Nuclear Deformation Research Articles

Mechanical memory based on chromatin and metabolism remodeling promotes proliferation and smooth muscle differentiation in mesenchymal stem cells.

Stem cells respond and remember mechanical cues from the microenvironment, which modulates their therapeutic effects. Chromatin organization and energy metabolism regulate the stem cell fate induced by mechanical cues. However, the mechanism of mechanical memory is still unclear. This study aimed to investigate the effects of mechanical amplitude, frequency, duration, and stretch cycle on mechanical memory in mesenchymal stem cells. It showed that the amplitude was the dominant parameter to the persistence of cell alignment. F-actin, paxillin, and nuclear deformation are more prone to be remolded than cell alignment. Stretching induces transcriptional memory, resulting in greater transcription upon subsequent reloading. Cell metabolism displays mechanical memory with sustained mitochondrial fusion and increased ATP production. The mechanical memory of chromatin condensation is mediated by histone H3 lysine 27 trimethylation, leading to much higher smooth muscle differentiation efficiency. Interestingly, mechanical memory can be transmitted based on direct cell-cell interaction, and stretched cells can remodel the metabolic homeostasis of static cells. Our results provide insight into the underlying mechanism of mechanical memory and its potential benefits for stem cell therapy.

Targeting SRSF2 mutations in leukemia with RKI-1447: A strategy to impair cellular division and nuclear structure

Spliceosome machinery mutations are common early mutations in myeloid malignancies; however, effective targeted therapies against them are still lacking. In the current study, we used an invitro high-throughput drug screen among four different isogenic cell lines and identified RKI-1447, a Rho-associated protein kinase inhibitor, as selective cytotoxic effector of SRSF2 mutant cells. RKI-1447 targeted SRSF2 mutated primary human samples in xenografts models. RKI-1447 induced mitotic catastrophe and induced major reorganization of the microtubule system and severe nuclear deformation. Transmission electron microscopy and 3D light microscopy revealed that SRSF2 mutations induce deep nuclear indentation and segmentation that are apparently driven by microtubule-rich cytoplasmic intrusions, which are exacerbated by RKI-1447. The severe nuclear deformation in RKI-1447-treated SRSF2 mutant cells prevents cells from completing mitosis. These findings shed new light on the interplay between microtubules and the nucleus and offers new ways for targeting pre-leukemic SRSF2 mutant cells.

Atypical cofilin signaling drives dendritic cell migration through the extracellular matrix via nuclear deformation

To mount an adaptive immune response, dendritic cells must migrate to lymph nodes to present antigens to Tcells. Critical to 3D migration is the nucleus, which is the size-limiting barrier for migration through the extracellular matrix. Here, we show that inflammatory activation of dendritic cells leads to the nucleus becoming spherically deformed and enables dendritic cells to overcome the typical 2- to 3-μm diameter limit for 3D migration through gaps in the extracellular matrix. We show that the nuclear shape change is partially attained through reduced cell adhesion, whereas improved 3D migration is achieved through reprogramming of the actin cytoskeleton. Specifically, our data point to a model whereby the phosphorylation of cofilin-1 at serine 41 drives the assembly of a cofilin-actomyosin ring proximal to the nucleus and enhances migration through 3D collagen gels. In summary, these data describe signaling events through which dendritic cells deform their nucleus and enhance their migratory capacity.

A Microfluidic Perfusion Culture Setup to Investigate Cell Migration in 3D Constrictions

