Usenamine A (UA) is a dibenzofuran compound isolated from Usnea longissima Arch. Previous studies have demonstrated that UA exhibited significant in vitro and in vivo anti-hepatocellular carcinoma activity, inducing both apoptosis and autophagy in human hepatocellular carcinoma cells. This study aims to further elucidate the molecular mechanism underlying its anti-hepatocellular carcinoma effect. Based on data obtained from gene chip assay, real-time quantitative PCR, and Western blot analysis, we found that UA effectively inhibited the expression of polo-like kinase 1 (PLK1) in human hepatocellular carcinoma HepG2 and SK-HEP-1 cells. Furthermore, the administration of BI 6727, a PLK1 inhibitor, significantly diminished the inhibitory effect of UA on the viability of human hepatocellular carcinoma cells. Additionally, the knockdown of PLK1 expression via RNA interference markedly inhibited the proliferation of human hepatocellular carcinoma cells, and the inhibitory effect of UA on cell viability was attenuated upon PLK1 knockdown. The knockdown of PLK1 expression significantly upregulated the apoptosis rate of human hepatocellular carcinoma cells and notably diminished the apoptosis-inducing effect of UA on these cells. Additionally, the inhibition of autophagy using the autophagy inhibitor 3-MA reduced the proliferation-inhibitory effect of UA on human hepatocellular carcinoma cells. Utilizing the PLK1 inhibitor BI 6727 or RNA interference, we further demonstrated that PLK1 negatively regulated autophagy in human hepatocellular carcinoma cells. Consequently, the inhibition of PLK1 attenuated the autophagy induction by UA on these cells. Thus, PLK1 plays a crucial role in the inhibition of human hepatocellular carcinoma cell proliferation and the induction of apoptosis by UA. Moreover, UA activates autophagy through the inhibition of PLK1, which subsequently exerts an inhibitory effect on the growth of human hepatocellular carcinoma cells.

Arjunic acid (AR), a main bioactive triterpenoid isolated from acorns, has been reported to exert pronounced anti-inflammatory activities. However, its anti-inflammatory mechanisms have not been elucidated. In this study, the model of lipopolysaccharide (LPS)-induced inflammation in RAW264.7 cells were established to investigate the anti-inflammatory activity of AR. The potent targets and signaling pathway of AR for the treatment of inflammation-related disease were predicted based on network pharmacology. Furthermore, the expression of pro-inflammatory cytokines and mediators was determined by Griess assay, enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and Western blot. The protein expression of NF-κB, MAPK, Nrf2/HO-1, PI3K/Akt/mTOR, and autophagy signaling pathways were gauged by Western blot. As the result, in the inflammatory model of LPS-induced RAW264.7 cells, AR could significantly inhibit the expression of pro-inflammatory cytokines and mediators, suppress the phosphorylation and translocation of NF-κB, and downregulate the phosphorylation of JNK/ERK signaling pathways. AR also inhibited ROS production and activated the Nrf2/HO-1 signaling pathway by degrading Keap1. Furthermore, AR activated autophagic flux by inhibiting the PI3K/Akt/mTOR signaling pathway. Collectively, AR was a potential natural product for the treatment of inflammation-related diseases.

