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. 2026 Feb 20;16:9966. doi: 10.1038/s41598-026-41087-0

Lycium barbarum polysaccharide inhibits Hcy-induced vascular smooth muscle cells migration and invasion via upregulation of KLF4

Xinpeng Ma 1,#, Xiuyu Wang 1,#, Tingrun Mo 1, Xing Ma 1, Minghao Zhang 1,✉
PMCID: PMC13022277  PMID: 41720914

Abstract

This study examined the potential protective effects of Lycium barbarum polysaccharide (LBP) in opposing homocysteine (Hcy)-triggered vascular smooth muscle cells (VSMCs) migration and invasion. Additionally, we examined how Krüppel-like factor 4 (KLF4) participates in the underlying molecular mechanism. Primary human umbilical vein VSMCs were treated with Hcy (100 µmol/L) and divided into five groups: Control, Hcy (100 µmol/L), Hcy + 400 mg/L LBP, Hcy + 600 mg/L LBP, Hcy + 800 mg/L LBP. Cell migration was assessed by scratch assay to screen the optimal drug intervention concentration. KLF4 expression was analyzed via Western blot. Subsequently, cells were treated with Hcy+KLF4 agonist (APTO-235, AP), Hcy+KLF4 inhibitor (Kenpaullone, Ken), and Hcy + LBP+AP. Cell migration and invasion capacities were assessed via scratch assay and Transwell invasion assay. In comparison to the Control group, Hcy notably promoted VSMC migration and invasion and downregulated KLF4 expression. Both the Hcy + 600 mg/L LBP group and the Hcy + AP group exhibited reduced migration/invasion (P < 0.01 vs. Hcy) and upregulated KLF4. Conversely, Hcy + Ken increased migration/invasion and suppressed KLF4. Notably, Hcy + LBP+AP showed the strongest inhibition of migration/invasion and the highest KLF4 upregulation (P < 0.01 vs. Hcy + AP or Hcy + 600 mg/L LBP). LBP inhibits Hcy-induced VSMCs migration and invasion by upregulating KLF4, with synergistic effects observed upon KLF4 agonist co-treatment.

Supplementary Information

The online version contains supplementary material available at 10.1038/s41598-026-41087-0.

Keywords: Lycium barbarum polysaccharide, KLF4, Vascular smooth muscle cells, Hcy, Atherosclerosis

Subject terms: Cell biology, Medical research

Introduction

Atherosclerosis (AS) constitutes a persistent inflammatory disorder initiated through the complex interaction of diverse elements, including abnormal lipid metabolism and compromised immune function. This pathological process involves intricate molecular mechanisms that progressively damage arterial walls1. Clinical research findings indicate that elevated homocysteine levels (HHcy) constituting a distinct contributor to atherosclerotic conditions (AS) development, Hcy-triggered VSMCs migration and invasion represents one of the pathological mechanisms driving AS pathogenesis, but the specific mechanism by which Hcy induces migration and invasion of VSMCs is unknown2–5. KLF4 is an important transcription factor in the human body, which regulates numerous genes in life activities6. Research has revealed that the expression of KLF4 is down-regulated in ovarian cancer tissues, and the down-regulation of KLF4 expression accelerates the proliferation, invasion and migration of ovarian cancer6. KLF4 overexpression decreases the viability, invasion and migration of papillary thyroid cancer cells7. KLF4 regulates unexplained recurrent spontaneous abortion by influencing the invasion and migration abilities of trophoblast cells8. KLF4 is capable of modulating the development and regeneration of VSMCs, and can govern the phenotypic transition of VSMCs via multiple mechanisms10,11. The above-mentioned research suggests that KLF4 plays a significant role in cell migration and invasion, Whether KLF4 exerts a pivotal role in Hcy-induced migration and invasion of VSMCs remains to be elucidated. LBP has been found to have various effects such as lowering blood glucose and blood lipids, anti-inflammation, enhancing body immunity, and anti-aging, and also improves the oxidative stress state of the aorta and delays AS11,13,14. and inhibits the formation of AS by improving the endothelial function of the human vasculature, and promoting the apoptosis of smooth muscle cells13. However, whether LBP can alleviate Hcy-induced migration and invasion of VSMCs and its mechanism is not clear. In this paper, we explored the protective effect of LBP on Hcy-intervened VSMCs migration and invasion from the perspective of regulating KLF4 protein expression, with the aim of providing experimental bases for exploring the new pharmacological effects of LBP and its molecular mechanism of alleviating Hcy-induced atherosclerosis.

