1.Proteomic Analysis of Danlou Tablet in Improving Platelet Function for Treating Coronary Heart Disease with Phlegm-stasis Intermingling Syndrome in Minipigs
Ziyan WANG ; Ying LI ; Aoao WANG ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Guoyuan ZHANG ; Lei LI ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):41-53
ObjectiveThis paper aims to observe the role of Danlou tablet in treating coronary heart disease (CHD) with phlegm-stasis intermingling syndrome in minipigs by improving platelet function and explore the potential pharmacological mechanism of Danlou tablet in regulating platelet function by using proteomics technology. MethodsThirty Bama minipigs were randomly divided into a normal control group (6 pigs) and a high-fat diet group (24 pigs). After 2 weeks of high-fat diet feeding, the high-fat diet group was randomly subdivided into a model group, an atorvastatin group (1 mg·kg-1), and Danlou tablet groups (0.6 g·kg-1 and 0.3 g·kg-1). All groups continued to receive a high-fat diet for 8 weeks after the procedure. The normal control group was given a regular diet, underwent only coronary angiography, and did not receive an interventional injury procedure. The model group and each administration group were fed a high-fat diet. Two weeks later, they underwent a coronary angiography injury procedure. After the procedure, drugs were mixed into the feed every morning for 8 consecutive weeks, with the minipigs maintained on a continuous high-fat diet during this period. Quantitative proteomics technology was further used to study platelet proteins, and differential proteins were obtained by screening. Bioinformatics analysis was performed to analyze key regulatory proteins and biological pathways involved in the therapeutic effect of Danlou tablet on CHD with phlegm-stasis intermingling syndrome. ResultsCompared with the normal control group, the model group showed a significant increase in total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) of minipigs' serum (P<0.01), a significant shortening in prothrombin time of (PT) (P<0.01), a coagulation function index, and an increase in whole blood viscosity (P<0.01) and platelet aggregation rate (P<0.01). Moreover, the platelet morphology was altered, and the contents of endothelin-1 (ET-1) and nitric oxide (NO) were significantly increased (P<0.01). Hemodynamic parameters were obviously abnormal, including significantly decreased systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), and left ventricular maximal positive dp/dt (LV+dp/dtmax) (P<0.01). Left ventricular maximal negative dp/dt (LV-dp/dtmax) was significantly increased (P<0.01). Besides, there were myocardial cell hypertrophy, obvious edematous degeneration, massive interstitial inflammatory cell infiltration, high degree of fibrosis, and coronary endothelial atherosclerosis. TC and TG levels in minipigs' serum were significantly reduced in Danlou tablet groups with 0.6 g·kg-1 and 0.3 g·kg-1 (P<0.05, P<0.01), compared with those in the model group. LDL-C was decreased in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). The whole blood viscosity under low and high shear conditions was significantly reduced in the Danlou tablet group with 0.6 g·kg-1 (P<0.05). In groups with all doses of Danlou tablet, maximum aggregation rate (MAR) and average aggregation rate (AAR) were significantly decreased (P<0.05, P<0.01), and platelets' morphological changes such as pseudopodia extension were reduced. ET-1 levels in the serum were significantly reduced. In the Danlou tablet group with 0.6 g·kg-1, NO level in the serum was reduced (P<0.05). In groups with all doses of Danlou tablet, DBP and MAP were significantly increased (P<0.05). In the Danlou tablet group with 0.6 g·kg-1, LVSP and LV+dp/dtmax were significantly increased (P<0.05, P<0.01), and LV-dp/dtmax was significantly decreased (P<0.05). In groups with all doses of Danlou tablet, edematous degeneration in myocardial tissue was milder, and coronary artery lesion degree was significantly alleviated. Compared with the normal control group, there were 94 differentially expressed proteins in the model group, including 81 up-regulated and 13 down-regulated proteins. Compared with the model group, the Danlou tablet group with 0.6 g·kg-1 showed 174 differentially expressed proteins, including 100 up-regulated and 74 down-regulated proteins. A total of 30 proteins were reversed after Danlou tablet intervention. Bioinformatics analysis revealed that its pharmacological mechanism may exert anti-platelet activation, aggregation, and adhesion effects through biological pathways such as regulation of actin cytoskeleton, platelet activation pathway, Fcγ receptor-mediated phagocytosis, as well as proteins such as growth factor receptor-bound protein 2 (GRB2), Ras-related C3 botulinum toxin substrate 2 (RAC2), RAC1, and heat shock protein 90 alpha family class A member 1 (HSP90AA1). ConclusionDanlou tablet can effectively reduce platelet activation and aggregation, exerting a good therapeutic effect on CHD with phlegm-stasis intermingling syndrome in minipigs. Its pharmacological mechanism may involve regulating biological pathways such as actin cytoskeleton and platelet activation pathway, as well as proteins like GRB2, RAC2, RAC1, and HSP90AA1, thereby exerting a pharmacological effect in anti-platelet activation, aggregation, and adhesion.
