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.

This study observed the effect of Chaihu Jia Longgu Muli Decoction on myocardial fibrosis in the rat model of myocardial infarction (MI) and explored the effect of this decoction on ventricular remodeling induced by myocardial fibrosis after MI based on transforming growth factor-β1 (TGF-β1)/Smad proteins (Smads) pathway, aiming to reveal the therapeutic mechanism of this decoction. MI model rats were established by coronary artery ligation and assigned into sham, model, and low-dose, medium-dose, and high-dose (2.09, 4.19, and 8.37 g·kg^-1, respectively) Chaihu Jia Longgu Muli Decoction groups, with 10 rats in each group. Chaihu Jia Longgu Muli Decoction groups were administrated with the decoction at different doses by gavage, and the sham and model groups received an equal volume of distilled water once daily for 4 weeks. The cardiac function of rats after MI was assessed via echocardiography. Myocardial fiber deposition and collagen volume fraction (CVF) were observed by Sirius red staining and Masson staining. Ultrastructure changes of myocardial cells were observed by transmission electron microscopy. Creatine kinase isoenzymes (CK-MB), cardiac troponin T (cTnT), and soluble growth stimulation expressed gene 2 (sST2) levels were measured by ELISA. Western blot was employed to assess the protein levels of collagen Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA) associated with myocardial fibrosis, and TGF-β1, Smad3, and Smad7 associated with the TGF-β1/Smads pathway. Compared with the sham group, the model group had significantly decreased body weight, compromised cardiac function, abnormal ventricular structure, collagen fiber deposition, increased fibrosis area and CVF, seriously damaged ultrastructure of myocardium, a part of mitochondria being swollen or even ruptured, disarrangement and lysis of myofibril, and broken myotome and myofilament with emergence of space. Furthermore, the model group showed risen levels of cTnT, sST2 and CK-MB, obvious myocardial fibrosis, up-regulated protein levels of pro-fibrotic cytokines TGF-β1 and Smad3, down-regulated protein level of the inhibitory cytokine Smad7, and increased expression of myocardial fibrosis-related proteins Col Ⅰ and α-SMA. Compared with the model group, Chaihu Jia Longgu Muli Decoction groups showed increased body weight, improved cardiac function and ventricular structure, reduced collagen fiber deposition, fibrosis area, and CVF, intact ultrastructure of myocardium, normal mitochondrial structure, and well-arranged myofibril and myofilament. In addition, Chaihu Jia Longgu Muli Decoction groups presented declined cTnT, sST2, and CK-MB levels, down-regulated protein levels of TGF-β1 and Smad3, up-regulated protein level of Smad7, and reduced expression of Col Ⅰ and α-SMA. Chaihu Jia Longgu Muli Decoction may reduce excessive deposition of extracellular matrix by regulating the expression of proteins in the TGF-β1/Smads pathway, thereby alleviating myocardial fibrosis and ameliorating ventricular remodeling.

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.

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.