Materials and methods

Cell culture and grouping

Primary cultured VSMCs (Suzhou Beinanachuanglian Biotechnology Co., Ltd.), all cells applied in the experiment were at passages 3–7, Add fetal bovine serum to DEME (China Xavier Biotechnology Co., Ltd. batch no: GA241201023) to make it a cell culture medium containing 10% fetal bovine serum, and allow the cells to grow in the above culture medium. When the fusion degree of VSMCs reached 80%, The VSMCs were randomized into a Control group, a 100 µmol/L Hcy (USA, Sigma) group16,17, and three Hcy-LBP (College of Pharmacy, Ningxia Medical University, content 50%, batch no.:JZ20042204) combination groups Hcy + 400/600/800 mg/L LBP12–14, Hcy+AP18 (5 µmol/L), Hcy+Ken19 (2 µmol/L), and Hcy + 600 mg/L LBP + AP (5 µmol/L) groups were intervened for 48 h. Cell migration and invasion abilities were evaluated using Transwell invasion and scratch wound tests.

Cell scratching experiments

We evenly drew two horizontal reference lines on the back of the 6-well plate with a marker pen, then inoculated VSMC suspension into the plate at 100 µL/well (approximately 5 × 103 cells), and after the cells had grown to 90% to 95% fusion, the cells were scratched off the surface of the cells with the tip of a 1000 µL gun in a direction perpendicular to the horizontal lines drawn by a marker, and then the medium was aspirated, and the cells were added respectively to the medium containing 0, Hcy (100 µmol/L), Hcy (100 µmol/L) +400 mg/L LBP group, 100 µmol/L Hcy + 600 mg/L LBP group, 100µmol/L Hcy + 800 mg/L LBP, 100µmol/L Hcy + 5µmol/L AP, 100 µmol/L Hcy + 2 µmol/L Ken, 100µmol/L Hcy + 600 mg/L LBP + 5µmol/L AP were cultured in the medium. The 6-well plates were removed at 0 h and 48 h, and the area of scratches was observed and photographed under a microscope. Image J software was used to measure the scratch area and calculate the scratch migration rate16,17 (percentage of the scratch area calculated by the formula: (0 h scratch area-remaining area of the scratch after 48 h)/0 h scratch area to compare the difference between groups).

Transwell

To evaluate cell migration capacity, we seeded 4 × 105 cells per well into 8-µm pore Transwell chambers using serum-free medium, and the lower chamber was supplemented with 0 and Hcy (100 µmol/L), Hcy (100 µmol/L)+600 mg/L LBP, Hcy (100 µmol/L) + AP(5 µmol/L), (100 µmol/L) + Ken (2 µmol/L), and (100 µmol/L) + 600 mg/L LBP + AP(5 µmol/L) intervened complete medium. After 48 h of incubation, the chambers were removed, unperforated cells were cleared, polyformaldehyde was added to a new chamber to fix the cells, and the fixed cells were stained with crystal violet, photographed, and analyzed using ImageJ software for cell counting, and the outcomes were quantified17. To evaluate the invasive capacity of the cells, serum-free medium was used to inoculate the cells into Transwell chambers at 4 × 105 cells/well. The chambers were pre-coated with Matrigel matrix gel and incubated in a 37 °C incubator for 3 h. The pore sizes of the chambers were 8 μm, and the lower chambers were supplemented with 0, Hcy (100 µmol/L), and Hcy (100 µmol/L)+600 mg/L LBP, respectively, Hcy (100 µmol/L) + AP(5 µmol/L), Hcy (100 µmol/L) + Ken (5 µmol/L), and Hcy (100 µmol/L) + 600 mg/L LBP + AP(5 µmol/L) after intervention with complete medium. After incubation for 48 h, the chambers were retrieved, and the unperforated cells were removed. Polyformaldehyde was added to a new chamber to fix the cells, and then the fixed cells were stained in a chamber containing crystal violet. The cells were photographed and analyzed using ImageJ software for cytometric quantification of the data. The entire procedure was performed in triplicate, and the data were compiled for analysis.