Acute kidney injury (AKI) is a prevalent clinical syndrome characterized by a rapid deterioration in renal function. Naringin, a flavonoid abundant in the Rutaceae family, has been reported to provide protective effects against kidney injury. However, the mechanisms responsible for these effects remain inadequately elucidated. In the present study, the AKI mouse model was established in vivo through a single intraperitoneal injection of 20 mg·kg^-1 of cisplatin, and human kidney-2 (HK-2) cell injury was induced by cisplatin in vitro. Blood urea nitrogen (BUN) and creatinine (CRE) levels were measured using an ELISA kit. Hematoxylin and eosin (H&E) staining, along with periodic acid-Schiff (PAS) staining, were employed to evaluate changes in renal histopathology. Serum concentrations of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the mice were assessed using ELISA, and the levels of these cytokines were further analyzed. Morphological alterations in HK-2 cells were examined microscopically. The levels of reactive oxygen species (ROS) in HK-2 cells were quantified using an ELISA kit. The protein expression levels of p-MAPK/MAPK, p-JNK/JNK, p-ERK/ERK, p-NF-κB/NF-κB, p-IκBα/IκBα, IL-6, and TNF-α in renal tissue and HK-2 cells were evaluated through Western blot. The results showed that the serum BUN, Cre, IL-6, and TNF-α levels in the model group mice increased and the renal tissue structure was damaged when compared to the control group, whereas naringin could significantly ameliorate the above pathological changes. Moreover, naringin significantly reduced cisplatin-induced ROS production in HK-2 cells. Western blot results confirmed that naringin notably attenuated the expression of p-MAPK, p-JNK, p-ERK, p-NF-κB, p-IκBα, IL-6, and TNF-α proteins in vitro and in vivo. In conclusion, naringin may ameliorate cisplatin-induced AKI by inhibiting the activation of the MAPK/NF-κB pathway and reducing the production of ROS. All animal experiments were approved by the Institutional Animal Care and Use Committee of Hubei University of Chinese Medicine (approval No: HUCMS202204001).

This study aimed to clarify the mechanism and active components of Buyang Huanwu Decoction (BYHWD) in alleviating cerebral ischemia reperfusion injury (CIRI) by inhibiting pyroptosis. The key components and targets of BYHWD for CIRI were identified via network pharmacological analysis, followed by molecular docking performed with Autodock and Pymol software. The effects of BYHWD and its active components were validated in vivo and in vitro. A middle cerebral artery occlusion (MCAO) model was established in mouse to assess neural function alterations in mice under various conditions. Concurrently, an oxygen-glucose deprivation/reperfusion (OGD/R) model was developed utilizing mouse brain tissue astrocytes in vitro. Molecular biology experiments were used to verify the predicted key targets. We have determined that the principal components of BYHWD are baicalein and β-sitosterol. By analyzing genes associated with CIRI pathology alongside those linked to pyroptosis, 20 intersecting genes were identified. In conjunction with molecular docking binding energy assessment, TP53 and TNF emerged as pivotal core targets for subsequent validation. Molecular biology experiments confirmed that BYHWD effectively alleviates injury while reducing the expression level of P53. These findings indicate that the primary bioactive constituents of BYHWD were baicalein and β-sitosterol. In addition, BYHWD may inhibit pyroptosis via TNF and TP53 in protecting CIRI. The experiment has been approved by the Experimental Animal Welfare Ethics Committee of Zhejiang Academy of Traditional Chinese Medicine, approval number (KTSC2020037, KTSC2023030).

The study investigates the therapeutic effects and mechanisms of Buyang Huanwu Decoction (BHD) in treating ischemic stroke (IS). Using a middle cerebral artery occlusion/reperfusion (MCAO/R) rat model, we evaluated the neuroprotective effects of BHD, demonstrating significant improvements in neurological function scores, prolonged rotarod retention time, and reductions in both infarct volume and brain water content. An unsupervised clustering algorithm was employed to identify active components of BHD by clustering them with FDA-approved drugs for ischemic stroke treatment. Combined with network pharmacology analysis, the mechanisms of these active components were predicted to be associated with anti-inflammatory pathways. Further validation using a lipopolysaccharide (LPS)-induced BV-2 cell model demonstrated the anti-inflammatory efficacy of seven key active components, with their effects on anti-inflammatory activity and cell viability assessed via the Griess and MTT assays. Additionally, the content of these active components in BHD was quantified using liquid chromatography-mass spectrometry (LC-MS). In conclusion, this study elucidates the critical active components of BHD and their potential pharmacological mechanisms, providing valuable insights for the modernization of traditional Chinese medicine and its application in ischemic stroke therapy. All animal experiments were approved by the Animal and Medical Ethics Committee of Northeastern University (approval No.: NEU-EC-2023A052S).