AbstractCell migration is a fundamental process underlying the morphological maturation of organs, but also in disease‐related conditions such as cancer. Cells are able to migrate through crowded space and a tight extracellular matrix (ECM). Passing through a constriction, a cell deforms strongly, including its nucleus. Such nuclear deformation can lead to changes in the 3D‐genomic architecture, and putatively, DNA methylation. However, the specific effects of deformation on cells are not well understood. It is highly desired to establish an ex vivo methodology to induce well‐defined cell deformation in complex geometrical constrictions. This study introduces a microfluidic system for the study of migrating cells in precisely controlled geometrical confinement. A procedure for coating, seeding of cerebellar granule cells, and perfusion culture is presented. By leveraging direct laser writing, channels with smooth, anisotropically curved surfaces on the cell‐scale can be fabricated. The system consists of constriction channels with a radius of 2 or 4 µm for the cells to pass through. This corresponds to a compression of the nucleus to 3.5% and 14.2% of its undeformed cross‐sectional area, respectively. The system can be used to investigate the influence of confinement geometry on the migration behavior and transcriptome of various cell types.

Generic multi-particle transverse momentum correlations as a new tool for studying nuclear structure at the energy frontier

The mean transverse momentum of produced particles, [pT]\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$[p_\ extrm{T} ]$$\\end{document}, and its event-by-event fluctuations give direct access to the initial conditions of ultra-relativistic heavy-ion collisions and help probe the colliding nuclei’s structure. The [pT]\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$[p_\ extrm{T} ]$$\\end{document} fluctuations can be studied via multi-particle pT\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$p_\ extrm{T}$$\\end{document} correlations; so far, only the lowest four orders have been studied. Higher-order fluctuations can provide stronger constraints on the initial conditions and improved sensitivity to the detailed nuclear structure; however, their direct implementation can be challenging and is still lacking. In this paper, we apply a generic recursive algorithm for the genuine multi-particle pT\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$p_\ extrm{T}$$\\end{document} correlations, which enables the accurate study of higher-order [pT]\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$[p_\ extrm{T} ]$$\\end{document} fluctuations without heavy computational cost for the first time. With this algorithm, we will examine the power of multi-particle pT\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$p_\ extrm{T}$$\\end{document} correlations through Monte Carlo model studies with different nuclear structures. The impact on the nuclear structure studies, including the nuclear deformation and triaxial structure, will be discussed. These results demonstrate the usefulness of multi-particle pT\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$p_\ extrm{T}$$\\end{document} correlations for studying nuclear structure in high-energy nuclei collisions at RHIC and the LHC, which could serve as a complementary tool to existing low-energy nuclear structure studies.

Prediction of novel effects in rotational nuclei at high speed

The study of high-speed rotating matter is a crucial research topic in physics due to the emergence of novel phenomena. In this letter, we combined cranking covariant density functional theory (CDFT) with a similarity renormalization group approach to decompose the Hamiltonian from the cranking CDFT into different Hermite components, including the non-relativistic term, the dynamical term, the spin-orbit coupling, and the Darwin term. Especially, we obtained the rotational term, the term relating to Zeeman-like effect, and the spin-rotation coupling due to consideration of rotation and spatial component of vector potential. By exploring these operators, we aim to identify novel phenomena that may occur in rotating nuclei. Signature splitting, Zeeman-like effect, and spin-rotation coupling are among the potential novelties that may arise in rotating nuclei. Additionally, we investigated the observability of these phenomena and their dependence on various factors such as nuclear deformation, rotational angular velocity, and magnetic field-like strength.

Nuclear mechanics and deformation regulate cellular lineage and senescence via YAP mechanotransduction

Nuclear mechanics and deformation regulate cellular lineage and senescence via YAP mechanotransduction

Hot Topics and Emerging Trends in Mechanobiology Research

Mechanobiology focuses on a series of important physiopathological processes, such as how cells perceive different mechanomechanical stimuli, the process of intracellular mechanotransduction, and how mechanical signals determine the behavior and fate of cells. From the initial stage of embryogenesis, to developmental biology and regenerative medicine, or even through the whole life process, mechanical signaling cascades and cellular mechanical responses in mechanobiology are of great significance in biomedical research. In recent years, research in the field of mechanobiology has undergone remarkable development. Several scientific consortia around the world have been analyzing mechanobiological processes from different perspectives, aiming to gain insights into the regulatory mechanisms by which mechanical factors affect cell fate determination. In this article, we summarized and reviewed the topics that have attracted more research interests in recent years in the field of mechanobiology, for example, arterial blood vessels, stem cell, and ion channel. We also discussed the potential trends that may emerge, such as nuclear deformation, fibrous extracellular matrix, tumor mechanobiology, cellular mechanotransduction, and piezo ion channels. In addition, we put forward new ideas concerning the limitations of mechanism research and the importance of big data analysis and mining in this field, thereby providing objective support and a systematic framework for grasping the hot research topics and exploring new research directions in the field of mechanobiology.