Western blot analysis

To detect protein level changes in VSMCs, we performed western blot analysis using specific antibodies and extracted proteins using specialized protein extraction kits. After protein quantification, a metal bath at 99 °C for 5 min was added to the up-sampling buffer, and we separated proteins by gel electrophoresis, transmembrane, milk-confinement for 2 h, primary antibody incubation at 4 °C (dilution concentration:1:1000) overnight, washing with TBST for 3 times, and secondary antibody incubation at room temperature (dilution concentration:1:5000) for 2 h16,17. TBST was washed again 3 times, and the chemiluminescent horseradish peroxidase substrate was used for development. Finally, ImageJ software was used for grayscale value statistics.

Statistical analyses

GraphPad Prism 10.0 software was used for the analysis. Data are expressed as mean ± standard deviation (x̅ ± s). The t-test was used for comparisons between two groups, whereas one-way ANOVA was applied for multiple group comparisons. For further multiple comparisons, methods such as Tukey’s Honestly Significant Difference (HSD) or Bonferroni were adopted. Statistical significance was set at p ≤ 0.05.

Results

Screening for the effective dose of LBP in Hcy-triggered VSMC migration

To screen for the optimal dose of LBP for intervention in Hcy-triggered VSMCs migration, we examined the effect of various LBP doses on Hcy-triggered VSMCs migration using scratch wound healing assays. The results showed that compared with the control group, the migration ability of VSMCs in the Hcy group was significantly enhanced, and LBP alleviated the migration of VSMCs caused by Hcy (Fig. 1A,B). In terms of the mechanism, the protein expression of KLF4 in VSMCs of the Hcy group was significantly reduced, and different doses of LBP were able to reverse this effect (Fig. 1C,D). Notably, in the Hcy + 600 mg/L LBP subgroup, KLF4 upregulation reached the most prominent level, accompanied by the most substantial reduction in VSMCs migration capacity; therefore, Hcy + 600 mg/L LBP was chosen for use in subsequent experiments.

Fig. 1.

Fig. 1

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Effect of LBP on Hcy-induced migration of VSMCs. (A) Scratch area of VSMCs under light microscope (scale bar: 100 μm); (B) Quantitative analysis of the migration rate of scratch area in each group (n = 5); (C) Western blot detection of protein KLF4 expression in each group; (D) Quantitative analysis of protein KLF4 expression in each group (n = 3). **P < 0.01.

LBP mitigated the Hcy-triggered augmentation of VSMCs migration and invasion

To gain a deeper understanding of how LBP affects the migration and invasion of VSMCs triggered by Hcy, VSMCs were treated with Hcy and simultaneously exposed to 600 mg/L LBP. The Transwell experiment results showed that compared to the control group, the number of migrating (Fig. 2A) and invasive cells (Fig. 2C) in the Hcy group significantly increased. Simultaneously, 600 mg/L LBP significantly inhibited the migration and invasion (Fig. 2B, D).

Fig. 2.

Fig. 2

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Effect of LBP on Hcy-induced migration and invasion of VSMCs. (A) Transwell experimental cell migration pictures under light microscope (0.5% crystal violet staining × 200, scale bar: 200 μm); (B) Transwell experimental cell migration cell number quantification (n = 5); (C) Transwell experimental cell invasion pictures under light microscope (0.5% crystal violet staining × 200, scale bar: 200 μm); (D) Quantification of cell number of Transwell assay cell invasion (n = 5). **P < 0.01.