The purpose of this study is not only to investigate the effects of Angelica sinensis polysaccharide (ASP) as a potential vaccine adjuvant on immune activation and cytokine release in RAW264.7 macrophages, but also to elucidate its underlying involved signaling mechanisms. Cell viability was evaluated by the CCK-8 assay. Flow cytometry was used to analyze the influence of ASP at five distinct concentration gradients on the expression of cluster of differentiation (CD) 80, CD86, and major histocompatibility complex Ⅱ (MHC Ⅱ) on RAW264.7 cell surfaces. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cell culture supernatant were determined by enzyme-linked immunosorbent assay (ELISA) method. Molecular techniques, including quantitative polymerase chain reaction (qPCR) were utilized to assess the mRNA expression levels of Toll-like receptor 4 (TLR4)-myeloid differentiation factor 88 (MyD88)-TNF receptor associated factor 6 (TRAF6)-nuclear factor kappa-B (NF-κB) signaling pathway. The levels of TRAF6, and the phosphorylation levels of IκB kinase (IKK) and p65 proteins were detected by Western blot. The results show that ASP at varying concentrations promote the proliferation of RAW264.7 cells without cytotoxicity. Surface molecules CD80, CD86, and MHC Ⅱ on RAW264.7 cells showed statistically significant up-regulation in response to ASP compared to the blank control (P < 0.05), with a dose-dependent effect within an optimal range. Furthermore, ASP also elevated cytokines IL-6 and TNF-α secretion levels by RAW264.7 cells compared to the normal control (P < 0.05), exhibiting a dose-response relationship within a specific concentration span. The qPCR results indicated that ASP groups at different concentrations all led to upregulation of mRNA expression levels of TLR4, MyD88, TRAF6, and NF-κB signaling pathway. The expression levels of TRAF6, p-IKK and p-p65 were increased by different concentrations of ASP. The TLR4 inhibitor TAK-242 significantly reduced the secretion of cytokines induced by APS (P < 0.05). This study highlights the immunostimulatory properties of ASP, emphasizing its potential as a vaccine adjuvant. By significantly enhancing the expression of co-stimulatory molecules and cytokines via the TLR4-MyD88-TRAF6-NF-κB signaling pathway, ASP offers a promising approach for modulating immune responses.

Twenty compounds were isolated from the twigs and leaves of the 95% ethanol extract of Miliusa tenuistipitata W. T. Wang by various chromatographic techniques including silica gel column chromatography, sephadex LH-20 and preparative HPLC. Their structures were determined by analysis of spectral data as tenuistone L (1), tenuistone K (2), sakuranetin (3), pachypodol (4), retusin (5), penduletin (6), casticine (7), 3, 7-dimetoxi-5, 3′, 4′-tri-hidroxiflavona (8), eupatin (9), (+)-miliusol (10), miliusane XIX (11), miliusane XVIII (12), miliusolide (13), 19, 20-dihydromiliusolide (14), debilisone C (15), goniothalamusin (16), 2, 10-dihydroxy-3, 9-dimethoxy-8-oxo-protoberberine (17), consanguine B (18), 6β-hydroxystigmasta-4, 22-dien-3-one (19) and 6β-hydroxystigmast-4-en-3-one (20), respectively. Among them, compounds 1 and 2 are new compounds, compounds 6, 9, 18 and 20 are isolated from Annonaceae plants for the first time and compounds 5, 7, 17 and 19 are isolated from the genus Miliusa for the first time. Human triple negative breast cancer cells (MDA-MB-231 and BT-549) were used to evaluate the antitumor activity of these isolates by MTT assay. Compounds 10 and 11 showed significant anti-proliferation activity against the test cells with half inhibition concentration (IC₅₀) of 0.98-4.85 μmol·L^-1.