2.Xiaozheng Zhitong Paste Relieves Bone Cancer Pain in Mice by Alleviating Activation of Microglia in Spinal Cord and Damage to Neurons via Blocking PAR2/NF-κB/NLRP3 Pathway
Guangda ZHENG ; Linghan MENG ; Lu SHANG ; Juanxia REN ; Dongtao LI ; Haixiao LIU ; Lingyun WANG ; Changlin LI ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):91-100
ObjectiveTo investigate the effects and underlying mechanisms of Xiaozheng Zhitong Paste (XZP) on bone cancer pain (BCP). MethodsThirty female BALB/c mice were randomly divided into five groups: a Sham group, a BCP group, a BCP+low-dose XZP group, a BCP+high-dose XZP group, and a BCP+high-dose XZP + protease-activated receptor 2 (PAR2) agonist GB-110 group. BCP mice model was constructed by injecting Lewis lung carcinoma cells into the femoral cavity of the right leg, which was followed by being treated with XZP for 21 d. After 21 d, the mice were sacrificed. Nissl staining was used to evaluate the survival of spinal cord neurons. Immunofluorescence staining was conducted to localize ionized calcium-binding adapter molecule 1 (Iba1) and neuronal nuclear antigen (NeuN) in spinal cord tissue, thereby assessing microglial activation and neuronal survival. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), transforming growth factor-β (TGF-β), interleukin-4 (IL-4), and interleukin-10 (IL-10) in spinal cord tissue. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels associated with M1/M2 polarization of microglia. Western blot analysis was performed to examine the expression of proteins related to microglial polarization as well as those involved in the PAR2/nuclear factor kappa B (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway in the spinal cord. ResultsCompared with the Sham group, the spinal cord neurons were damaged, the number of Nissl-positive spinal cord neurons in the spinal cord tissue was significantly reduced (P<0.01), and the rate of NeuN-positive cells was significantly decreased (P<0.01). The spinal cord microglia were activated, the inflammatory level of the spinal cord tissue was enhanced, and Iba1 staining was significantly enhanced (P<0.01). The levels of IL-1β, TNF-α, IL-6, TGF-β, IL-4 and IL-10 were significantly increased (P<0.01). The mRNA expressions of IL-1β, TNF-α and inducible nitric oxide synthase (iNOS) were significantly increased (P<0.01), and the expression of PAR2, NLRP3, ASC and NF-κB p65 proteins in the spinal cord tissue of the BCP mice was significantly enhanced (P<0.01). Compared with the BCP group, high-dose XZP treatment significantly increased the number of Nissl-positive spinal cord neurons in the BCP mice (P<0.01), significantly enhanced the rate of NeuN-positive cells in the spinal cord tissue, and significantly weakened Iba1 staining (P<0.01). In addition, the levels of IL-1β, TNF-α, and IL-6 were significantly decreased, while the levels of TGF-β, IL-4, and IL-10 were significantly increased (P<0.05, P<0.01). The mRNA expression levels of IL-1β, TNF-α, and iNOS were decreased, whereas those of cluster of differentiation 206 (CD206), arginase-1 (Arg-1), and YM1/2 were significantly increased (P<0.05, P<0.01). Low-dose and high-dose XZP treatment significantly decreased the expression of PAR2, NLRP3, ASC, and NF-κB p65 proteins in the spinal cord tissue (P<0.05, P<0.01). These effects could all be significantly eliminated by the PAR2 agonist GB-110. ConclusionXZP can mitigate BCP in mice, which may be achieved through blocking the activated PAR2/NF-κB/NLRP3 pathway.
