1.Strategies for Building an Artificial Intelligence-Empowered Trusted Federated Evidence-Based Analysis Platform for Spleen-Stomach Diseases in Traditional Chinese Medicine
Bin WANG ; Huiying ZHUANG ; Zhitao MAN ; Lifeng REN ; Chang HE ; Chen WU ; Xulei HU ; Xiaoxiao WEN ; Chenggong XIE ; Xudong TANG
Journal of Traditional Chinese Medicine 2026;67(1):95-102
This paper outlines the development of artificial intelligence (AI) and its applications in traditional Chinese medicine (TCM) research, and elucidates the roles and advantages of large language models, knowledge graphs, and natural language processing in advancing syndrome identification, prescription generation, and mechanism exploration. Using spleen-stomach diseases as an example, it demonstrates the empowering effects of AI in classical literature mining, precise clinical syndrome differentiation, efficacy and safety prediction, and intelligent education, highlighting an upgraded research paradigm that evolves from data-driven and knowledge-driven approaches to intelligence-driven models. To address challenges related to privacy protection and regulatory compliance in cross-institutional data collaboration, a "trusted federated evidence-based analysis platform for TCM spleen-stomach diseases" is proposed, integrating blockchain-based smart contracts, federated learning, and secure multi-party computation. The deep integration of AI with privacy-preserving computing is reshaping research and clinical practice in TCM spleen-stomach diseases, providing feasible pathways and a technical framework for building a high-quality, trustworthy TCM big-data ecosystem and achieving precision syndrome differentiation.
2.Textual Research of Key Information of Classic Formula Xieqingwan Based on Ancient and Modern Literature
Yujie CHANG ; Lyuyuan LIANG ; Jialei CAO ; Xinghang LYU ; Wenxi WEI ; Xiaofang WANG ; Huizhen ZHANG ; Sai REN ; Mengqi WANG ; Bingqi WEI ; Bingxiang MA
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):226-234
ObjectiveThis paper aims to systematically collect and organize ancient and modern clauses and studies containing Xieqingwan, excavate and analyze the key information of Xieqingwan, and provide a reference for facilitating the development of the classic formula Xieqingwan. MethodsThe composition, dosage, decocting methods, usage, and other key information of Xieqingwan in ancient traditional Chinese medicine books were collected and analyzed by means of literature research and metrological methods. The modern clinical application of Xieqingwan was summarized. ResultsA total of 42 pieces of effective data involving 32 ancient traditional Chinese medicine books were collected. Xieqingwan was first recorded in Xiaoer Yaozheng Zhijue. The drug origin of this formula is basically clear in the ancient traditional Chinese medicine books. The modern drug usage and decocting method were as follows: Angelicae Sinensis Radix, Gentianae Radix et Rhizoma, Chuanxiong Rhizoma, Gardenia seeds, Radix et Rhizoma Rhei, Notopterygii Rhizoma et Radix, and Saposhnikoviae Radix were grounded to fine powder, decocted with honey, and finally formed into pills with the size of a chicken head (1.5 g). It was suggested that half a pill or one pill were taken for one dose with warm Lophatheri decoction and sugar. The indications and clinical application had developed from the recordings in Xiaoer Yaozheng Zhijue and evolved from pediatrics to ophthalmic otolaryngology, neurology, dermatology, digestion, and respiratory diseases. The main pathogenesis of these diseases is heat in the liver meridian and is treated. The effect of Xieqingwan is "clearing away heat and toxicity, removing fire and relaxing the bowels, and dispersing swelling and relieving pain". It is recommended to use the corresponding preparation methods in the 2020 Edition of Pharmacopoeia of the People's Republic of China. Modern clinical studies are centered around the clinical application of Xieqingwan, which is often modified and used in treating Tourette syndrome, herpes, febrile convulsion, sleepwalking, and insomnia. ConclusionThis paper conducts a thorough textual research of the key information of Xieqingwan, induces its historic evolution, and confirms its key information, so as to provide a reference for the future development of Xieqingwan.
