This study explored the mechanism by which Suanzaoren Decoction (SZRD) ameliorates hippocampal neuronal damage in APP/PS1 mice by inhibiting Bax/Bcl-2/caspase-3-mediated apoptosis through the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Differentially expressed genes (DEGs) in Alzheimer's disease (AD) were screened using the GEO database, and KEGG and GO enrichment analyses were performed using R software. Thirty 6-month-old APP/PS1 transgenic mice were randomly assigned to three groups: APP/PS1 group, low-dose SZRD group (12.96 g·kg^-1), and high-dose SZRD group (25.92 g·kg^-1), while ten age-matched male C57BL/6J mice served as the blank control group. The treatment groups received SZRD by gavage for 28 consecutive days. Following treatment, spatial learning and memory were assessed using the Morris water maze test. Neuronal numbers were evaluated by Nissl staining. Immunohistochemistry (IHC) was used to detect neuron-specific nuclear protein (NeuN) and assess amyloid-β (Aβ) deposition in hippocampal neurons. Western blot was performed to measure the expression levels of PI3K/Akt pathway proteins and apoptosis-related markers, including Bcl-2, Bax, caspase-3, and cleaved-caspase-3 in hippocampal tissues. Bioinformatics analysis identified 686 DEGs, with GO enrichment linked to synaptic function and neurotransmitter transport, and KEGG pathways including neuroactive ligand-receptor interaction and PI3K/Akt signaling. In vivo experiments showed that, compared with APP/PS1 mice, both low-and high-dose SZRD groups exhibited improved learning and memory, increased neuronal numbers, and decreased Aβ deposition. Furthermore, hippocampal levels of p-PI3K/PI3K, p-Akt/Akt, and Bcl-2 were upregulated, while Bax and cleaved-caspase-3/caspase-3 levels were downregulated. In conclusion, SZRD may improve hippocampal neuronal damage and cognitive function in APP/PS1 mice by regulating Bax/Bcl-2/caspase-3-mediated apoptosis through the PI3K/Akt signaling pathway.

This study established a hyperlipidemia model by feeding Sprague-Dawley rats a high-fat diet for 8 weeks. The rats were randomly assigned to the following groups: model group, atorvastatin calcium group (4.8 mg·kg^-1), low-, medium-, and high-dose Tanyu Tongzhi Optimization Decoction (TYTZD) groups (3.6, 7.2, and 14.4 g·kg^-1), and a normal diet control group. After 4 weeks of continuous administration, hematoxylin-eosin (HE) and oil red O staining were used to observe liver pathological changes and lipid infiltration. Automatic biochemical analyzer were performed to assess blood lipid profiles, coagulation function, and liver function. Transcriptomic and proteomic analyses were employed to identify differentially expressed genes (DEGs) and proteins (DEPs), followed by enrichment analysis. The MCODE algorithm was applied to classify DEGs and DEPs into modules, and network separation index (S_AB) was calculated to assess module separation, enabling construction of a gene-protein co-expression network for core target screening. The diagnostic accuracy of core targets was evaluated by area under the receiver operating characteristic (ROC) curve (AUC), and ELISA was used to measure core target expression. Western blot detected the expression of core pathway-related proteins in liver tissue. Results demonstrated that TYTZD significantly improved dyslipidemia, coagulation dysfunction, liver injury, hepatic pathology, and lipid infiltration in hyperlipidemic rats. Transcriptomic analysis identified 571 DEGs significantly reversed by TYTZD, mainly enriched in inflammatory signaling pathways such as Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB). Proteomic analysis identified 102 reversed DEPs, mainly involved in cholesterol metabolism pathways. Integrated analysis identified core targets including TLR4, tumor necrosis factor-α (TNF-α), integrin subunit alpha M (ITGAM), Toll-like receptor 2 (TLR2), matrix metalloproteinase 9 (MMP9), interleukin-1β (IL-1β), apolipoprotein E (APOE), and apolipoprotein C2 (APOC2), all with AUC values greater than 0.70. ELISA showed that TYTZD intervention significantly downregulated MMP9, TNF-α, IL-1β, TLR2, ITGAM, and TLR4, and upregulated APOC2 and APOE. Western blot indicated that TYTZD reduced TLR4, p-NF-κB, and IL-1β protein expression in liver tissue. In conclusion, TYTZD may exert anti-hyperlipidemic effects through regulation of core targets such as ITGAM, TLR4, and APOC2, and by modulating the TLR4/NF-κB signaling pathway to intervene in inflammatory responses and cholesterol metabolism, thereby achieving multi-target, multi-pathway therapeutic effects against hyperlipidemia.