3.Discussion on Treatment of Cancer Pain with Modified Wumeiwan Based on Jueyin Syndrome
Haixiao LIU ; Linghan MENG ; Guangda ZHENG ; Dongtao LI ; Lu SHANG ; Juanxia REN ; Changlin LI ; Lingyun WANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):124-128
Pain, as one of the most common symptoms in cancer patients, seriously affects the survival quality of patients. The three-step pain relief program currently used in clinical practice cannot completely relieve pain in cancer patients and is accompanied by many problems. From the perspective of Jueyin syndrome in traditional Chinese medicine (TCM), this paper believed that the core pathogenesis of cancer pain was declined healthy Qi and cold and heat in complexity, and used Wumeiwan as the main formula with modification according to syndrome for clearing the upper, warming the lower part of the body, and harmonizing the cold and heat. It can regulate the pathological environment of deficiency, cold, stasis, toxicity, and heat, and restore the physiological function of Yang transforming Qi while Yin constituting form, so as to prevent, relieve, and even eliminate cancer pain, having achieved good clinical efficacy. It can not only help cancer patients relieve pain, but also control tumor and eliminate tumor, achieving a dual benefit of pain relief and tumor suppression. It gives full play to the characteristics and advantages of syndrome differentiation and treatment in TCM, and expands the scope of ZHANG Zhongjing's treatment for Jueyin syndrome, which provides ideas for the clinical diagnosis and treatment of cancer pain from the perspective of deficiency-excess in complexity and cold and heat in complexity.
4.Perioperative immune dynamics and clinical outcomes in patients undergoing on-pump cardiac surgery
Zhiyuan CHENG ; Xinyi LIAO ; Juan WU ; Ping YANG ; Tingting WANG ; Qinjuan WU ; Wentong MENG ; Zongcheng TANG ; Jiayi SUN ; Jia TAN ; Jing LIN ; Dan LUO ; Hao WANG ; Chaonan LIU ; Jiyue XIONG ; Liqin LING ; Jing ZHOU ; Lei DU
Chinese Journal of Blood Transfusion 2026;39(1):31-43
Objective: To characterize perioperative dynamic changes in immune-cell phenotypes and inflammatory cytokines in patients undergoing CPB (cardiopulmonary bypass) cardiac surgery, and to explore their associations with postoperative outcomes. Methods: In this prospective cohort study, 120 adult patients who underwent elective cardiac surgery under CPB at West China Hospital from May 2022 to March 2023 were enrolled. Perioperative immune-cell phenotypes and concentrations of 40 inflammation-related cytokines were measured. The primary outcomes were the sequential organ failure assessment (SOFA) score at 24 h after surgery and ΔSOFA (the peak SOFA score within 48 h after surgery minus the preoperative SOFA score). Secondary outcomes included major adverse cardiovascular events (MACE), acute kidney injury (AKI), respiratory failure, severe liver injury, and infection. Results: The mean age of enrolled patients was 57±10 years. Of these, 52% (62/120) were male and 90% (108/120) underwent valve surgery. During the rewarming to the end of CPB, neutrophil counts rapidly increased (7.39×10
/L vs preoperative 3.07×10
/L, P<0.001), with significant upregulation of CD11b (7.30×10
/L vs preoperative 3.05×10
/L, P<0.001) and CD54 (7.15×10
/L vs preoperative 2.99×10
/L, P<0.001). Lymphocyte counts increased at the end of CPB (1.75×10
/L vs preoperative 1.12×10
/L, P<0.001) but decreased significantly at 24 h after surgery (0.59×10
/L vs preoperative 1.12×10
/L, P<0.001). Plasma analysis showed that multiple pro-inflammatory cytokines increased during CPB and remained elevated up to 24 h after surgery; five chemokines and the anti-inflammatory cytokine IL-10 peaked at the end of CPB. The SOFA score increased from 1 (1, 2) preoperatively to 7 (5, 10) at 24 h after surgery, with a ΔSOFA of 6 (4, 8). Within 30 days after surgery, 48 patients (40.0%) developed AKI, 17 (14.2%) developed infection, 4 (3.3%) developed severe liver injury, 3 (2.5%) developed respiratory failure, and 3 (2.5%) experienced MACE. During the 2-year follow-up, 8 patients (6.7%) experienced MACE and 5 (4.2%) died. Conclusion: Multi-organ dysfunction is common after cardiac surgery under CPB (median ΔSOFA, 6), accompanied by perioperative activation of multiple immune-cell subsets and upregulation of pro-inflammatory, anti-inflammatory, and chemotactic mediators. This study provides data-driven evidence and research clues for further investigation of the associations between CPB-related immune perturbations and postoperative organ dysfunction and clinical outcomes.