ATLAS measurement of mean momentum fluctuations and correlations with the flow in Xe+Xe and Pb+Pb collisions

The thermal fluctuations in the QGP medium formed in heavy ion collisions present themselves as event-wise mean transverse momentum ([pT]) fluctuations in the final state. Recent studies have shown that the fluctuations are sensitive to radial flow, random thermal motion, and nuclear deformation. They can provide constraints on the extent of thermalization of the QGP droplet. Recently, correlations of [pT] with flow harmonic coefficients (vn) also gained interest, as they were found to be sensitive primarily to the initial state fluctuations. This talk presents precise measurements of up to 3rd order [pT] cumulants as a function of multiplicity and centrality with an emphasis on ultra-central collisions. These results have strong implications for understanding the impacts of the initial condition, medium thermalization, and medium properties on final state [pT] fluctuations.

Quantitative analysis of nuclear deformations and DNA damage foci dynamics by live-cell imaging.

The correct repair of DNA Double Strand Breaks (DSBs) is fundamental to prevent the loss of genetic information, mutations, and chromosome rearrangements. An emerging determinant of DNA repair is chromatin mobility. However, how chromatin mobility can influence DSBs repair is still poorly understood. While increased mobility is generally associated with the correct repair by Homologous Recombination (HR) of DSBs generated in heterochromatin, it promotes the mis-repair of multiple distal DSBs by Non-Homologous End Joining (NHEJ). Here we describe a method for detecting and quantifying DSBs mobility by live-cell imaging in the context of multiple DSBs prone to mis-repair by NHEJ. In addition, we discuss a set of parameters that can be used for quantitative and qualitative analysis of nuclear deformations and to discard nuclei where the deformation could affect the analysis of DSBs mobility. While this method is based on the visualization of DSBs with the mCherry-53BP1-2 fusion protein, we believe that it can also be used to analyze the mobility of nuclear foci formed by different fluorescent proteins.

Neutron-rich clustering investigation at LNS with FARCOS detectors

The physics of clusters in heavy neutron-rich ions is a topic of constant interest in the worldwide scientific community. In recent times, many interesting phenomena have been investigated especially thanks to the development of new radioactive beam production facilities. One example is the clustering of α particles in neutron-rich isotopes of self-conjugated nuclei, such as 10Be or 16C, exhibiting even very large nuclear deformations. At Laboratori Nazionali del Sud of INFN, a study was carried out on the topic of α clustering, employing the CHIMERA and FARCOS detectors. Radioactive ions of interest, such as 10Be, 13B and 16C, were produced in a cocktail beam through the In-Flight fragmentation technique by the FRIBs@LNS facility. Particularly important for this study was the employment of four FARCOS detectors, offering high angular and energetic resolutions. Several calibration and analysis techniques have been exploited and developed in this experiment for the analysis of the data collected by FARCOS. Finally, some preliminary results on the analysis of 10Be and 16C spectroscopy will be shown, in relation to some results already collected in the literature.

Coupling of Perinuclear Actin Cap and Nuclear Mechanics in Regulating Flow-Induced Yap Spatiotemporal Nucleocytoplasmic Transport.

