pretreatment of MG6 cells and primary cultured microglia cells with AIII (100 μM) significantly decreased the mRNA expression and protein levels of tumor necrosis factor-α, interleukin-1β, interleukin-6, inducible nitric oxide synthase, and cyclooxygenase-2 induced by LPS (5 ng/mL) without cytotoxicity. Subsequently, pretreatment with AIII significantly suppressed the phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun NH2-terminal kinase (JNK) after LPS stimulation in MG6 cells. These results showed that AIII downregulated TLR4 expression, leading to suppression of the p38 MAPK and JNK pathways, which in turn inhibited the production of pro-inflammatory cytokines and enzymes in LPS-stimulated microglia. Our findings, therefore, suggest the potential for AIII as a therapeutic agent for the treatment of brain inflammation, particularly in microglia-associated inflammation.

Silica-induced apoptosis of alveolar macrophages (AMs) is an essential part of silicosis formation. Autophagy tends to present a bidirectional effect on apoptosis. Our previous study found that the blockade of autophagy degradation might aggravate the apoptosis of AMs in human silicosis. Wepresumethat targeting the autophagic pathway is regarded as a promising new strategy for silicosis fibrosis. As a main active component of the Atractylodes rhizome, Atractylenolide III (ATL-III) has been widely applied in clinical anti-inflammation. However, the effect and mechanism of ATL-III on autophagy in AMs of silicosis are unknown. In this study, we found that ATL-III might inhibit autophagy by mTOR-dependent manner, thereby improving the blockage of autophagic degradation in AMs. ATL-III alleviated the apoptosis of AMs in human silicosis. Furthermore, Rapamycin reversed the protective effect of ATL-III in AMs. These results indicate that ATL-III may be a potentially protective ingredient targeting autophagy for workers exposed to silica dust. These findings also suggest that inhibition of autophagy may be an effective way to alleviate the apoptosis of AMs in silicosis.

The performance of accelerated solvent extraction (ASE) in the analysis of atractylenolide I (AO-I) and atractylenolide III (AO-III) in Atractylodes macrocephala rhizome samples was investigated and compared with that of Soxhlet extraction (SE) and ultrasonically assisted extraction in terms of yield, extraction time, reproducibility, and solvent consumption. The results indicated that although the highest yield was achieved by SE, ASE appears to be a promising alternative to classical methods because it is faster and requires less solvent, especially when investigating large batches of plant samples. The use of ethanol at a temperature of 100 °C, performing two extraction cycles with a static duration of 10.0 min, represented the optimal ASE parameters. The extracts were analyzed by gas chromatography–triple quadrupole mass spectrometry (GC–MS/MS).