This study aims to investigate whether the mitophagy receptor FUN14 domain-containing 1 (FUNDC1) serves as a molecular link between mitophagy and neuronal ferroptosis, and to determine whether salidroside (Sal) can inhibit neuronal ferroptosis after oxygen-glucose deprivation/reoxygenation (OGD/R) by inducing FUNDC1 expression and regulating mitophagy, thereby exerting neuroprotective effects. An in vitro model of neuronal ischemia-reperfusion injury was established with HT22 cells subjected to OGD/R. The experiment consisted of three parts: ① control, OGD/R, ferrostatin-1 (Fer-1), FUNDC1 overexpression (OV-FUNDC1), and OV-FUNDC1+Fer-1 groups; ② control, OGD/R, 3-methyladenine (3MA, an autophagy inhibitor), Sal, and Sal+3MA groups; ③control, OGD/R, Sal, FUNDC1 silencing (Si-FUNDC1), and Sal+Si-FUNDC1 groups. In the first part of the experiment, the survival rate of cells in each group was detected by the CCK-8 assay, and the protein levels of p62, microtubule-associated protein light chain 3 (LC3), acyl-CoA synthetase long-chain family 4 (ACSL4), and glutathione peroxidase 4 (GPX4) were measured by Western blot. The results showed that compared with the control group, the OGD/R group had down-regulated protein levels of p62 and GPX4 (P<0.05, P<0.01) and up-regulated protein level of ACSL4 (P<0.01); compared with the OGD/R group, the OV-FUNDC1 group showed increased protein levels of LC3 and GPX4 (P<0.05, P<0.01) and decreased protein levels of p62 and ACSL4 (P<0.05, P<0.01). In the second part of the experiment, after Sal intervention, the protein levels of FUNDC1, p62, LC3, ACSL4, and GPX4 were determined by Western blot, and changes in mitochondrial membrane potential were measured via JC-1. The results showed that compared with the OGD/R group, the Sal group had up-regulated protein levels of FUNDC1, LC3, and GPX4 (P<0.01), down-regulated protein levels of p62 and ACSL4 (P<0.01), and increased mitochondrial membrane potential (P<0.01); the 3MA group showed decreased mitochondrial membrane potential (P<0.01). In the third part of the experiment, after silencing of FUNDC1, the intracellular Fe²⁺ content was measured via a ferrous ion assay kit; reactive oxygen species (ROS) levels were measured by flow cytometry; the mitochondrial function was assessed via MitoTracker Red; adenosine triphosphate (ATP) and glutathione (GSH) levels were detected using assay kits; the expression of ACSL4 and GPX4 was detected by immunofluorescence; the protein levels of p62, LC3, ACSL4, and GPX4 were measured by Western blot. The results showed that compared with the Si-FUNDC1 group, the Sal group had decreased Fe²⁺ concentration (P<0.01), reduced ROS level (P<0.01), increased MitoTracker Red fluorescence intensity (P<0.01) and GSH content (P<0.01), weakened ACSL4 fluorescence intensity (P<0.01), enhanced GPX4 fluorescence intensity (P<0.01), up-regulated protein levels of LC3 and GPX4 (P<0.01), and down-regulated protein levels of p62 and ACSL4 (P<0.01). This study reveals that FUNDC1 may be a key protein linking mitochondrial autophagy and ferroptosis in neurons after ischemia-reperfusion injury, and Sal intervention can inhibit neuronal ferroptosis after ischemia-reperfusion by promoting the non-ubiquitinated FUNDC1-dependent mitophagy pathway.

Since its conceptual inception in 1995, Molecular Pharmacognosy has evolved over three decades from a nascent idea into a frontier interdisciplinary discipline characterized by a comprehensive theoretical system, mature methodologies, and extensive applications. This paper systematically reviews the significant milestones of this discipline in theoretical innovation, technical standardization, industrial application, and platform construction. It highlights breakthrough achievements in areas such as technical standards for the molecular identification of crude drugs, elucidation of the formation mechanisms of Dao-di herbs, establishment of conservation models for rare and endangered resources, and biosynthesis of active compounds. The paper further elucidates the evolutionary research paradigm of Molecular Pharmacognosy. Originating from phenomena observed in both the discipline and the industry, this paradigm advances through a progressive cycle of problem derivation—hypothesis presupposition—technical innovation—theoretical enrichment, driven by the dynamic interplay between theory and practice. Furthermore, the paper analyzes current challenges regarding the precision of molecular identification, the complex mechanisms of geo-authenticity, and the sustainable utilization of resources. Molecular Pharmacognosy is poised to integrate deeply with frontier technologies such as artificial intelligence, pan-omics, quantum computing, biomanufacturing, smart breeding, and gene editing. This integration will drive a revolutionary shift in the research paradigm from recognition and revelation to prediction and design. Ultimately, these achievements will mark a new phase defined by precision, systematization, and engineering, injecting fresh momentum into the modernization of TCM.

