Ferroptosis, a novel form of programmed cell death driven by iron-dependent lipid peroxidation, plays a crucial role in both disease treatment and microbial control due to its multi-level regulatory mechanisms. It mainly involves iron metabolism, lipid peroxidation, and antioxidant systems. In the field of disease treatment, ferroptosis is regarded as a highly promising therapeutic target because of its key role in autoimmune diseases, cancer, and cardiovascular diseases. This review systematically summarizes the core regulatory factors of ferroptosis, such as glutathione peroxidase 4 (GPX4) and long-chain acyl-CoA synthetase 4 (ACSL4) and their interaction networks, and deeply explores the application prospects of targeted intervention strategies based on ferroptosis signaling pathways in disease treatment and microbial control. Additionally, we also summarize the current issues faced by ferroptosis in practical applications and proposes strategies, such as nanodelivery, improved drug chemical stability and enhanced water solubility to optimize therapeutic efficacy. We aim to provide a theoretical basis and practical guidelines for exploring more targeted treatments using ferroptosis.

The emergence of CRISPR/Cas system has greatly promoted the progress in the field of gene editing, especially CRISPR/Cas9 system, which has become a core tool in biomedical research. Long noncoding RNAs (lncRNAs) play a key role in gene regulation, cell differentiation and the development of a variety of diseases. Especially in cancer research, lncRNAs have important application prospects as cancer biomarkers and therapeutic targets. However, lncRNAs are generally characterized by low abundance and poor conservation, which limits the study of their functions by traditional means. CRISPR/Cas9 technology provides an efficient, flexible and accurate tool for lncRNA research, which significantly accelerates the progress in this field. This paper first reviews the basic principles of CRISPR/Cas9 system and its wide applications in gene editing, including CRISPR knockout, knock-in, interference, activation and other functional systems. These technologies can not only screen key lncRNAs in specific biological processes, but also be used for gene function research to explore their roles in diseases. This article focuses on the analysis of CRISPR/Cas9 technology in the study of lncRNA functions, regulatory mechanisms, and its key applications in tumor research. In addition, the article also summarizes the methods of genome-wide screening by CRISPR/Cas9 to identify functional lncRNAs, and discusses the roles of these lncRNAs in cancer cell proliferation, migration, invasion and drug resistance. CRISPR/Cas9 knockout system can efficiently knock down lncRNA genes and reveal their specific functions in gene regulation. At the same time, CRISPR activation and interference technology provide a new idea for the research of noncoding genes, and further explore its clinical application in cancer and other diseases by regulating the expression level of lncRNAs. The article also discusses the potential of CRISPR technology in future lncRNA research, especially the progress in solving technical problems such as genome complexity, targeting efficiency and off-target effects. As mentioned in the review, CRISPR/Cas9 technology not only provides a powerful tool for studying lncRNAs, but also provides new ideas and opportunities for developing new means of cancer diagnosis and treatment in the future.

mRNA therapy involves delivering target molecules in the form of mRNA into cells to treat diseases. The highly variable nature of mRNA sequences offers potential solutions for high-throughput drug discovery and personalized treatment. This review begins with an overview of the development history of mRNA, tracing its journey from discovery to becoming a potential treatment. The review also discusses the applications of mRNA in protein replacement therapy, cancer treatment, in vivo gene editing, and infectious disease prevention based on the different categories of proteins delivered by mRNA. Additionally, optimizing mRNA formulations and their delivery vehicles is crucial for clinical application. This review further explains how to enhance the translation efficiency and stability of mRNA through nucleoside modifications and sequence optimization, and we systematically compare the pros and cons of novel circular mRNA versus traditional linear mRNA in vaccine development. Moreover, we summarize common delivery methods, such as lipid nanoparticles, and discuss the latest advancements in targeted delivery systems. For currently approved and in-development mRNA drugs, we systematically review the diseases treated, effector molecules delivered, and their clinical stages. Finally, we explore the challenges facing mRNA therapies and the potential diseases they could address, aiming to provide a theoretical foundation and reference for the development of mRNA therapies.

Gene editing technology has become an important tool for biomedical research because of its high efficiency and precision in target gene localization and cleavage. The technology not only facilitates basic research on gene function, but also provides new strategies for gene therapy of hereditary diseases and genetic improvement of crops. With the integration of artificial intelligence (AI) technology, especially the application of machine learning algorithms, the design and execution of gene editing have become more intelligent. AI technology optimizes the design of sgRNAs through predictive analysis and pattern recognition, improving the specificity and efficiency of editing while reducing the risk of off-target effects. In addition, AI plays a key role in the parsing of large-scale genomic data, providing new perspectives for understanding complex biological processes and disease mechanisms. This paper reviews the research progress of data-driven gene editing technology in target precision, safety enhancement and personalized therapy, aiming to provide reference and inspiration for researchers in the field of gene editing technology and to promote the application and development of AI in gene editing technology.