3.Analysis and Discussion of Traditional Chinese Medicine and Compounds to Improve Diabetic Cardiomyopathy by Regulating Cardiomyocyte Pyroptosis
Ying ZHANG ; Chengzhi XIE ; Chang FU ; Jianxun REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):260-267
Diabetic cardiomyopathy (DCM) is a myocardium-specific microvascular disease caused by diabetes mellitus that impairs the structure and function of the heart. It is the major cause of morbidity and mortality in diabetic individuals. Traditional Chinese medicine (TCM) has extensive clinical experience and precise efficacy in treating DCM, and its multi-target, multi-pathway, multi-component, and low side effect approach can slow the progression of DCM and improve the symptoms while effectively dealing with the complexity and long-term nature of its pathological process. Many recent studies have demonstrated that pyroptosis accompanied by inflammatory response is one of the main types of myocardial injury in DCM, which promotes the development of DCM and is closely related to pathological changes such as oxidative stress, myocardial fibrosis, myocardial hypertrophy, and decreased cardiac function in the course of DCM. These findings also provide a theoretical foundation for future research into potential therapy techniques and intervention mechanisms for DCM. By searching and analyzing relevant literature from several databases, including CNKI, PubMed, Web of Science, Excerpt Medica, Science Direct, and Springer, this study aimed to comprehensively analyze the characteristics of the effects of TCM and compounds in intervening in cardiomyocyte pyroptosis in DCM in recent years and explore the potential mechanisms. It also reveals the potential of effective components of TCM and compounds in preventing and controlling DCM from the standpoint of cardiomyocyte pyroptosis and provides a new way of thinking and more experimental evidence for the clinical application of TCM in treating DCM.
4.PDGF-C: an Emerging Target in The Treatment of Organ Fibrosis
Chao YANG ; Zi-Yi SONG ; Chang-Xin WANG ; Yuan-Yuan KUANG ; Yi-Jing CHENG ; Ke-Xin REN ; Xue LI ; Yan LIN
Progress in Biochemistry and Biophysics 2025;52(5):1059-1069
Fibrosis, the pathological scarring of vital organs, is a severe and often irreversible condition that leads to progressive organ dysfunction. It is particularly pronounced in organs like the liver, kidneys, lungs, and heart. Despite its clinical significance, the full understanding of its etiology and complex pathogenesis remains incomplete, posing substantial challenges to diagnosing, treating, and preventing the progression of fibrosis. Among the various molecular players involved, platelet-derived growth factor-C (PDGF-C) has emerged as a crucial factor in fibrotic diseases, contributing to the pathological transformation of tissues in several key organs. PDGF-C is a member of the PDGFs family of growth factors and is synthesized and secreted by various cell types, including fibroblasts, smooth muscle cells, and endothelial cells. It acts through both autocrine and paracrine mechanisms, exerting its biological effects by binding to and activating the PDGF receptors (PDGFRs), specifically PDGFRα and PDGFRβ. This binding triggers multiple intracellular signaling pathways, such as JAK/STAT, PI3K/AKT and Ras-MAPK pathways. which are integral to the regulation of cell proliferation, survival, migration, and fibrosis. Notably, PDGF-C has been shown to promote the proliferation and migration of fibroblasts, key effector cells in the fibrotic process, thus accelerating the accumulation of extracellular matrix components and the formation of fibrotic tissue. Numerous studies have documented an upregulation of PDGF-C expression in various fibrotic diseases, suggesting its significant role in the initiation and progression of fibrosis. For instance, in liver fibrosis, PDGF-C stimulates hepatic stellate cell activation, contributing to the excessive deposition of collagen and other extracellular matrix proteins. Similarly, in pulmonary fibrosis, PDGF-C enhances the migration of fibroblasts into the damaged areas of lungs, thereby worsening the pathological process. Such findings highlight the pivotal role of PDGF-C in fibrotic diseases and underscore its potential as a therapeutic target for these conditions. Given its central role in the pathogenesis of fibrosis, PDGF-C has become an attractive target for therapeutic intervention. Several studies have focused on developing inhibitors that block the PDGF-C/PDGFR signaling pathway. These inhibitors aim to reduce fibroblast activation, prevent the excessive accumulation of extracellular matrix components, and halt the progression of fibrosis. Preclinical studies have demonstrated the efficacy of such inhibitors in animal models of liver, kidney, and lung fibrosis, with promising results in reducing fibrotic lesions and improving organ function. Furthermore, several clinical inhibitors, such as Olaratumab and Seralutinib, are ongoing to assess the safety and efficacy of these inhibitors in human patients, offering hope for novel therapeutic options in the treatment of fibrotic diseases. In conclusion, PDGF-C plays a critical role in the development and progression of fibrosis in vital organs. Its ability to regulate fibroblast activity and influence key signaling pathways makes it a promising target for therapeutic strategies aiming at combating fibrosis. Ongoing research into the regulation of PDGF-C expression and the development of PDGF-C/PDGFR inhibitors holds the potential to offer new insights and approaches for the diagnosis, treatment, and prevention of fibrotic diseases. Ultimately, these efforts may lead to the development of more effective and targeted therapies that can mitigate the impact of fibrosis and improve patient outcomes.