In the study, we employed an untargeted metabolomic approach in conjunction with 16S rRNA high-throughput sequencing to identify potential antidepressant active effectors mediated by gut-flora among geniposides. Firstly, the rat depressed model was constructed using chronic unpredictable mild stress stimulation (CUMS) combined with orphaned model. Then the cecal contents of the rats were analyzed by untargeted metabolomics using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) in order to identify differential metabolites. The intestinal genera exhibiting significant differences in the cecal contents were identified through 16S rRNA high-throughput sequencing, and correlation analysis was subsequently conducted between the differential metabolites and the intestinal genera. The comparison of the metabolic profiles between the normal control group, the depression model group, and the geniposide treatment group revealed that 147 metabolites were down-regulated and 381 metabolites were up-regulated in the model group compared with the normal group. Furthermore, 212 metabolites were up-regulated and 288 metabolites were down-regulated in the geniposide treatment group compared with the model group. Subsequently, the combination of the screening conditions of FC (fold change) > 2, P < 0.05 and VIP (variable importance in the projection) > 1, secondary mapping and database comparison led to the identification of 55 metabolites with significant differences among the three groups. The results of the metabolic pathway enrichment analysis indicated that the identified differential metabolites were primarily involved in five metabolic pathways, namely tryptophan metabolism, arginine biosynthesis, phenylalanine biosynthesis with tyrosine and tryptophan, phenylalanine metabolism, and arginine metabolism with proline. The 16S rRNA sequencing results further indicated that Gardenia jasminoides extract may exert its antidepressant effects, thereby providing a new basis for the study of the gut-brain axis in the therapeutic strategy of depression. All animal experiments were conducted with the approval of the Biomedical Research Ethics Committee of the Naval Medical University.

Platinum-based complexes, particularly divalent platinum [platinum(Ⅱ), Pt(Ⅱ)] compounds, have become classic chemotherapy agents for the treatment of malignant tumors. However, their widespread clinical use is limited due to issues such as insufficient stability, the induction of acquired resistance and strong cytotoxicity. Although antibodies that interfere with the interaction between programmed cell death protein 1 (PD-1) and programmed cell death ligand 1 (PD-L1), in combination with platinum-based compounds, have shown significant clinical progress in cancer treatment, their high efficacy is often accompanied by substantial toxicity and immune-related side effects, which limit their long-term use. In contrast, platinum(Ⅳ) [Pt(Ⅳ)] complexes, with their unique octahedral geometry, have demonstrated promising anticancer potential. By modifying the axial ligands, Pt(Ⅳ)-based complexes not only show higher inertness and improved tumor selectivity, but also enable the targeted release of active ligands in the tumor microenvironment. The mechanism allows Pt(Ⅳ)-based complexes to overcome drug resistance, reduce toxicity and enhance immune system activation, making them a research hotspot in the current cancer field. More importantly, Pt(Ⅳ)-based complexes can exert anticancer effects through multiple pathways including causing DNA damage to trigger apoptosis, autophagy and ferroptosis in tumor cells. This multifaceted action mechanism not only enhances antitumor efficacy but also significantly reduces side effects associated with traditional platinum-based compounds. This review summarizes whole anticancer mechanisms of platinum-based complexes, particularly Pt(Ⅳ) complexes in chemo-immunetherapy combination therapies, discussing their potential in cancer treatment and providing theoretical support for the development of efficient, low-toxicity and highly selective Pt(Ⅳ)-based complexes.

Transient receptor potential vanilloid (TRPV), an important class of non-selective cation channels, is closely associated with a variety of physiological and pathological processes. Pungent flavour is one of the important flavours in the doctrine of five flavours in traditional Chinese medicine, which has the functions of relieving the epidermis, moving the Qi, activating the blood and so on, which has a variety of main treatments. Modern research has shown that the active ingredients in numerous pungent Chinese medicine have the ability to modulate TRPV channels. The purpose of paper is to explore the biological characteristics of TRPV channels, the modern research process of pungent Chinese medicine and its mechanism in inflammation, pain and other pathological processes. This paper discusses the research progress of the pharmacological action of pungent Chinese medicine based on TRPV channel, in order to reveal the scientific connotation of spicy traditional Chinese medicine, and also provide new research ideas and models for the modern research of Chinese medicine.