Ferroptosis is a new form of programmed cell death with iron-dependent lipid peroxidation as its core. A variety of metabolites are involved in the regulation of ferroptosis, among which lipid metabolism plays an important role. The most classic lipid metabolism-mediated ferroptosis mechanism is that phospholipids containing polyunsaturated fatty acyl chain (PUFA-PLs) located on biological membranes undergo super-threshold peroxidation, which leads to the destruction of membrane structure and function. In addition, special lipids containing polyunsaturated fatty acid (PUFA), such asphospholipid withdiacyl-PUFA tails (PL-PUFA₂), polyunsaturated ether phospholipid (PUFA-ePL), cholesterol ester containing polyunsaturated fatty acyl chain (PUFA-CE) have also been found to be involved in the ferroptosis process by providing PUFA for peroxidation. Lipid droplets was also found to regulate the sensitivity of ferroptosis through storing and releasing PUFA. Intermediates and derivatives of cholesterol metabolism are mainly involved in the negative regulation of ferroptosis, whereas different classes of sphingolipids were reported to have inconsistent regulatory directions for ferroptosis. A large number of previous studies have confirmed that ferroptosis is closely related to the metastasis and drug resistance of gastrointestinal tumors, therefore, we further summarized the lipid metabolism mechanisms related to ferroptosis resistance in gastrointestinal tumor cells, such as weakening the anabolism and peroxidation processes of PUFA-PLs, enhancing the ferroptosis defense system and so on. At the same time, we elaborated on the relationship between cholesterol metabolism, lipid drop metabolism, sphingolipid metabolism, and ferroptosis resistance in gastrointestinal tumors. Targeting these specific lipids, metabolic enzymes, and pathways to regulate ferroptosis has important clinical potential value. It is expected to provide new ideas for finding new diagnostic and prognostic markers, therapeutic drugs, and reversing chemotherapy resistance for gastrointestinal tumors.

Cardiolipin (CL) is a special type of polyglycerophospholipid, primarily synthesized in the mitochondrial inner membrane and cristae, and serves as a key component for mitochondrial function. It plays an essential role in the cellular membrane, mitochondrial inner membrane and energy metabolism, especially in maintaining the stability of oxidative phosphorylation and the electron transport chain. Abnormal metabolism of cardiolipin is closely associated with the occurrence of various cardiovascular diseases, particularly in genetic disorders such as Barth syndrome (BTHS). Moreover, the role of cardiolipin peroxides in cardiovascular diseases has been increasingly recognized. Studies have shown that cardiolipin peroxidation not only leads to damage of the mitochondrial inner membrane but also promotes the generation of reactive oxygen species (ROS), thereby enhancing oxidative stress within the cell. Abnormal metabolism of cardiolipin is also closely related to the pathogenesis of atherosclerosis, diabetic cardiomyopathy, hypertension, and other diseases. Regulating cardiolipin metabolism and repairing its functional defects may offer potential strategies for treating these diseases. This review discusses the synthesis, degradation, and remodeling processes of cardiolipin, and explores its significant role in cardiovascular diseases. The synthesis of cardiolipin relies on various enzymes within the mitochondria, while its remodeling involves key enzymes such as phosphatidyltransferases. Abnormal metabolism of cardiolipin, particularly the CL remodeling defects caused by tafazzin gene mutations in BTHS patients, leads to mitochondrial dysfunction, reduced ATP synthesis, increased oxidative stress, and ultimately results in myocardial and other tissue damage.