This study aims to investigate the mechanism by which modified Xiangsha Liujunzi Decoction (M-XS) ameliorates hepatic inflammation in hyperlipidemic mice by regulating the intestinal high-density lipoprotein 3 (HDL3) level. Intestine-specific ATP-binding cassette transporter A1 (ABCA1) knockout mice (Abcal-Villin-cre-Tg) were randomized into 3 groups (n = 8): intestine-specific knockout group (Abcal^△Vill), intestine-specific knockout mice fed a high-fat diet group (Abcal^△Vill + HFD), and intestine-specific knockout with Chinese herbal medicine intervention group (Abcal^△Vill + HFD + M-XS). Littermate control mice were randomly allocated into 3 groups (n = 8): blank group (Abcal^fl/fl), high-fat diet group (Abcal^fl/fl+HFD), and Chinese herbal medicine intervention group (Abcal^fl/fl +HFD+M-XS). Except the Abcal^fl/fl and Abcal^△Vill groups, the other groups were fed a HFD for 10 weeks. Drug administration lasted for 6 weeks at a dose of 23.66 g·kg^-1·d^-1. Serum lipid profiles (four items), hepatic histopathology (HE and oil red O staining), and ileal histopathology (HE staining) were assessed. Ileal HDL3 levels were measured by ELISA, hepatic triacylglycerol (TG) levels by the GPO-PAP method, and co-localization of CD86 and Toll-like receptor 4 (TLR4) in the liver tissue by immunofluorescence. The mRNA levels of hepatic lipopolysaccharide-binding protein (LBP), nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-κB p65), inhibitor of nuclear factor kappa-B alpha (IκBα), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) were determined by real-time PCR. The protein levels of ABCA1 and apolipoprotein A-I (apoA-I) in the ileum, as well as LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β in the liver, were assessed by Western blot. Compared with the Abcal^fl/fl group, the Abcal^fl/fl +HFD group showed increased blood lipid levels, numerous vacuoles and lipid droplets in the liver, shortened intestinal villi, elevated hepatic TG level, reduced levels of HDL3, ABCA1, and apoA-I in the ileum, co-localization of CD86 and TLR4 in the liver, and up-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β as well as protein levels of LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β in the liver (P<0.05). Compared with the Abcal^fl/fl +HFD group, the Abcal^fl/fl +HFD+M-XS group showed decreased blood lipid levels, reduced vacuoles and lipid droplets in the liver, increased intestinal villus length, decreased hepatic TG level, elevated levels of HDL3, ABCA1, and apoA-I in the ileum, weakened co-localization of CD86 and TLR4 in the liver, and down-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β and protein levels of LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β (P<0.05). Compared with the Abcal^fl/fl+HFD group, the Abcal^△Vill+HFD group showed raised blood lipid levels, increased vacuoles and lipid droplets in the liver, elevated hepatic TG level, enhanced co-localization of CD86 and TLR4 in the liver, and up-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β and protein levels of LBP, p-NF-κB, p-IκBα, TNF-α, and IL-1β (P<0.05). Compared with the Abcal^fl/fl+HFD+M-XS group, the Abcal^△Vill+HFD+M-XS group showed raised blood lipid levels, increased vacuoles and lipid droplets in the liver, elevated hepatic TG level, enhanced co-localization of CD86 and TLR4 in the liver, and up-regulated mRNA levels of LBP, NF-κB, IκBα, IL-6, TNF-α, and IL-1β and protein levels of LBP, p-NF-κB, p-IκBα, IL-6, TNF-α, and IL-1β (P<0.05). In conclusion, modified Xiangsha Liujunzi Decoction ameliorates hepatic inflammation and corrects dyslipidemia by modulating intestinal HDL3 levels.

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 investigate the effects of aged male parents on the learning ability of offspring and the intervention effect of Wuzi Yanzong Pills based on the microRNA-34a-5p (miR-34a-5p)/silent information regulator 1 (SIRT1) signaling pathway. Thirty-two SD male rats of 15 months old were randomized into aged model, model+high-dose (8 g·kg^-1) Wuzi Yanzong Pills, model+low-dose (2 g·kg^-1) Wuzi Yanzong Pills, and model+vitamin C (100 mg·kg^-1) groups (n=8). In addition, 8 SD male rats of 3 months old were selected as the control group. Rats in treatment groups were fed the diets containing different doses of Wuzi Yanzong Pills or vitamin C, and the control and model groups received a regular diet for 12 weeks. After 5 days of co-caging with 3-month-old female mice, the fertilization rate was recorded. An automated sperm analyzer was used to examine the sperm motility and count, and the testicular spermatogenesis was assessed by hematoxylin-eosin staining. The senescence cells in the testicular tissue was detected byβ-galactosidase staining, and miR-34a-5p expression was quantified via qPCR. The litter size was counted, and the body mass and body length were measured on days 1 and 30 to assess offspring development. For the offspring of 30 days old, their learning ability was examined via Morris water maze, and Nissl staining was employed to count hippocampal neurons. The miR-34a-5p expression in the hippocampal tissue of the offspring was determined by qPCR, and the protein levels of brain-derived neurotrophic factor (BDNF) and SIRT1 were determined by Western blot. Compared with the control group, the model group exhibited reductions in fertility rate, litter size, and sperm motility and count, as well as impaired testicular spermatogenesis (P<0.01). In addition, the model group showed increased senescence cells in testicular and epididymal tissue, accompanied by elevated miR-34a-5p expression in sperms. The 30-day-old offspring showed slow growth, reduced hippocampal neurons, up-regulated miR-34a-5p expression, and down-regulated protein levels of SIRT1 and BDNF in the hippocampus (P<0.01), along with impaired learning and memory performance (P<0.01). Compared with the model group, both high-dose Wuzi Yanzong Pills and vitamin C improved the fertilization rate, litter size, sperm motility, sperm count, and testicular spermatogenesis (P<0.05). The 30-day-old offspring in the two groups showed accelerated growth and development, increased hippocampal neurons, and elevated BDNF protein level in the hippocampus (P<0.05), along with enhanced learning and memory capabilities (P<0.05). Compared with the vitamin C group, the high-dose Wuzi Yanzong Pills group exhibited accelerated offspring growth (P<0.05), increases in fertilization rate and litter size (P<0.05), and improved learning and memory abilities (P<0.05). These findings indicate that Wuzi Yanzong Pills can improve testicular spermatogenesis and sperm quality in aged rats, thereby enhancing offspring's learning and memory performance. Specifically, Wuzi Yanzong Pills regulate miR-34a-5p expression to delay spermatogenic cell senescence in the testicular tissue and improve the offspring's cognitive function by miR-34a-5p mediated intergenerational transmission.