Mechanical forces, including flow shear stress, govern fundamental cellular processes by modulating nucleocytoplasmic transport of transcription factors like Yes-associated Protein (YAP). However, the underlying mechanical mechanism remains elusive. In this study, it is reported that unidirectional flow induces biphasic YAP transport with initial nuclear import, followed by nuclear export as actin cap formation and nuclear stiffening. Conversely, pathological oscillatory flow induces slight actin cap formation, nuclear softening, and sustained YAP nuclear localization. To elucidate the disparately YAP spatiotemporal distribution, a 3D mechanochemical model is developed, which integrates flow sensing, cytoskeleton organization, nucleus mechanotransduction, and YAP transport. The results unveiled that despite the significant localized nuclear stress imposed by the actin cap, its inherent stiffness counteracts the dispersed contractile stress exerted by conventional fibers on the nuclear membrane. Moreover, alterations in nuclear stiffness synergistically regulate nuclear deformation, thereby governing YAP transport. Furthermore, by expanding the single-cell model to a collective vertex framework, it is revealed that the irregularities in actin cap formation within individual cells have the potential to induce topological defects and spatially heterogeneous YAP distribution in the cellular monolayer. This work unveils a unified mechanism of flow-induced nucleocytoplasmic transport, providing a linkage between transcription factor localization and mechanical stimulation.

Exocrine gland-resident memory CD8+ T cells use mechanosensing for tissue surveillance.

Tissue-resident CD8+ T cells (TRM) continuously scan peptide-MHC (pMHC) complexes in their organ of residence to intercept microbial invaders. Recent data showed that TRM lodged in exocrine glands scan tissue in the absence of any chemoattractant or adhesion receptor signaling, thus bypassing the requirement for canonical migration-promoting factors. The signals eliciting this noncanonical motility and its relevance for organ surveillance have remained unknown. Using mouse models of viral infections, we report that exocrine gland TRM autonomously generated front-to-back F-actin flow for locomotion, accompanied by high cortical actomyosin contractility, and leading-edge bleb formation. The distinctive mode of exocrine gland TRM locomotion was triggered by sensing physical confinement and was closely correlated with nuclear deformation, which acts as a mechanosensor via an arachidonic acid and Ca2+ signaling pathway. By contrast, naïve CD8+ T cells or TRM surveilling microbe-exposed epithelial barriers did not show mechanosensing capacity. Inhibition of nuclear mechanosensing disrupted exocrine gland TRM scanning and impaired their ability to intercept target cells. These findings indicate that confinement is sufficient to elicit autonomous T cell surveillance in glands with restricted chemokine expression and constitutes a scanning strategy that complements chemosensing-dependent migration.

The Functions of Pt-DIC and Pt-Lamin B in Spermatogenesis of Portunus trituberculatus.

Cytoplasmic Dynein is a multiple-subunit macromolecular motor protein involved in the transport process of cells. The Dynein intermediate chain (DIC) is one of the subunits of Dynein-1. In our previous studies, we showed that Pt-DIC may play an important role in the nuclear deformation of spermiogenesis in Portunus trituberculatus. Lamin B is essential for maintaining nuclear structure and functions. Surprisingly, Pt-Lamin B was expressed not only in the perinucleus but also in the pro-acrosome during spermiogenesis in P. trituberculatus. Studies have also shown that Dynein-1 can mediate the transport of Lamin B in mammals. Thus, to study the relationship of Pt-DIC and Pt-Lamin B in the spermatogenesis of P. trituberculatus, we knocked down the Pt-DIC gene in P. trituberculatus by RNAi. The results showed that the distribution of Pt-DIC and Pt-Lamin B in spermiogenesis was abnormal, and the colocalization was weakened. Moreover, we verified the interaction of Pt-DIC and Pt-Lamin B via coimmunoprecipitation. Therefore, our results suggested that both Pt-DIC and Pt-Lamin B were involved in the spermatogenesis of P. trituberculatus, and one of the functions of Dynein-1 is to mediate the transport of Lamin B in the spermiogenesis of P. trituberculatus.