5.Interactions between Xuefu Zhuyu Decoction and atorvastatin based on human intestinal cell models and in vivo pharmacokinetics in rats.
Xiang LI ; Huan YI ; Chang-Ying REN ; Hao-Hao GUO ; Hong-Tian YANG ; Ying ZHANG
China Journal of Chinese Materia Medica 2025;50(11):3159-3167
The study aims to explore the herb-drug interaction between Xuefu Zhuyu Decoction(XFZY) and atorvastatin(AT). Reverse transcription polymerase chain reaction(RT-PCR) was used to analyze the transcription levels of proteins related to drug metabolism and transport in LS174T cells, detect the intracellular drug uptake under various substrate concentrations and incubation time, and optimize the model reaction conditions of transporter multidrug resistance protein 1(MDR1)-specific probe Rhodamine 123 and AT to establish a cell model for investigating the human intestinal drug interaction. The cell counting kit-8(CCK-8) method was adopted to evaluate the cytotoxicity of XFZY on LS174T cells. After a single and continuous 48 h culture with XFZY, AT or Rhodamine 123 was added for co-incubation. The effect and mechanism of XFZY on human intestinal absorption of AT were analyzed by measuring the intracellular drug concentrations and transcription levels of related transporters and metabolic enzymes. The results of in vitro experiments show that a single co-culture with a high concentration of XFZY significantly increases the intracellular concentrations of Rhodamine 123 and AT. A high concentration of XFZY co-culture for 48 h increases the AT uptake level, significantly induces the CYP3A4 and UGT1A1 gene expression levels, and inhibits the OATP2B1 gene expression level. To compare with the evaluation results of the in vitro human cell model, the pharmacokinetic experiment of XFZY combined with AT was carried out in rats. Sprague-Dawley(SD) rats were randomly divided into a blank control group and an XFZY group. After 14 days of continuous intragastric administration, AT was given in combination. The liquid chromatography-mass spectrometry(LC-MS)/MS method was used to detect the concentrations of AT and metabolites 2-hydroxyatorvastatin acid(2-HAT), 4-hydroxyatorvastatin acid(4-HAT), atorvastatin lactone(ATL), 2-hydroxyatorvastatin lactone(2-HATL), and 4-hydroxyatorvastatin lactone(4-HATL) in plasma samples, and the pharmacokinetic parameters were calculated. Pharmacokinetic analysis in rats shows that continuous administration of XFZY does not significantly change the pharmacokinetic characteristics of AT in rats, but the AUC_(0-6 h) values of AT and metabolites 2-HAT, 4-HAT, and 2-HATL increase by 21.37%, 14.94%, 12.42%, and 6.68%, respectively. The metabolic rate of the main metabolites shows a downward trend. The study indicates that administration combined with XFZY can significantly increase the uptake level of AT in human intestinal cells and increase the exposure level of AT and main metabolites in rats to varying degrees. The mechanism may be mainly due to the inhibition of intestinal MDR1 transport activity.