This study investigates the mechanisms and target sites of Renshen (Ginseng Radix et Rhizoma)-Danshen (Salviae Miltiorrhizae Radix et Rhizoma) (RSDS) in regulating myocardial ischemia-reperfusion injury (MIRI) via the silent information regulator 3 (SIRT3)/peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α)/voltage-dependent anion-selective channel 1 (VDAC1) signaling pathway. A rat MIRI model was established and randomly divided into control, model, fosinopril, and low-, medium-, and high-dose RSDS groups. Cardiac function, cell apoptosis, and myocardial fibrosis were assessed using cardiac ultrasound, TUNEL staining, hematoxylin-eosin (HE) staining, and Masson staining. Serum factors and the expression of key proteins and genes, such as SIRT3, PGC-1α, and VDAC1 in myocardial tissue, were analyzed using ELISA, Western blot, and RT-qPCR to investigate the underlying mechanisms through the SIRT3/PGC-1α/VDAC1 signaling pathway. The experimental results showed that, compared with the model group, RSDS significantly improved cardiac function, with the high-dose group exhibiting the most pronounced cardioprotective effect, including increased left ventricular ejection fraction (LVEF), decreased left ventricular end-systolic diameter (LVESD), and increased left ventricular fractional shortening (LVFS). TUNEL staining demonstrated that RSDS reduced myocardial cell apoptosis, and Masson staining indicated that RSDS significantly alleviated myocardial fibrosis. Serum levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), and creatine kinase-MB (CK-MB) were significantly decreased, while superoxide dismutase (SOD) levels were significantly increased, suggesting strong anti-inflammatory and antioxidant effects. Western blot and RT-qPCR analyses demonstrated that RSDS improved mitochondrial function, inhibited oxidative stress, and protected myocardial cells by regulating the SIRT3/PGC-1α/VDAC1 signaling pathway. In conclusion, RSDS significantly ameliorates cardiac dysfunction induced by MIRI through regulation of the SIRT3/PGC-1α/VDAC1 signaling pathway, reducing myocardial apoptosis, fibrosis, and inflammation.

In this study, the total alkaloids of Corydalis delicatula were isolated and purified by a combination of silica gel, aluminum oxide basic column chromatography and other chromatographic techniques. The obtained compounds were structurally identified with the help of modern spectral resolution techniques such as nuclear magnetic resonance (1D/2D NMR), high-resolution mass spectrometry (HR-ESI-MS), and single crystal X-ray diffraction. Nine alkaloids were obtained from the total alkaloids of C. delicatula, encompassing two new compounds, corlicatine (1) and N-methyl-13-oxoprotopine (2), and seven known ones: (+)-ochotensine (3), rupestrine A (4), (+)-ochotensimine (5), oxohydrastinine (6), α-magnoflorine (7), 6, 7-methylenedioxy-N-methylisoquinoline (8), and N-methyl-6, 7-dimethoxyisoquinoline (9). The inhibitory activities of the compounds against acetylcholinesterase were evaluated, with compound 4 exhibiting moderate inhibition. Molecular docking was further performed to elucidate the binding mode of this compound. The results provide a basis for elucidating the medicinal substance basis of C. delicatula.