Many-body correlations for nuclear physics across scales: from nuclei to quark-gluon plasmas to hadron distributions

It is an experimental fact that multi-particle correlations in the final states of high-energy nucleus-nucleus collisions are sensitive to collective correlations of nucleons in the wave functions of the colliding nuclei. Here, I show that this connection is more direct than it intuitively seems. With an energy deposition scheme inspired by high-energy quantum chromodynamics, and within a linearized description of initial-state fluctuations in the quark-gluon plasma, I exhibit relations between N-particle correlations in the final states of nuclear collisions and N-nucleon density distributions in the colliding nuclei. This result formally justifies the sensitivity of the outcome of high-energy collisions to features such as nuclear deformations. It paves the way, thus, to systematic studies of the impact of state-of-the-art nuclear interactions in such processes.

Actin contraction controls nuclear blebbing and rupture independent of actin confinement.

The nucleus is a mechanically stable compartment of the cell that contains the genome and performs many essential functions. Nuclear mechanical components chromatin and lamins maintain nuclear shape, compartmentalization, and function by resisting antagonistic actin contraction and confinement. Studies have yet to compare chromatin and lamins perturbations side-by-side as well as modulated actin contraction while holding confinement constant. To accomplish this, we used nuclear localization signal green fluorescent protein to measure nuclear shape and rupture in live cells with chromatin and lamin perturbations. We then modulated actin contraction while maintaining actin confinement measured by nuclear height. Wild type, chromatin decompaction, and lamin B1 null present bleb-based nuclear deformations and ruptures dependent on actin contraction and independent of actin confinement. Actin contraction inhibition by Y27632 decreased nuclear blebbing and ruptures while activation by CN03 increased rupture frequency. Lamin A/C null results in overall abnormal shape also reliant on actin contraction, but similar blebs and ruptures as wild type. Increased DNA damage is caused by nuclear blebbing or abnormal shape which can be relieved by inhibition of actin contraction which rescues nuclear shape and decreases DNA damage levels in all perturbations. Thus, actin contraction drives nuclear blebbing, bleb-based ruptures, and abnormal shape independent of changes in actin confinement.

Competing failure modes in void and inclusion NiCoCrFeCu alloy under tensile simulation using molecular dynamic

This study exhibits dual effects from the parallel existence of inclusion and void on mechanical characteristics and deformation response for NiCoCrFeCu alloy based on an MD simulation of a tensile test. The pure Cu-inclusion and the defect rate 1.0 reinforce the strength of NiCoCrFeCu HEA, while pure void causes damage quickly of HEA. The nuclear deformation forms at the void root and boundary between the inclusion and workpiece for defect samples, which occurs in random zones for monolithic pieces. The deformation density for the sample combined void and inclusion is always the lowest because the boundary prevents the fusion between the hole-induced and workpiece-induced deformation. The defect rates are a critical factor in deciding the phase transition and deformation development during the strain stages, and the defect rate of 1.0 is the ladder. The tensile strength and strain energy increase as the defect rates increase from 0.43 to 1, then enormously decrease as the defect rate to 1.5, while Young’s modulus declines with an increase in defect rate. Moreover, the results also show that the effect of the void position and the divided number of voids with the same void volume in each material sample is severe. The tensile strength and Young’s modulus increase with the increase of the void number; the model with two voids in the X direction is subjected to higher stress than those with two holes in the Y direction. Finally, the mechanical parameters, such as the tensile strength and Young’s modulus, all decrease as the void radius and inclusion radius increase, and the sample can enhance elongation when adding an inclusion with a radius of 30 Å. The occupation of the twin boundary interferes with the development of dislocation, reducing the total dislocation length.