Aging is the result of the accumulation of various molecular and cellular damages over time, encompassing 12 characteristic hallmarks divided into three categories. These include primary hallmarks such as genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, and disabled macroautophagy; antagonistic hallmarks like deregulated nutrient sensing, mitochondrial dysfunction, and cellular senescence; and integrative hallmarks including stem cell exhaustion, altered intercellular communication, chronic inflammation and dysbiosis. Therefore, investigating cellular signaling factors within a single pathway is insufficient to comprehensively understand the complex mechanisms underlying aging. Casein kinase II (CK2), one of the earliest identified protein kinases, is capable of phosphorylating serine/threonine/tyrosine residues on hundreds of substrates. It exhibits highly constitutive expression and activity, and is extensively involved in the regulation of cellular processes such as proliferation, differentiation, apoptosis, stress response, metabolism, and immune function. CK2 plays a unique role in coordinating the cross-talk and integration of various signaling pathways, which is crucial for maintaining cell survival and homeostasis. Recent studies have revealed that CK2 exhibits perturbations in both expression levels and enzymatic activity during aging process, with notable heterogeneity observed across different animals, tissues and cellular models. Overall, the downregulation of CK2 expression not only promotes the development of primary hallmarks but also alleviates antagonistic hallmarks and ameliorates integrative hallmarks of aging, demonstrating dual effect and interconnected mechanisms. Notably, aberrant activation in CK2 expression and activity are associated with various aging-related diseases, including cancer, cardiovascular diseases, chronic metabolic disorders, neurodegenerative diseases and skeletal degenerative conditions. Therefore, maintaining CK2 homeostasis may represent an effective strategy for delaying aging. This review summarizes the latest advances in CK2 and aging research, providing not only deeper insights into the common mechanisms underlying aging and aging-related diseases, but also a theoretical foundation for identifying potential targets for the early prevention of aging-related diseases.

Skeletal muscle is the largest metabolic and endocrine organ in the body. As the basic unit of skeletal muscle, muscle fiber has high plasticity. Skeletal muscle fibers are mainly divided into oxidative and glycolytic muscle fibers, and muscle fibers type is an important factor affecting the contraction and energy metabolism of skeletal muscle. Understanding the molecular mechanism regulating skeletal muscle fiber type conversion is of great significance for regulating skeletal muscle-related diseases. Mitochondria is the energy factory of cell life activities, and the characteristics of mitochondria, namely the content, shape and distribution of mitochondria, are closely related to the function of mitochondria. The mitochondrial characteristics of various types of muscle fiber are different, which is related to the difference of energy metabolism of different muscle fiber types. Mitochondrial homeostasis is a dynamic equilibrium process, which is usually regulated by mitochondrial biogenesis, mitochondrial fusion and fission, mitochondrial autophagy and other processes. These processes not only affect the morphology and quantity of mitochondria, but also affect the metabolic balance of glucose and fatty acids in the body. Many studies have also shown that mitochondria-mediated changes in the substrates of energy metabolism of skeletal muscle affect the type conversion process of skeletal muscle fiber. Exercise is a non-drug treatment that can generally promote the formation of oxidized muscle fibers by maintaining skeletal muscle mitochondrial homeostasis. In this paper, the mitochondrial characteristics of different types of skeletal muscle fibers and the role of maintaining mitochondrial homeostasis in regulating muscle fiber type conversion were reviewed. On this basis, the molecular mechanism of mitochondrial involvement in PGC1α, Ca²⁺ and ROS signaling pathways mediated by fiber type conversion of skeletal muscle was summarized. Mitochondria are the energy factories of skeletal muscle, and targeted intervention through understanding their regulatory mechanism may be a new direction for the treatment of skeletal muscle-related diseases in the future.

The complement system plays an important role in identifying and eliminating pathogens, clearing physiological debris, coordinating immune response and homeostasis, etc. As an early warning signal of inflammatory response, abnormal complement activity is an important cause of pathologic pain diseases. Complement 3 (C3) is an important indicator of the activation of the complement system during pathologic pain induction. Experimental and clinical epidemiological studies have found abnormal increase of C3 in peripheral and central nerve in various types of pathologic pain, and C3 directly or indirectly regulates the response of neurons to neuropathy by binding specific C3 receptors. C3 can directly regulate various aspects of neuronal life and death through specific complement receptors expressed on the plasma membrane of neurons, and can also indirectly regulate by recruiting glial cells and immune cells through various mechanisms to transmit complement signals to neurons, which guide the response of neurons to tissue injury, neurotrauma, and neuropathy. This article mainly reviews the mechanism of cytokines and signaling pathways involved in C3 in pathologic pain, and discusses the possibility of C3 as an analgesic target in pathologic pain.