5.Effect of dihydroquercetin on refractive development in mice with form deprivation myopia based on NLRP3 inflammasome pathway
Po ZHAO ; Meng MA ; Tengwen YANG ; Lingfei MENG ; Jing LIU ; Xiaoxiao XING ; Huiying HOU
International Eye Science 2026;26(3):391-397
AIM: To explore the effect of dihydroquercetin on visual function in mice with form deprivation myopia based on the NOD-like receptor thermoprotein domain-related protein 3(NLRP3)inflammasome pathway.METHODS: The C57BL/6 mice were randomly divided into control group and form deprivation myopia model group, and the form deprivation myopia model group was constructed by covering the right eye with a translucent eye patch. After successful modeling, the mice in the model group of form deprivation myopia were randomly divided into model group, low-, medium- and high-dose dihydroquercetin groups, and high-dose dihydroquercetin + NLRP3 agonist group. The diopter and axial length of mice in each group were detected. The kit was used to detect the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in retinal tissue. RT-qPCR was used to detect the mRNA expressions of NLRP3, apoptosis-associated spot-like protein(ASC), Caspase-1, IL-1β and IL-18 in retinal tissues. Western blot was used to detect the expression of NLRP3, ASC, cleaved Caspase-1, IL-1β and IL-18 proteins in retinal tissues. TUNEL staining was used to detect apoptosis in retinal tissue.RESULTS: Compared with the control group, the diopter of the mice in the model group decreased, and axial length increased, and the SOD decreased whereas MDA, NLRP3, ASC, Caspase-1, IL-1β, IL-18 increased, and the rate of apoptosis in retinal tissue increased(all P<0.05). Compared with the model group, the diopter of mice in the low-, medium- and high-dose dihydroquercetin groups increased, axial length shortened, the SOD increased, whereas MDA, NLRP3, ASC, Caspase-1, IL-1β, IL-18 decreased, and the rate of apoptosis in retinal tissue decreased(all P<0.05). Compared with the high-dose dihydroquercetin group, the high-dose dihydroquercetin+NLRP3 agonist group had reduced diopter, increased axial length, decreased SOD levels, elevated MDA, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 levels, as well as increased apoptosis rate in retinal tissue(all P<0.05).CONCLUSION: Dihydroquercetin can improve visual function in mice with form deprivation myopia by inhibiting pyroptosis and oxidative stress responses, which may be related to the suppression of NLRP3 inflammasome. NLRP3 agonists can partially mitigate the effects of high-dose dihydroquercetin on form deprivation myopia in mice.
6.Efficacy comparison of goniosynechialysis versus trabeculectomy combined with phacoemulsification in the treatment of glaucoma complicated with cataract
Zhentong LIU ; Nannan TIAN ; Xiangci MENG ; Yan ZHANG
International Eye Science 2026;26(3):405-409
AIM: To compare the clinical efficacy of goniosynechialysis versus trabeculectomy combined with cataract phacoemulsification in treating glaucoma complicated with cataract.METHODS:From January 2019 to January 2024, patients with glaucoma complicated with cataract diagnosed in our hospital were included as the study subjects. They were assigned into trabeculectomy group and goniosynechialysis group based on different surgical methods. The trabeculectomy group was treated with cataract phacoemulsification combined with trabeculectomy, while the goniosynechialysis group was treated with cataract phacoemulsification combined with goniosynechialysis. The improvement of visual acuity, anterior chamber depth, chamber angle width, intraocular pressure, whole en face vessel density within the optic disc head and in the peripapillary region, and complications were compared between two groups.RESULTS: A total of 120 patients(120 eyes)with glaucoma combined with cataract were included in this study, among which 60 eyes were in the trabeculectomy group and 60 eyes in the goniosynechialysis group. The age of the trabeculectomy group was 60.72±6.16 years, including 28 males and 32 females. The age of the goniosynechialysis group was 61.04±6.24 years, including 31 males and 29 females. At 3 mo after surgery, the visual acuity of patients in both groups