Using bioinformatics analysis, network pharmacology, and cellular experiments, this study investigated the effects and potential mechanisms of Panax notoginseng saponins (PNS) on osteogenic induction of immortalized human aortic valve interstitial cells (hVICs). A calcific aortic valve disease (CAVD) cell model was established by treating hVICs with osteogenic induction medium (OM). Experimental groups included a control group, a model group, and PNS treatment groups (0.075, 0.05, and 0.025 mg·mL^-1). Cell viability was assessed by the CCK-8 assay. Calcification was evaluated by alkaline phosphatase (ALP) and alizarin red S staining. Western blot was performed to measure the expression of calcification-related proteins runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), and ALP. Transcriptomic datasets related to CAVD were obtained from the Gene Expression Omnibus (GEO) database, and CAVD disease targets were retrieved from GeneCards. Active components of PNS and their potential targets were collected from HERB, Batman-TCM 2.0, and ETCM databases. The intersecting genes were subjected to protein-protein interaction (PPI) network construction, Gene Ontology (GO) enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, identifying the spleen tyrosine kinase (SYK)/nuclear factor kappa-B (NF-κB) signaling pathway as a core pathway. Western blot was then used to assess the effects of PNS at the optimal concentration on SYK/NF-κB pathway-related proteins. NF-κB p65 nuclear translocation was examined by immunofluorescence co-localization, and ELISA was applied to measure tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) levels. Results showed that PNS reduced alizarin red S and ALP staining in calcification-induced hVICs and downregulated calcification-related proteins BMP2, ALP, and RUNX2 (P<0.05, P<0.01), with the most significant effect observed at 0.075 mg·mL^-1. A total of 1 416 differentially expressed genes were identified from GEO, while 5 646 CAVD-related targets were obtained from GeneCards. From HERB, Batman-TCM 2.0, and ETCM, 605 PNS targets were collected, with 130 overlapping genes identified. These were mainly involved in inflammatory responses, lipopolysaccharide-mediated signaling, transcriptional regulation, protein phosphorylation, and NF-κB pathway activation, occurring primarily in membrane rafts and the golgi apparatus, with molecular functions such as kinase activity regulation and protein binding, and enriched in lipid metabolism, atherosclerosis, and NF-κB signaling pathways. Experimental validation showed that PNS significantly decreased NF-κB p65 expression (P<0.05), significantly increased IκBα expression (P<0.01), and markedly suppressed expression of p-SYK (P<0.01), p-IκBα (P<0.01), p-NF-κB p65 (P<0.01), and p-NF-κB p50 (P<0.01). PNS also significantly reduced NF-κB p65 nuclear/cytoplasmic fluorescence intensity (P<0.01), decreased TNF-α secretion in the supernatant (P<0.01), and promoted IL-10 secretion (P<0.01). In conclusion, PNS ameliorates hVICs calcification by downregulating IκBα, NF-κB p65, and NF-κB p50 expression, inhibiting phosphorylation of SYK, IκBα, NF-κB p65, and NF-κB p50, reducing NF-κB p65 nuclear translocation, and suppressing TNF-α expression.

This study aims to explore the protective mechanism of Danqi Huayu Recipe against light-induced retinal damage (LIRD) in rats from the perspective of cell apoptosis. Sixty male SD rats were randomly allocated into six groups (n=10): normal group, model group, Ziyin Mingmu Pills group (7.8 g·kg^-1), and low-, medium-, and high-dose (3.45, 10.35, and 31.05 g·kg^-1, respectively) Danqi Huayu Recipe groups. A self-made light damage chamber was used for the modeling of LIRD in rats of other groups except normal group. Seven days after modeling, each group received corresponding agents by gavage. Normal group and model group were administrated with an equal volume of distilled water by gavage for 14 d. The retinal function was evaluated by electroretinography. Hematoxylin-eosin (HE) staining was employed to observe the retinal morphology and measure the thickness of the outer nuclear layer (ONL). The cell apoptosis was examined by terminal deoxynucleotidyl transferase dUTP nick end labeling. The ultrastructure of the retina was observed by transmission electron microscopy. The protein and mRNA levels of factors related to retinal cell apoptosis were determined by Western blot and RT-qPCR, respectively. The results showed that compared with normal group, model group demonstrated significantly decreased amplitudes of ERG a-waves and b-waves, thinned ONL, increased cell apoptosis rate, damaged retinal ultrastructure, down-regulated protein and mRNA levels of the anti-apoptotic factor (Bcl-2), and up-regulated protein and mRNA levels of pro-apoptotic factors (Caspase-3, Caspase-6, p53, and Bax). Compared with model group, Ziyin Mingmu Pills group and medium-and high-dose Danqi Huayu Recipe group recovered the amplitudes of ERG a-waves and b-waves, increased the ONL thickness, decreased the cell apoptosis rate, and improved the retinal ultrastructure. Meanwhile, they up-regulated protein and mRNA levels of Bcl-2 and down-regulated protein and mRNA levels of pro-apoptotic factors in a dose-dependent manner. In conclusion, Danqi Huayu Recipe protects rats from LIRD by regulating the expression of proteins and genes related to cell apoptosis to reduce light-induced retinal cell apoptosis.