Protein acylation is a group of recently identified chemical modification closely linked to metabolism with the metabolic intermediate acyl-coenzyme A (acyl-coA) as the substrate. Acylation possesses similar chemical structures to acetylation, with differences in carbon chain length, hydrophobicity, and charge. Most acyl-CoAs are intermediate metabolites, the level of which are influenced by the intracellular metabolic state. Therefore, protein acylation is greatly affected by cellular metabolism. Metabolic reprogramming is an important feature of tumor cells. In addition to the classic "Warburg effect", cancer cells exhibit abnormal regulation in lipid metabolism, amino acid metabolism and biological oxidation. Acylation on histones can impact chromatin structure, regulating gene expression and DNA repair critically involved in cancer progression. On the other hand, acylation on non-histones can regulate signal transduction, enzymatic activity, or protein-protein interactions to affect cancer cell behaviors such as proliferation, invasion, immune evasion, and vascular remodeling. Focusing on three types of acylation closely related to metabolism: lactylation, succinylation, and crotonylation, this article introduces the production, raw materials, regulatory mechanism and factors of protein acylation. We then review representative studies to show how cancer cell metabolic reprogramming can regulate these processes and histone/non-histone acylation levels, which subsequently affect the expression and function metabolism-related genes/proteins to form a bidirectional dialogue and accelerate cancer progression. In addition, we present several prospects for potential research directions and translational applications in the field.

Currently, acquired immune deficiency syndrome (AIDS) has emerged as a global public health crisis that profoundly compromises human immune defenses. By systematically dismantling the immune system, human immunodeficiency virus (HIV) renders individuals vulnerable to opportunistic infections and malignancies, ultimately culminating in AIDS progression. It is urgent to eradicate the latent HIV virus and achieve a functional cure, thus limiting the development of AIDS and improving the quality of patients. Epigenetics investigates heritable alterations in gene expression that occur independently of DNA sequence. The intricate regulation of HIV gene expression is orchestrated through multifaceted epigenetic mechanisms involving both viral and host factors. Understanding the epigenetic mechanisms associated with HIV infection is crucial for clearing latent viruses and achieving control and treatment of AIDS in the future. Therefore, we will discuss the epigenetic regulatory patterns and mechanisms involved in HIV infection, particularly emphasize on four principal mechanisms: DNA methylation, histone modification, non-coding RNA regulation and RNA modification. We comprehensively analyze how these regulatory factors influence the viral life cycle, particularly regarding latency establishment, reactivation dynamics, and persistent infection maintenance. Furthermore, we delineate the interplay between epigenetic regulators and key cellular signaling pathways during HIV pathogenesis. The review culminates in a critical appraisal of recent breakthroughs and persistent challenges in epigenetics-based therapeutic strategies, while highlighting innovative approaches for functional cure development. By elucidating the pivotal role of epigenetic regulation in HIV latency, this review aims to establish a novel theoretical foundation and innovative research directions for next-generation AIDS therapeutics rooted in epigenetic modification.

Plant-derived exosome-like nanovesicles refer to spherical lipid bilayer vesicles isolated from plants that contain lipids, proteins, RNAs, and various small molecules. These nanovesicles exhibit diverse biological activities, including anti-inflammatory, anti-tumor, antioxidant, and drug delivery properties. However, the functional characteristics of nanovesicles derived from rhizoma polygonati remain unexplored. In this study, exosome-like nanovesicles derived from rhizoma polygonati (referred to as RP-EVs) were successfully isolated using ultracentrifugation and density gradient centrifugation. Their physicochemical properties and anti-inflammatory functions were systematically characterized. Our results show that RP-EVs are predominantly negatively charged, with an average particle size of 166.5 ± 3.3 nm, and are spherical lipid vesicles. Cellular uptake assays demonstrated that RP-EVs can be phagocytized by macrophages. qPCR and ELISA experiments revealed that RP-EVs can inhibit the elevation of interleukin 6 (IL-6), interleukin 1β (IL-1β), and tumor necrosis factor-alpha (TNF-α) induced by lipopolysaccharide (LPS) stimulation (^****P < 0.0001). Additionally, reactive oxygen species (ROS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assays confirmed that RP-EVs exhibit antioxidant properties (^*P < 0.05). Further investigation of the underlying mechanisms through immunofluorescence and Western blotting revealed that RP-EVs inhibit the nuclear translocation (^**P < 0.01) and phosphorylation (^***P < 0.001) of nuclear factor kappa-B p65 (NF-κB p65) via the IκBα/NF-κB signaling pathway, thereby regulating the expression of inflammatory mediators. In animal experiments, intraperitoneal injection of RP-EVs into mice for 48 hours showed predominant localization in the liver and spleen. Finally, an acute inflammatory mouse model was established via intraperitoneal injection of LPS. qPCR and ELISA analyses demonstrated that RP-EVs alleviated the expression of inflammatory factors in both the serum and spleen of LPS-treated mice (^*P < 0.05). In conclusion, this study isolated RP-EVs and elucidated their anti-inflammatory properties and potential mechanisms. These findings provide valuable insights into the functional exploration of nanoparticle vesicles derived from traditional Chinese medicine and suggest a novel therapeutic strategy for the treatment of inflammation-related diseases.