Animals
;
Drugs, Chinese Herbal/administration & dosage*
;
Atorvastatin/administration & dosage*
;
Humans
;
Rats
;
Rats, Sprague-Dawley
;
Male
;
Intestines/cytology*
;
Intestinal Mucosa/metabolism*
;
Herb-Drug Interactions
;
Cytochrome P-450 CYP3A/metabolism*
;
Intestinal Absorption/drug effects*
6.Study on the in vivo intestinal absorption and tissue distribution of silybin nanocrystals prepared by two methods
Mengyan WANG ; Ying SUN ; Sirui HUANG ; Yabo REN ; Jinhua CHANG ; Xigang LIU
China Pharmacy 2025;36(11):1335-1339
OBJECTIVE To investigate the absorption characteristics and tissue distribution of silybin (Sy) nanocrystals prepared by two methods in different intestinal segments of rats. METHODS Sy nanocrystals (i.e. Sy-NS-G and Sy-NS-F) with comparable particle sizes were prepared using high-pressure homogenization and anti-solvent precipitation methods, respectively. Rats were randomly divided into three groups: Sy raw drug group, Sy-NS-G group, and Sy-NS-F group. Each group was further divided into three subgroups with low, medium, and high (60, 120, 180 μg/mL) mass concentrations (calculated based on Sy), with 3 rats in each subgroup. The absorption rate constant (Ka) and apparent absorption coefficient (Papp) of Sy raw drug, Sy-NS-G and Sy-NS-F in different intestinal segments were investigated by using the in vivo one-way intestinal perfusion experiment. Additionally, the rats were divided into three groups: Sy raw drug group, Sy-NS-G group, and Sy-NS-F group, with 20 rats in each group. Rats in each group were administered a single intragastric dose of 50 mg/kg (calculated based on Sy). They were sacrificed at 0.3, 1, 4, 10, and 24 hours post-administration respectively, to investigate the tissue distribution of Sy raw drug, Sy- NS-G, and Sy-NS-F in the heart, liver, spleen, lungs, kidneys, brain and intestines. RESULTS In duodenum and jejunum, the Ka and Papp of the nanocrystals prepared by the two methods remained unchanged with the increase of Sy concentration, and there was no significant difference (P>0.05); the absorption of Sy-NS-F in the duodenum was greater than that of Sy-NS-G; the absorption sites of Sy-NS-G and Sy raw drug were mainly in the ileum, while those of Sy-NS-F were mainly in the duodenum and ileum. The concentrations of Sy-NS-G and Sy-NS-F in different tissues of rats were different; Sy-NS-G peaked in most tissues at 1 h, and the distribution concentration was as follows: intestine>spleen>heart>lungs>liver≈brain>kidneys. Sy-NS-F reached its peak at 1 h, and the distribution concentration was in the order of intestine>spleen>kidney>lung>heart≈liver>brain. CONCLUSIONS The absorption mode of Sy nanocrystals in the duodenum and ileum is mainly passive diffusion. In the duodenum, the absorption of Sy-NS-F is greater than that of Sy-NS-G; there are significant differences in the tissue distribution of Sy-NS-G and Sy-NS-F in rats.
7.Evaluation progress of the application of staplers in thoracoscopic lung surgery
Shenghui LI ; Yijiu REN ; Hang SU ; Minglei YANG ; Guofang ZHAO ; Yongxiang SONG ; Xuefei HU ; Deping ZHAO ; Qi XUE ; Chang CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(05):709-713
Compared to traditional suturing, lung stapling using automatic staplers offers advantages such as smaller trauma, faster wound healing, ease of operation, and lower complication rates, making it widely used in clinical practice. However, there are significant differences in bronchial tissue thickness at different anatomical locations, and the market is flooded with various types of staplers. Currently, there is a lack of recommended stapling schemes for bronchial staplers at different anatomical locations. This article reviews the development and application of automatic staplers and summarizes some types of staplers that are currently used in clinical practice, with the aim of promoting the formation of individualized stapler selection protocols for minimally invasive thoracic surgery based on the Chinese population.
8.Effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture on reproductive function in mice with asthenozoospermia based on mitochondrial apoptosis.
Jianheng HAO ; Boya CHANG ; Jia REN ; Zhen GAO ; Yanlin ZHANG ; Haijun WANG ; Laixi JI
Chinese Acupuncture & Moxibustion 2025;45(1):71-81
OBJECTIVE:
To observe the effects of the "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture on key regulatory factors during mitochondrial apoptosis of testicular tissue in asthenozoospermia mice, and explore the potential mechanism of the protective effect of acupuncture on reproductive function.