Validation of the effects of Ken and AP

To further explore the mechanism of LBP to alleviate the migration and invasion of Hcy-triggered VSMCs, Ken, a KLF4 inhibitor, and AP, an agonist, were administered to intervene in VSMCs, the effect of Ken and AP was confirmed via Western blot. Results showed that KLF4 expression was significantly down-regulated in the Ken group relative to the Control group. Moreover, KLF4 expression was significantly up - regulated in the AP group (Fig. 3A, B). These results suggest that Ken can inhibit the expression of KLF4 in VSMCs, while AP can promote the expression of KLF4 in VSMCs. Therefore, in the subsequent research, we used Ken and AP as the tool drugs that affect the expression of KLF4.

Fig. 3.

Fig. 3

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Effects of Ken and AP on KLF4 expression. (A) Western blot detection of protein KLF4 expression in control and inhibitor and agonist groups; (B) Quantitative analysis of protein KLF4 expression in control and inhibitor and agonist groups (n = 3). *P < 0.05.

LBP attenuates Hcy-triggered VSMCs migration and invasion via upregulating KLF4 expression

To assess how LBP impacts Hcy-triggered VSMCs migration and invasion, scratch wound healing and Transwell invasion assays were utilized. Findings from the scratch wound healing assay revealed that, versus the Control group, Hcy notably boosted VSMCs migration capacity and reduced KLF4 expression; In comparison to the Hcy group, the migration ability of VSMCs cells was decreased in the Hcy + 600 mg/L LBP and Hcy + AP groups, and the migration ability of VSMCs cells was enhanced in the Hcy + Ken group; The migration ability of VSMCs was enhanced in the Hcy + AP and Hcy + 600 mg/L LBP groups, whereas it decreased more significantly in the Hcy + LBP+AP group (Fig. 4A,B). Transwell assay Results demonstrated that the number of migrated VSMCs in the Hcy group was significantly increased and KLF4 was significantly down-regulated compared with Control; The number of migrated VSMCs was reduced in the Hcy + 600 mg/L LBP and Hcy + AP groups when compared with the Hcy group. The number of migrated VSMCs cells was decreased in the Hcy + Ken group, and the number of migrated VSMCs cells was increased in the Hcy + Ken group; the decrease in the number of migrated VSMCs cells was more pronounced in the Hcy + LBP +AP group compared to the Hcy + AP and the Hcy + 600 mg/L LBP groups, as shown in Fig. 4C, D,E, F. Western blot analysis demonstrated that compared with the Control KLF4 was significantly down-regulated in the Hcy group; LF4 expression was notably increased in the Hcy + 600 mg/L LBP and Hcy + AP groups versus the Hcy group, while it was decreased in the Hcy + Ken group; KLF4 was more significantly up-regulated in the Hcy + LBP+AP group than in the Hcy + AP and Hcy + 600 mg/L LBP groups (Fig. 4G, H).

Fig. 4.

Fig. 4

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Effect of LBP on Hcy-induced VSMCs migration and invasion via KLF4. (A) Scratch area of VSMCs under light microscope (scale bar: 100 μm); (B) Migration rate of scratch area in each group (n = 5); (C) Pictures of Transwell experimental cell migration under light microscope (scale bar: 200 μm); (D) Pictures of Transwell experimental cell invasion under light microscope (scale bar: 200 μm); (E) Transwell assay cell migration cell number quantification (n = 5); (F) Transwell assay cell invasion cell number quantification (n = 5); (G) Western blot to detect the expression of protein KLF4 in the control group and each intervention group; (H) Quantitative analysis of protein KLF4 expression in the control group and each intervention group (n = 3). *P < 0.05, **P < 0.01.