The structure and function of mitochondria and endoplasmic reticulum (ER) are important for maintaining cellular homeostasis. It has been found that the interaction between mitochondria and ER is involved in the occurrence and development of a variety of diseases. The mitochondria-associated ER membrane (MAM) is a membrane contact site between the ER and mitochondria, and is an important communication center between organelles in eukaryotic cell. Calcium channels on the ER side and the mitochondrial side are crucial in the calcium transport process in MAM. The interaction between ER and mitochondria controls mitochondrial biological function and cell survival through calcium transport regulation, and are involved in the occurrence and development of various pathologic process. On the one hand, MAM regulates calcium transport, which is involved in the modulation of various cellular survival and death processes. It plays a profound regulatory role in the damage of tumor cells, neuronal cells, cardiomyocytes, endothelial cells and nucleus pulposus cells through different key molecules within MAM.On the other hand, the regulation of MAM in calcium transport is crucial in the development of mitochondrial dysfunction in Hepa 1-6 cells, the synthesis and secretion of pancreatic β-cells and amyotrophic lateral sclerosis. In addition, MAM also affects cellular transcription processes by regulating calcium transport, thereby exerting significant regulatory effects on angiogenesis and breast cancer. This paper reviews the structural features and pathophysiologic role of calcium transport regulation of MAM, and expects to provide new horizons for prevention and treatment of related diseases targeting MAM.

In the canonical model of gene transcription, transcription factors regulate the transcription of target genes by binding to double-stranded DNA (dsDNA) containing its specific consensus motifs. In contrast to dsDNA, G-quadruplexes are atypical nucleic acid secondary structures formed by guanine-rich sequences. G-quadruplex structures are involved in regulating various biological processes, including gene transcription, and have emerged as a significant research topic in the field of molecular biology. Recently, many research groups, including us, have revealed that G-quadruplex structures recruit transcription factors to bind to the promoters more efficiently than dsDNA, thereby enhancing target gene expression. However, there is currently a lack of comprehensive summaries and discussions regarding this atypical model of gene transcription regulation. In this review, we first elucidate the characteristics of G-quadruplex structures and the techniques used to identify these structures. G-quadruplexes can be classified into intramolecular and intermolecular types; intramolecular G-quadruplexes are further divided into parallel, antiparallel and hybrid types. The G-quadruplex structures can be determined using techniques such as circular dichroism, nuclear magnetic resonance spectroscopy and gel migration, among others. Then, we discuss the regulatory role of G-quadruplex in gene transcription. G-quadruplexes are mainly highly enriched in gene promoter. Early studies revealed that G-quadruplexes can inhibit gene transcription. However, lots of studies have recently proven that this structure has a new function of recruiting transcription factors to activate gene transcription. Finally, we summarize the classification of transcription factors with G-quadruplex binding activity, including transcription factors with C₂H₂ zinc fingers, forked head/winged helices, and p53 domains. The DNA-binding domain determines the interaction between transcription factors and G-quadruplex. Moreover, we propose future research directions in the field of G-quadruplex and transcription factors in regulating gene transcription. In conclusion, this review can provide important guidance for understanding the concept of G-quadruplex as a cis-acting element for transcriptional activation.

Liquid-liquid phase separation (LLPS) is the process where intracellular biomolecules rapidly form reversible high concentrations of liquid-phase condensates, resulting in the compartmentalization and the formation of intracellular membraneless organelles. LLPS is involved in various biological processes and pathologic processes, such as neurodegenerative diseases and cancers. Long noncoding RNAs (lncRNAs) have been revealed to be closely related to phase separation. It has become a research hotspot in life science recently, because it provides new insight into the mechanism of lncRNAs. In this review, we focuses on the effect of lncRNA SLERT on the phase separation of nucleolar fibrillar center/dense fibrillar component (FC/DFC) by interacting with DDX21 protein as a molecular chaperone; LINC00657 (NORAD) forms NP bodies with PUM proteins, which drives PUM protein liquid droplets and inhibits its activity, and promotes genomic stability; DilncRNA regulates DNA damage response small RNAs (DDRNAs) and LLPS of p53 binding protein 1 (53BP1) in response to DNA damage, and lncRNA LINP1 phase separation droplets bind to Ku protein to promote DNA damage repair; LncRNA SNHG9, MELTF-AS1 and MALR drive LATS1, YBX1 and ILF3 protein LLPS respectively to promote cancer, while GIRGL and lncFASA act as tumor suppressor genes in cancer development through regulating the phase separation of CAPRIN1 and PRDX1 respectively; LncRNA XIST drives X chromosome inactivation by LLPS. In a word, we summarize the latest research progress about the functional roles of lncRNAs in FC/DFC nucleolus, genomic stability and DNA damage and repair, cancer and X-chromosome inactivation through regulating LLPS. This paper shows that lncRNAs can participate in multiple pathophysiological processes by regulating LLPS, which is expected to provide a new direction for the treatment of LLPS-mediated diseases.