improved significantly, and the improvement of visual acuity in the goniosynechialysis group was significantly better than that in the trabeculectomy group(all P<0.01). At 3 mo after surgery, the anterior chamber depth and chamber angle width of patients in both groups were higher than those before surgery, and with the goniosynechialysis group being higher than the trabeculectomy group(all P<0.01), while the intraocular pressure was lower than that before surgery, and with the goniosynechialysis group being lower than the trabeculectomy group(all P<0.01). Before surgery, there was no significant difference in the whole en face vessel density within the optic disc head between the two groups(P>0.05), however, the whole en face vessel density within the optic disc head in both groups on 1 d after surgery were significantly lower than before surgery(all P<0.05); and compared with 1 d after surgery, there was a significant increase at 1 wk, 1, and 3 mo after surgery(all P<0.05); the whole en face vessel density within the optic disc head in the goniosynechialysis group was significantly higher than that in the trabeculectomy group at various time points after surgery(all P<0.05). However, there was no significant difference in the whole en face vessel density in the peripapillary region between the two groups before surgery and various time points after surgery(all P>0.05). Within 3 mo after surgery, the incidence of complications in the goniosynechialysis group(8.3%)was significantly lower than that in the trabeculectomy group(21.7%; P<0.05).CONCLUSION:Goniosynechialysis combined with phacoemulsification is more effective than trabeculectomy combined with phacoemulsification in the treatment of patients with glaucoma complicated with cataract in improving the visual acuity and whole en face vessel density within the optic disc head of patients, increasing the anterior chamber depth and chamber angle width, reducing intraocular pressure and the risk of complications. It has high safety and ideal short-term efficacy.
7.The effects of galangin on the apoptosis and autophagy of gastric cancer NCI-N87 cells through regulating the AMPK/mTOR/ULK1 signaling pathway
GUO Fang ; CHEN Wei ; LIU Meng ; ZOU Yanli ; TIAN Xia
Chinese Journal of Cancer Biotherapy 2026;33(1):59-65
[摘 要] 目的:探讨高良姜素(Gal)调控AMPK/mTOR/ULK1信号通路对胃癌细胞凋亡和自噬的影响及其机制。方法:将胃癌NCI-N87细胞分为对照组、多索吗啡(DM)组、Gal低剂量(Gal-L)组、Gal高剂量(Gal-H)组、Gal-H + DM组。采用MTT法、流式细胞术、划痕愈合实验和Transwell实验分别检测各组细胞的增殖、凋亡、迁移和侵袭能力,WB法检测PCNA、C-caspase-3、免疫逃逸相关蛋白(B7H1)、EMT和AMPK/mTOR/ULK1信号通路蛋白的表达水平。建立裸鼠NCI-N87细胞移植瘤模型,观察Gal和5-FU对移植瘤的抑制效果。结果:与对照组比较,DM组NCI-N87细胞增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著升高(均P < 0.05),细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著降低(均P < 0.05);Gal-L组和Gal-H组NCI-N87细胞的增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著降低(均P < 0.05),细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著升高(均P < 0.05);DM可部分逆转Gal对NCI-N87细胞恶性生物学行为的抑制作用(P < 0.05);与对照组比较,Gal组和5-FU组裸鼠移植瘤体积和质量均显著降低,肿瘤组织细胞凋亡率显著升高(P < 0.05)。结论:Gal可促进胃癌NCI-N87细胞自噬和凋亡,抑制其增殖、迁移和侵袭,可能与激活AMPK/mTOR/ULK1信号通路有关。
8.Effect and Mechanism of Exogenous NO in Secondary Metabolism in Scutellaria baicalensis
Kai ZHAO ; Wei MA ; Weili LIU ; Zhihong LOU ; Xiangcai MENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):250-261
ObjectiveTo investigate the effects of exogenous nitric oxide (NO) on the accumulation and quality formation mechanism of flavonoids in Scutellariae Radix. MethodsFresh roots of Scutellaria baicalensis were treated with sodium nitroprusside (SNP) solutions at concentrations of 0.0, 7.5, and 20 mmol·L-1, respectively. Kits and supporting reaction systems were used to determine the following indicators of samples in each group, including (1) reactive oxygen species: changes in the content of nitric oxide (NO), superoxide anion (O
9.Research progress on cell membrane biomimetic nanoparticles for delivery of antitumor natural products
Luhua MENG ; Hong PAN ; Shuhuan LIU ; Mengmeng SHEN
China Pharmacy 2026;37(4):547-552