METHODS:
Thirty C57BL/6 male mice were randomly divided into a blank group, a model group and an acupuncture group, 10 mice in each group. In the model and the acupuncture groups, the intraperitoneal injection of cyclophosphamide (30 mg•kg-1•d-1) was delivered for 7 days to prepare the asthenozoospermia model. After the success of modeling, the modeled mice in the acupuncture group were intervened with "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture, once daily and the needles were retained for 20 min. The duration of the intervention was 2 weeks. The general condition of each mouse was observed, and the body mass was recorded before modeling, after modeling and after intervention completion. After intervention, the testicular mass was recorded and the weight coefficient was calculated, and the mouse sperm quality was examined; the serum contents of testosterone (T), follicle stimulating hormone (FSH) and luteinizing hormone (LH) were detected using ELISA, the morphology of testicular tissue was observed using HE, the mitochondrial ultra-microstructure of testicular tissue was observed under transmission electrone microscopy, the mitochondrial membrane potential level of testicular tissue was detected using JC-1 staining, the positive rate of apoptosis cell of testicular tissue was observed using TUNEL; and the mRNA and protein expression of b-cell lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cytochrome c (Cyt C), apoptotic protease-activating factor1 (Apaf-1), Caspase-9 and Caspase-3 of testicular tissue was detected using real-time quantitative fluorescence PCR and Western blot methods separately; and the positive expression of Cleaved Caspase-3 of the testicular tissue was detected using immunohistochemistry.
RESULTS:
Compared with the blank group, the mice were in listless spirits, had shaggy hairs, the reduced appetite and movement, and weight loss in the model group (P<0.01); the testicular mass and the weight coefficient decreased (P<0.01); the total number of sperms, sperm motility, and sperm viability were declined (P<0.01); while the levels of serum T, FSH, and LH were dropped (P<0.01). The morphology of seminiferous tubules in testicular tissue was abnormal, the number of spermatogenic cells and the number of mitochondria decreased, the inner mitochondrial crest was fractured and lost, and vacuoles appeared. The level of mitochondrial membrane potential was reduced (P<0.01); and the positive rate of apoptosis cell in testicular tissue increased (P<0.01). The mRNA and protein expression of Bax, Cyt C, Apaf-1, Caspase-9 and Caspase-3 was elevated (P<0.01, P<0.05), the mRNA and protein expression of Bcl-2 was dropped (P<0.01), and the average absorbance value of Cleaved Caspase-3 increased (P<0.01). When compared with the model group, in the acupuncture group, the general condition of mice was improved, the testicular mass and the weight coefficient elevated (P<0.01); the total number of sperms, sperm motility, and sperm viability increased (P<0.01); while the levels of serum T, FSH, and LH rose (P<0.01). The pathological morphology of testicular tissue and the inner mitochondrial ultra-microstructure were ameliorated, the level of mitochondrial membrane potential was elevated (P<0.01); the positive rate of apoptosis cell was reduced (P<0.01). The mRNA and protein expression of Bax, Cyt C, Apaf-1, Caspase-9 and Caspase-3 was dropped (P<0.01, P<0.05), the mRNA and protein expression of Bcl-2 elevated (P<0.05), and the average absorbance value of Cleaved Caspase-3 declined (P<0.01).
CONCLUSION
"Zhibian" (BL54)-toward- "Shuidao" (ST28) acupuncture may ameliorate mouse reproductive function by inhibiting mitochondrial apoptosis pathway, alleviating testicular tissue damage in the asthenospermia mice induced by cyclophosphamide.
Animals
;
Male
;
Mice
;
Apoptosis
;
Acupuncture Therapy
;
Mitochondria/metabolism*
;
Asthenozoospermia/genetics*
;
Humans
;
Testis/metabolism*
;
Mice, Inbred C57BL
;
Spermatozoa/metabolism*
;
Acupuncture Points
;
Sperm Motility
;
Testosterone/blood*
;
Proto-Oncogene Proteins c-bcl-2/genetics*
;
Caspase 3/genetics*
;
Follicle Stimulating Hormone/blood*
;
Reproduction
;
Cytochromes c/genetics*
;
bcl-2-Associated X Protein/genetics*
;
Apoptotic Protease-Activating Factor 1/genetics*
9.Analgesic effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique of acupuncture on primary dysmenorrhea based on NOD1/RIP2/NF-κB signaling pathway in the rats.
