Phytomedicine最新文献

Objective: To investigate the mechanism by which Yigan Mingmu (YGMM) Decoction regulates Müller cell autophagy in diabetic retinopathy (DR).

Methods: The constituents of YGMM Decoction absorbed into the bloodstream were characterized utilizing UHPLC-HRMS analysis. Active compounds were screened via the TCMSP database to predict potential targets, while disease-associated targets were retrieved from the GeneCards and OMIM repositories. Overlapping targets were identified using Venny software, followed by network construction in Cytoscape. Subsequent analyses included PPI evaluation, GO/KEGG pathway enrichment, and molecular docking simulations. To validate the findings experimentally, a diabetic retinal edema model was established in SD rats, alongside in vitro studies using Müller cells. These experiments assessed the expression levels of specific proteins-namely Caspase-3, TXNIP, LC3-II/I, Beclin-1, p62, and mTOR-across the various treatment groups.

Results: A total of 426 active components were extracted from YGMM Decoction, and 181 active components were isolated from the serum containing YGMM Decoction. Network pharmacology analysis revealed that YGMM contains 166 active compounds targeting 240 genes, with 174 overlapping DR-related targets. Key targets included TXNIP and LC3-II. Enrichment analysis implicated oxidative stress responses and AGE-RAGE signaling. Molecular docking simulations verified a high binding affinity between the primary active ingredients (such as quercetin and kaempferol) and their respective targets. Both in vivo and in vitro studies revealed that YGMM treatment significantly downregulated the expression of Caspase-3, TXNIP, and the LC3-II/I ratio in a dose-dependent manner (p<0.05). Notably, the medium-dosage group exhibited the most optimal therapeutic efficacy.

Conclusion: UHPLC-QE-MS analysis of the main active components of YGMM and its serum-containing components demonstrated that YGMM alleviates DR by modulating autophagy via the TXNIP and LC3-II/I pathways, offering a multi-target therapeutic strategy.

Introduction: Diabetic kidney disease (DKD), a predominant driver of end-stage renal disease, intimately linked to endoplasmic reticulum stress. Echinacoside (Ech), a naturally derived phenylethanoid glycoside, exhibits diverse pharmacological activities; however, its mechanism of action in ameliorating DKD remains unclear.

Objective: To clarify the underlying molecular mechanisms of Ech's therapeutic impact on DKD.

Methods: DKD models were established using db/db mice and high glucose-induced HK-2 cells. The anti-DKD effects of Ech were systematically evaluated. Kidney proteomic sequencing was employed to screen for potential regulatory proteins, which identified Heat Shock Protein 72 (HSP72) as a core target. The IRE1/XBP1 (UPR) pathway was investigated given the established role of HSP72 as a key molecular chaperone in modulating this signaling cascade. Surface plasmon resonance (SPR), microscale thermophoresis (MST), and molecular dynamics simulations (MD) were utilized to validate interactions between Ech and HSP72. Finally, HSP72 knockdown models were generated in both in vivo and in vitro to conclusively validate Ech mechanism of action.

Results: Ech intervention ameliorated DKD in db/db mice, as evidenced by improved insulin resistance, restored glucolipid metabolic homeostasis, and attenuated kidney injury and fibrosis. Furthermore, it protected HK-2 cells against high glucose-induced cytotoxicity, reactive oxygen species (ROS)-mediated damage, and mitochondrial oxidative stress. Mechanistically, kidney proteomics revealed that Ech markedly upregulated HSP72 expression. SPR, MST, and MD results established HSP72 as a direct target of Ech, and the gene knockdown experiments demonstrated that the therapeutic efficacy of Ech against DKD was significantly abrogated in the absence of HSP72. Through direct interaction with HSP72, Ech activated the IRE1/XBP1 pathway, thereby alleviating endoplasmic reticulum stress, reducing mitochondrial oxidative stress, and suppressing apoptosis, ultimately ameliorating DKD.

Conclusion: Our study demonstrates that Ech ameliorates DKD by targeting HSP72 to activate the IRE1/XBP1 pathway, thereby mitigating mitochondrial and endoplasmic reticulum stress and reducing kidney apoptosis. These findings highlight the potential of Ech as a therapeutic agent for DKD.

Background: The effects of persistent hypoxic conditions in high-altitude regions on metabolic disorders remain poorly understood and in-depth investigation into its underlying molecular mechanisms is notably insufficient. As a metabolism-associated liver disease, the pathogenesis of non-alcoholic fatty liver disease (NAFLD) under hypoxic conditions urgently requires clarification. Previous studies have demonstrated that resveratrol (Rsv) possesses significant anti-inflammatory, antioxidant, and lipid metabolism-modulating properties, and it can act as an inhibitor of ferroptosis. Yet its precise therapeutic efficacy and mechanism in hypoxic NAFLD remain to be further explored.

Objective: This study aims to explore the potential mechanism by which Rsv improves the deterioration of NAFLD under hypoxia exposure.

Methods: Herein, we performed concurrent interventions in both in vivo and in vitro models of NAFLD, comprehensively detecting biochemical indicators (inflammatory factors, oxidative stress markers, liver function), liver histopathological changes, and nucleic acid levels to assess the effects of Rsv. By introducing ferroptosis modulators, we measured core ferroptosis parameters (mitochondrial ultrastructure, Fe²⁺, 4-HNE, LPO levels, and key protein expressions) to define disease progression patterns. By HIF-1α silencing was employed to verify its regulatory roles in ferroptosis-related factors and NAFLD pathogenesis. Co-Immunoprecipitation assays and immunofluorescence co-localization were used to explore protein interactions among ACSL4, TfR1, and HIF-1α.

Conclusion: Our results demonstrated that high-altitude hypoxia exacerbates NAFLD via inducing ferroptosis; HIF-1α upregulates the expression of key ferroptosis mediators (ACSL4, TfR1), and HIF-1α silencing attenuates ferroptosis. Rsv exerts therapeutic effects against hypoxia-related NAFLD by targeting the HIF-1α-mediated ferroptosis pathway. This study elucidates the pivotal role of HIF-1α-dependent ferroptosis in hypoxia-aggravated NAFLD, identifies the therapeutic targets and mechanisms of Rsv, and provides novel theoretical foundations and potential intervention strategies for clinical management of hypoxia-related NAFLD in high-altitude areas.