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 aims to compare the efficacy and safety of different traditional Chinese medicine injections (TCMIs) combined with conventional western medicine in the treatment of chronic obstructive pulmonary disease (COPD) with respiratory failure (RF) using a network Meta-analysis approach. A comprehensive literature search was conducted in CNKI, VIP, Wanfang, SinoMed, Web of Science, PubMed, Cochrane Library, and EMbase databases from inception to June 29, 2025. Randomized controlled trials (RCTs) were performed by employing the literature on the treatment of COPD with RF by TCMIs, and data were analyzed by Stata 17. A total of 110 RCTs involving 9 218 participants and 11 different TCMIs were included. The ranking of efficacy in the surface under the cumulative ranking curve (SUCRA) from network Meta-analysis was as follows: ① for improving clinical total response rate, Chuanxiongqin Injection + conventional western medicine, Shankezhi Injection + conventional western medicine, and Danhong Injection + conventional western medicine ranked top three; ② for reducing mortality, Chuanxiongqin Injection + conventional western medicine, Danshen Chuanxiongqin Injection + conventional western medicine, and Shenfu Injection + conventional western medicine ranked top three; ③ for shortening length of hospital stay, Danshen Chuanxiongqin Injection + conventional western medicine, Shenfu Injection + conventional western medicine, and Shenmai Injection + conventional western medicine ranked top three; ④ for reducing duration of mechanical ventilation, Shenqi Fuzheng Injection + conventional western medicine, Shenmai Injection + conventional western medicine, and Tanreqing Injection + conventional western medicine ranked top three; ⑤ for improving arterial partial pressure of oxygen (PaO₂), Chuanxiongqin Injection + conventional western medicine, Danshen Injection + conventional western medicine, and Danhong Injection + conventional western medicine ranked top three; ⑥ for reducing arterial partial pressure of carbon dioxide (PaCO₂), Shenfu Injection + conventional western medicine, Tanreqing Injection + conventional western medicine, and Danshen Chuanxiongqin Injection + conventional western medicine ranked top three; ⑦ for improving arterial oxygen saturation (SaO₂), Shenqi Fuzheng Injection + conventional western medicine, Xuebijing Injection + conventional western medicine, and Xingnaojing Injection +conventional western medicine ranked top three; ⑧ for improving forced expiratory volume in one second (FEV₁), Chuanxiongqin Injection + conventional western medicine, Shenmai Injection + conventional western medicine, and Tanreqing Injection + conventional western medicine ranked top three; ⑨ for improving forced vital capacity (FVC), Danshen Chuanxiongqin Injection + conventional western medicine, Xuebijing Injection + conventional western medicine, and Tanreqing Injection + conventional western medicine ranked top three. In terms of safety, adverse events were mostly mild and occurred at a low incidence. In conclusion, TCMIs combined with conventional western medicine may offer clinical benefits in the management of COPD with RF. However, due to limitations in the number and quality of studies for each intervention, further rigorously designed RCT should be conducted to validate these findings.