Xu JIN ; Yanlin ZHANG ; Boya CHANG ; Jia REN ; Jianheng HAO ; Yuxia CAO ; Haijun WANG ; Laixi JI
Chinese Acupuncture & Moxibustion 2025;45(2):209-216
OBJECTIVE:
To observe the effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique on the relative protein expression of the signaling pathway of nucleotide-binding oligomerization domain-containing protein 1 (NOD1)/ receptor-interacting protein 2 (RIP2)/nuclear factor kappa-B (NF-κB) and the expression of proinflammatory cytokines in the rats with primary dysmenorrhea (PD), so as to explore the underlying mechanism of this acupuncture technique for pain alleviation in PD.
METHODS:
Thirty female SD rats of SPF grade with normal estrous cycle were randomized into a blank group, a model group and an acupuncture group, 10 rats in each one. Using the intraperitoneal injection with estradiol benzoate combined with oxytocin, PD model was prepared in the model group and the acupuncture group. In the acupuncture group, during model preparation, the intervention with "Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique was delivered simultaneously, 20 min each time, once daily for consecutive 10 days. On day 11, within 30 min after the intraperitoneal injection with oxytocin, the writhing reaction (latency, frequency and score) was recorded; the morphology of uterine tissue was observed with HE staining, the contents of prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α), interleukin (IL)-1β, IL-18, cyclooxygenase-2 (COX-2), and tumor necrosis factor-α(TNF-α) in the serum were detected using ELISA method; the relative protein expression of NOD1, RIP2, NF-κB p65, phosphorylation-NF-κB p65 (p-NF-κB p65) was detected in the uterine tissue using Western blot method; and the mRNA expression of NOD1, RIP2 and NF-κB p65 was detected with the quantitative real-time PCR employed.
RESULTS:
Compared with the blank group, in the model group, the writhing latency was prolonged (P<0.01), the writhing frequency and score increased (P<0.01) in the rats; the endometrial epithelial cells showed massive degeneration and necrosis, with severe endometrial edema and widespread shedding, combined with neutrophil infiltration; the serum PGE2 content was dropped (P<0.01), while those of PGF2α, IL-1β, IL-18, COX-2, and TNF-α elevated (P<0.01); the protein expression of NOD1, RIP2, NF-κB p65 and p-NF-κB p65, and the mRNA expression of NOD1, RIP2 and NF-κB p65 in uterine tissue increased (P<0.01). In comparison with the model group, in the acupuncture group, the writhing latency was prolonged (P<0.01), the writhing frequency and score were reduced (P<0.01) in the rats; there was less degeneration and necrosis of endometrial epithelial cells, with mild endometrial edema and very little neutrophil infiltration; the serum PGE2 content increased (P<0.01), while those of PGF2α, IL-1β, IL-18, COX-2, and TNF-α decreased (P<0.01); the protein expression of NOD1, RIP2, NF-κB p65 and p-NF-κB p65 and the mRNA expression of NOD1, RIP2 and NF-κB p65 in uterine tissue were dropped (P<0.05, P<0.01).
CONCLUSION
"Zhibian" (BL54)-toward-"Shuidao" (ST28) needling technique can alleviate the pain symptom of PD rats, and its action mechanism may be related to inhibiting the active expression of NOD1/RIP2/NF-κB signaling pathway in the uterine tissue, thereby reducing the inflammatory response.
Animals
;
Female
;
Rats, Sprague-Dawley
;
Rats
;
Signal Transduction
;
Dysmenorrhea/metabolism*
;
NF-kappa B/metabolism*
;
Acupuncture Points
;
Humans
;
Acupuncture Analgesia
;
Nod1 Signaling Adaptor Protein/metabolism*
;
Receptor-Interacting Protein Serine-Threonine Kinase 2/metabolism*
;
Acupuncture Therapy
10.Mechanism research of "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture technique for improving reproductive function in mice with asthenospermia based on the ferroptosis pathway.