Natural products have shown great potential in the research and development of antitumor drugs. However, their clinical application is severely limited by inherent drawbacks such as poor water solubility, low stability, and low bioavailability. Cell membrane biomimetic nanoparticles, as a novel drug delivery system, have provided new strategies to overcome this bottleneck. This review systematically summarizes the preparation methods (e.g., membrane extrusion, ultrasonic fusion, and microfluidic electroporation) and characterization techniques (e.g., particle size, Zeta potential, and membrane surface protein detection) of cell membrane biomimetic nanoparticles, with a focus on the application of these derived from various sources in delivering antitumor natural products. Cell membrane biomimetic nanoparticles are endowed with unique biological functions, including low immunogenicity conferred by stem cell membranes, prolonged systemic circulation enabled by red blood cell membranes, and homologous targeting facilitated by tumor cell membranes. Despite these advancements, the technology still faces challenges such as difficulties in large-scale production, high costs, and limited characterization methods. Future research needs to further optimize the relevant processes to promote the clinical translation of cell membrane-biomimetic nanoparticles, thereby offering an efficient and safe novel delivery approach for antitumor therapy using natural products.
10.Effect of Epimedium brevicornu Ethanol Extract on Aging of Castrated Rats by Intervening in Mesenchymal Adipose-derived Stem Cells
Zuyu MENG ; Haiquan LIU ; Shaozi LIN ; Mei WANG ; Yiyao ZHANG ; Fang LIU ; Menghan LI ; Hongling CHEN ; Jiajia QIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):174-181
ObjectiveTo explore the mechanism by which the ethanol extract of Epimedium brevicornu (EEBM) intervenes in mesenchymal adipose-derived stem cells (ADSCs) to delay aging in castrated rats. MethodsForty-five 3-month-old SPF female SD rats were ovariectomized and randomly divided into model group, ADSCs treatment group, and ADSCs groups treated with low, medium, and high concentrations of EEBM (1, 50, 100 μg·L-1), referred to as the AE low, medium, and high concentration groups, with 9 rats in each group. After tail vein injection of 200 μL of the corresponding stem cell suspension, aging-related indicators including cyclin-dependent kinase inhibitor (p21), tumor suppressor gene (p53), interleukin-6 (IL-6), interleukin-8 (IL-8), superoxide dismutase (SOD), malondialdehyde (MDA), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), cysteine-aspartic acid protease-3 (Caspase-3), and lipofuscin were measured using enzyme-linked immunosorbent assay (ELISA) and Western blot. ResultsCompared with the model group, the IL-6 content in the AE low, medium, and high concentration groups was significantly decreased (P<0.05). Lipofuscin, MDA, and IL-8 levels in the ADSCs treatment group and AE low, medium, and high concentration groups were significantly reduced (P<0.01), while SOD content was significantly increased (P<0.05, P<0.01). Compared with the ADSCs treatment group, lipofuscin and IL-8 levels in the AE low, medium, and high concentration groups were significantly reduced (P<0.05, P<0.01). The MDA content was significantly decreased in the AE medium concentration group (P<0.01). Compared with the model group, protein levels of p21, p53, Bax, and Caspase-3 in the ADSCs treatment group and AE low, medium, and high concentration groups were significantly reduced (P<0.05, P<0.01), while the Bcl-2 protein level was significantly increased (P<0.01). Compared with the ADSCs treatment group, protein levels of p21, p53, Bax, and Caspase-3 in the AE low, medium, and high concentration groups were significantly reduced (P<0.05, P<0.01), and the Bcl-2 protein level in the AE low concentration group was significantly increased (P<0.01). ConclusionThe results of this experiment show that EEBM-treated ADSCs or ADSCs may delay aging in castrated rats by inhibiting cell apoptosis, reducing cell cycle inhibitors and pro-inflammatory factors, enhancing antioxidant capacity, and reducing oxidative reactions. Moreover, EEBM-treated ADSCs demonstrate stronger anti-aging effects than ADSCs alone. This study provides experimental evidence supporting the clinical use of EEBM to intervene in ADSCs and delay aging.

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