Study of octupole deformations in Pb-Pb collisions at 5.02 TeV

In this letter, we present the study of the role of octupole deformation in non-spherical nuclei in most-central Pb-Pb collisions at LHC energy regime. The sensitivity of octupole deformation β3 to the QGP observables is presented by employing Monte-Carlo HYDJET++ model. Motivated by the discrepancies in the v2-to-v3 puzzle found in Pb-Pb collisions and the low-energy nuclear structure calculations of nuclear deformation, we studied the first basic observables necessary for any study in heavy-ion collisions. Using HYDJET++ framework, we calculate the pseudorapidity distribution, transverse momentum (pT) spectra and average anisotropic flow (v2 and v3) of primary charged hadrons with different parameters in two geometrical configurations: body-body and tip-tip type of Pb-Pb collisions. The kinematic ranges 0<pT<20 GeV/c and |η|<0.8 are considered. We observe that the charged hadron multiplicity and transverse momentum spectra are dependent on the strength of octupole deformation parameter. 〈v2〉 and 〈v3〉 in body-body collisions show weak positive correlation with β3 while average anisotropic flow in tip-tip collisions are weakly correlated to β3 in most-central collision region.

Rare isotope-containing diamond colour centres for fundamental symmetry tests.

Detecting a non-zero electric dipole moment in a particle would unambiguously signify physics beyond the Standard Model. A potential pathway towards this is the detection of a nuclear Schiff moment, the magnitude of which is enhanced by the presence of nuclear octupole deformation. However, due to the low production rate of isotopes featuring such 'pear-shaped' nuclei, capturing, detecting and manipulating them efficiently is a crucial prerequisite. Incorporating them into synthetic diamond optical crystals can produce defects with defined, molecule-like structures and isolated electronic states within the diamond band gap, increasing capture efficiency, enabling repeated probing of even a single atom and producing narrow optical linewidths. In this study, we used density functional theory to investigate the formation, structure and electronic properties of crystal defects in diamond containing [Formula: see text], a rare isotope that is predicted to have an exceptionally strong nuclear octupole deformation. In addition, we identified and studied stable lanthanide-containing defects with similar electronic structures as non-radioactive proxies to aid in experimental methods. Our findings hold promise for the existence of such defects and can contribute to the development of a quantum information processing-inspired toolbox of techniques for studying rare isotopes. This article is part of the Theo Murphy meeting issue 'Diamond for quantum applications'.

A UPLC-QTOF/MS-based hepatic tissue metabolomics approach deciphers the mechanism of Huachansu tablets-based intervention against hepatocellular carcinoma

Huachansu (HCS) tablets, classified as well-known traditional Chinese medicine (TCM) preparation, have been proved to be effective in the treatment of hepatocellular carcinoma (HCC) in clinical studies. However, the underlying mechanism of HCS tablets against HCC has not been comprehensively elucidated. In this study, a rat model of HCC was established with diethylnitrosamine (DEN) inducer. The efficacy of HCS tablets against HCC was assessed through liver histopathological examination and evaluation of biochemical indicators. A metabolomics method based on UPLC-Q-TOF/MS combined with multivariate data analysis was established to identify differential metabolites related to the inhibition effect of HCS tablets on HCC, and then the relevant metabolic pathway analysis was performed to investigate the anti-HCC mechanisms of HCS tablets. The results showed that compared to the control group, the HCC model group showed a significant increase in the values of HCC-related biochemical indicators and the number of tumor nodules, indicating the successful establishment of the HCC rat model. Upon treatment with HCS tablets, the values of HCC-related biochemical indicators decreased, liver fibrosis and nuclear deformation were also significantly alleviated. A total of 15 differential metabolites associated with the anti-tumor effect of HCS tablets on HCC were screened and annotated through hepatic tissue metabolomics studies. Analysis of metabolic pathways revealed that the therapeutic effects of HCS tablets on HCC mainly involved the pentose and glucuronate interconversions and arachidonic acid metabolism. Further western blotting corroborated that the alteration in arachidonic acid (AA) level after the intervention of HCS tablets was related to the inhibition of cPLA2α expression in rat liver tissues. In conclusion, HCS tablets exhibit a certain anti-tumor effect on HCC, and the metabolomics method based on UPLC-Q-TOF/MS combined with further verification at the biochemical level is a promising way to reveal its underlying mechanism.