This study aimed to investigate whether Xihuang Pills (XHP) enhance the therapeutic efficacy of temozolomide (TMZ) against glioblastoma (GBM) U251 cells by inducing mitochondria-associated ferroptosis via regulation of Nrf2/HO-1/GPX4 signaling axis. The XHP-containing serum was prepared, and the effects of various concentrations of TMZ (0-200 μmol·L^-1) and XHP (0-20%) on U251 cell viability were assessed by CCK-8 assay. The optimal synergistic dose (200 μmol·L^-1 TMZ + 10% XHP) was identified via the SynergyFinder platform for establishing a combined intervention model. Control, XHP, TMZ, and XHP + TMZ groups were designed. Cell proliferation was evaluated by EdU staining and colony formation assay, and cell migration and invasion were assessed by Transwell and wound healing assays. FerroOrange and DCFH-DA fluorescent probes were used to measure the intracellular levels of Fe²⁺ and ROS, respectively. The JC-1 fluorescent probe was used to assess mitochondrial membrane potential, while Mito-PeDPP fluorescent probe to examine mitochondrial lipid peroxidation. Mito-Tracker Green and MitoSOX^TM Red were used to examine mitochondrial morphology and oxidative stress. Transmission electron microscopy was adopted to observe the mitochondrial ultrastructure. GSH and MDA levels were measured by colorimetric assays. Western blot was employed to assess the protein levels of Nrf2, HO-1, xCT, and GPX4. The results showed that, XHP + TMZ suppressed cell viability, proliferation, migration, and invasion (P<0.01), increased Fe²⁺ accumulation, ROS generation, GSH depletion, and lipid peroxidation (P<0.001), reduced mitochondrial membrane potential, caused severe structural damage, and down-regulated the protein levels of Nrf2, HO-1, GPX4, and xCT, which indicated that the antioxidant ferroptosis-protective pathway was co-inhibited. In conclusion, XHP enhances TMZ-induced ferroptosis in GBM cells by suppressing the Nrf2/HO-1/GPX4 signaling axis. The findings provide new mechanism insights into the synergistic antitumor effect of traditional Chinese medicine in glioblastoma therapy and offer experimental evidence for the treatment of refractory brain tumors with combined strategies.

This study explored the mechanism by which the classical prescription Baihe Dihuang Decoction alleviates neuronal injury in anxious depression, based on the Src homology 2 domain-containing protein tyrosine phosphatase (SHP2) signaling pathway. Sprague-Dawley rats were randomly divided into a normal group, model group, venlafaxine group (13.5 mg·kg^-1), and Baihe Dihuang Decoction high-, medium-, and low-dose groups (16, 8, and 4 g·kg^-1, respectively). An anxious depression model was established using 28 days of chronic restraint stress (6 h/day) combined with corticosterone administration (30 mg·kg^-1, ih). From day 8 of modeling, intragastric administration of the corresponding treatments was given for 21 consecutive days. Anxiety-and depression-like behaviors were evaluated. Golgi staining was used to observe dendritic spine morphology of hippocampal neurons, and Western blot was performed to detect hippocampal phosphorylated (p)-SHP2, synapsin 1 (SYN1), and postsynaptic density protein-95 (PSD-95). In cell experiments, primary hippocampal neurons were transfected with lentiviruses to specifically overexpress or knock down SHP2. Transfection efficiency was assessed by RT-PCR and Western blot. A corticosterone-induced neuronal injury model was established, followed by intervention with Baihe Dihuang Decoction-containing serum. Cell viability was measured using the MTT assay, apoptotic morphology was observed by Hoechst 33342 staining, apoptosis rates were determined via flow cytometry, and immunofluorescence was used to detect SHP2, p-SHP2, SYN1, and PSD-95 expression. The results showed that, compared with the normal group, model rats exhibited hippocampal dendritic spines with atrophy, loss, irregular morphology, and shortening, accompanied by significantly reduced SYN1 and PSD-95 expression and markedly increased p-SHP2/SHP2 levels. After Baihe Dihuang Decoction intervention, hippocampal neuronal injury was significantly alleviated, and p-SHP2/SHP2 levels were suppressed. In vitro, neuronal SHP2 overexpression (SHP2 OE) or corticosterone modeling led to a marked decrease in cell viability, increase in apoptosis rate, and downregulation of SYN1 and PSD-95. Following treatment with Baihe Dihuang Decoction-containing serum, SHP2 overexpression was inhibited, cell viability increased, apoptosis decreased, and SYN1 and PSD-95 expression significantly increased. In addition, SHP2 knockdown (SHP2 KD) via lentiviral transfection also mimicked the protective effects of Baihe Dihuang Decoction. In conclusion, Baihe Dihuang Decoction improves hippocampal neuronal injury in anxious depression by regulating the SHP2 signaling pathway.