With the continuous increase in global obesity prevalence, the impact of obesity on reproductive physiology has garnered widespread societal attention. As a metabolic disorder, obesity is typically accompanied by multiple abnormal physiological phenomena, such as excessive adipose accumulation and exacerbated inflammatory responses, which severely compromise the reproductive health of humans and animals. Reproductive damage induced by obesity involves a series of complex biochemical reactions and in vivo metabolic pathways, manifesting as impaired male sperm quality and female fertility. To better understand the relationship between obesity and reproductive physiology, this review summarizes the reproductive injuries caused by obesity and their underlying mechanisms. In the obese state, conditions such as oxidative stress, insulin resistance, and hyperinsulinemia are induced, with adipokines (leptin, adiponectin, resistin, etc.) and inflammatory factors (TNF-α, IL-6, IL-1β, etc.) interacting synergistically to affect the reproductive system. Oxidative stress activates the MAPK and NF-κB pathways, interfering with insulin signaling, while chronic inflammation leads to adipocyte secretory disorders and disrupts the hypothalamic-pituitary-gonadal regulatory axis. Studies have shown that obese males exhibit significantly decreased testosterone levels and impaired sperm quality, whereas obese females suffer from reproductive hormone imbalance, ovulation disorders, and polycystic ovary syndrome. This review discusses how obesity-induced metabolic disorders lead to impaired reproductive physiology in both males and females, along with the underlying mechanisms, providing a theoretical basis for the prevention and treatment of obesity-related reproductive disorders in the future.

The incidence of non-alcoholic fatty liver disease (NAFLD) has been increasing annually. Current primary treatment strategies involve dietary modifications and increased physical activity to alleviate symptoms, yet there is a notable lack of targeted pharmacological interventions. Members of the micro RNA-29 (miR-29) family (miR-29a, miR-29b, miR-29c) are known to play a critical regulatory role in lipid metabolism within hepatocytes; however, the underlying mechanisms remain to be elucidated. This study aims to identify the target genes and associated signaling pathways of the miR-29 family, thereby providing potential therapeutic targets for the development of NAFLD treatments. Firstly, the human liver cell line HepG2 was utilized as a model for adipogenic induction, and miR-29a/b/c-3p mimics were individually transfected. Through methods such as Oil Red O staining and triglyceride (TG) quantification, it was observed that the miR-29 family members significantly inhibited lipid accumulation in hepatocytes (P<0.05). Subsequently, qRT-PCR and Western blot were utilized to detect the expression levels of adipogenic marker genes (fatty acid synthase (FAS), acetyl coa carboxylase (ACACA) , stearoyl-coenzyme a desaturase1 (Scd1)) and autophagy marker genes (sequestosome 1 (SQSTM1, also known as p62), autophagy related gene 5 (Atg5)), and the results indicated that the members of the miR-29 family could significantly suppress the expression of FAS, ACACA, Scd1, and p62 genes in hepatocytes, while significantly enhancing the level of the Atg5 gene. Further investigations using signaling pathway activity analysis and dual luciferase reporter assays confirmed that the miR-29a/b/c could suppress the mTOR signaling pathway activity and directly interact with the ten-eleven translocation 2 (TET2) gene. Finally, co-transfection experiments were performed to examine the potential synergistic effects among the miR-29-3p family members, and the results demonstrated that co-transfection of miR-29 family members more effectively inhibited lipid droplet accumulation in HepG2 cells and further suppressed the expression of the target gene TET2 compared to individual transfection. In summary, the miR-29 family members may reduce lipid accumulation in hepatocytes by inhibiting the mTOR signaling pathway via the TET2 gene, and they exhibit a positive synergistic effect.