Jianheng HAO ; Boya CHANG ; Qingkai JIN ; Jia REN ; Haijun WANG ; Laixi JI
Chinese Acupuncture & Moxibustion 2025;45(3):351-360
OBJECTIVE:
To investigate the underlying mechanism of "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture technique for improving reproductive function in mice with asthenospermia by regulating ferroptosis pathway.
METHODS:
Sixty male C57BL/6 mice were randomly divided into a blank group, a model group, an acupuncture group and a Fer-1 group, 15 mice in each one. Except the blank group, the intraperitoneal injection with cyclophosphamide (50·kg-1·d-1) was administered to establish the asthenospermia model in the mice of the rest 3 groups for 5 consecutive days. In the acupuncture group, "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture technique was operated in the mice, for 20 min each time; and in the Fer-1 group, Fer-1 solution (1 mg/kg) was injected intraperitoneally. The interventions of these two groups were delivered once daily and for 2 consecutive weeks. The testicular wet weight was measured and the testicular coefficient was calculated. Using sperm quality detection system, the sperm quality was detected. With ELISA used, the contents of testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the serum were detected. With HE staining, testicular and epididymal morphology was observed. Immunofluorescence was used to detect the expression of reactive oxygen species (ROS) in the testes. Biochemical assay was conducted to determine the contents of malondialdehyde (MDA), reduced glutathione (GSH), and total iron ion (TFe) in the testicular tissue. Transmission electron microscopy was used to examine mitochondrial structure of the testis, while JC-1 staining was used to assess mitochondrial membrane potential in the testicular tissue. Fluorescence quantitative PCR and Western blot analyses were employed to measure the mRNA and protein expression of solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), and acyl-CoA synthetase long-chain family member 4 (ACSL4) in the testicular tissue.
RESULTS:
Compared with the blank group, in the model group, the testicular wet weight and testicular coefficient decreased (P<0.01); the sperm concentration and sperm motility reduced (P<0.01), and the contents of T, FSH, and LH decreased in the serum (P<0.01); and the seminiferous tubules in the testis showed loose structure and deformed lumen, sperm cells were disorganized and the sperm numbers reduced; the tubular walls became thinner, and sperm numbers in the lumen less; the expression of ROS in testicular tissue, as well as the contents of MDA and TFe increased (P<0.01), and the content of GSH decreased (P<0.01); and the numbers of mitochondria reduced, the structure of cristae was serious damaged, and mitochondrial membrane potential level declined (P<0.01); the mRNA and protein expression of SLC7A11, GPX4, and FTH1 decreased (P<0.01), while the mRNA and protein expression of ACSL4 increased (P<0.01). In comparison with the model group, the acupuncture and Fer-1 groups showed the increase of testicular wet weight and coefficient (P<0.01), sperm concentration and motility (P<0.01), and the serum contents of T, FSH, and LH (P<0.01); and the improvements in testicular and epididymal histopathology; ROS expression and the contents of MDA and TFe decreased (P<0.01), and the content of GSH elevated (P<0.05); the mitochondrial structure and numbers were ameliorated and mitochondrial membrane potential rose (P<0.01). Besides, in comparison with the model group, the mRNA expression of SLC7A11 was higher (P<0.05, P<0.01), the mRNA and protein expression of GPX4 and FTH1 increased (P<0.01, P<0.05), and the mRNA and protein expression of ACSL4 decreased (P<0.01) in the acupuncture and the Fer-1 groups; and the protein expression of SLC7A11 was higher in the Fer-1 group (P<0.05).
CONCLUSION
"Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture technique may improve the reproductive capacity in the mice with asthenospermia by alleviating ferroptosis-induced cellular damage and ameliorating testicular function.
Animals
;
Male
;
Ferroptosis
;
Mice
;
Acupuncture Therapy
;
Mice, Inbred C57BL
;
Asthenozoospermia/metabolism*
;
Humans
;
Acupuncture Points
;
Testis/metabolism*
;
Luteinizing Hormone/metabolism*
;
Malondialdehyde/metabolism*
;
Reproduction
;
Testosterone/metabolism*

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