Discussion

This study is the first to reveal that low KLF4 protein expression plays a significant role in Hcy-induced migration and invasion of VSMCs. KLF4 may be a key transcription factor in the atherosclerotic process caused by hyperhomocysteinemia. LBP can upregulate the expression of KLF4, thereby alleviating the migration and invasion. This discovery helps to reveal the molecular mechanism by which Hcy induces VSMCs to cause atherosclerosis and provides an experimental basis for the development of the pharmacological effects of LBP.

Derived from methionine (Met) metabolism, Hcy is a non-essential amino acid that engages in a complex cycle of Met and cysteine through transsulfurization18. Elevated Hcy is indicative of metabolic dysfunction, and Hcy can contribute to the formation of atherosclerosis through a variety of mechanisms such as inflammation, endothelial dysfunction, along with VSMC proliferation, motility, and phenotypic switch3,18,19. Among them, VSMCs migrating out of the middle membrane layer, stimulated by Hcy, constitutes a key pathological feature in inducing atherosclerotic disease, breaking through the basement membrane to invade the subendothelial layer, and transforming into foam cells by phagocytosis of oxidized lipids20. It was found that Hcy could interfere with the expression balance and enzymatic activity of matrix metalloproteinase-2/9 (MMP-2/9) and its specific tissue inhibitory factor-2 (TIMP-2) in VSMCs, and disrupt the extracellular matrix (ECM) through the up-regulation of MMP-2/9 and the down-regulation of TIMP-2, leading to excessive degradation of the main constituents of the basement membrane (e.g., collagen type IV), The physical barrier to VSMC migration is disrupted, thus inducing VSMC migration in rats and disrupting their normal migratory behavior24,25. Hcy also specifically inhibits endothelial nitric oxide synthase (eNOS) activity, reduces the biosynthesis of nitric oxide (NO), and then activates the PI3K/Akt signaling pathway, The physical barrier restricting VSMC migration is disrupted, thereby promoting VSMC migration in rats and altering their migratory patterns26. The enhancement suggests that reducing the migration and invasion ability of VSMCs could be a target for the treatment of HHcy-induced AS disease. However, previous studies have mainly focused on the mechanisms by which Hcy induces VSMC proliferation and phenotypic transformation of VSMCs17. There have been relatively few studies on the migration and invasion of VSMCs, as these processes are the basis of phenotypic transformation. Therefore, exploring the molecular mechanisms by which Hcy induces VSMC migration and invasion is of great significance. In this study, The migration and invasion of VSMCs induced by Hcy were the focus of our research. At the beginning of the study, we firstly screened the up-regulation of KLF4 and the inhibition of Hcy-induced migration of VSMCs by 600 mg/L LBP through the scratch and Western blot experiments, and this result will provide a clinical research dosage reference. Then we further verified the conclusion that LBP could inhibit the migration and invasion of Hcy-triggered VSMCs via Transwell assay, but the detailed mechanism is still unclear.

KLF4 operates as a regulatory protein featuring three unique functional segments: domains responsible for gene silencing, transcriptional stimulation, and nucleic acid recognition. The protein’s architecture comprises specialized regions that mediate suppression of gene expression, enhancement of transcription processes, and specific interaction with genetic material, Research indicates that KLF4 exhibits widespread expression across multiple organ systems, where it serves vital functions in numerous biological mechanisms7–9. This transcription factor significantly contributes to both normal physiological activities and the pathological progression of atherosclerotic conditions23. Research indicates that atherosclerotic developments, including the accumulation of plaques, increased clustering of inflammatory cells at damaged sites, and the release of cytokines, show significant correlation with KLF4 gene activity. The previous group found that Although high-throughput sequencing has implicated KLF4 in the invasion and migration of Hcy-triggered VSMCs, its critical role in VSMCs migration and invasion induced by Hcy remains undefined in the literature10. Therefore, we hypothesized that LBP may reduce the migration and invasion ability of Hcy-triggered VSMCs via up-regulating KLF4 and thereby reducing the Hcy-triggered VSMCs.

LBP, a biologically active compound extracted from the medicinal Lycium barbarum plant native to Ningxia province, has been widely studied for its pharmacological activities. LBP, a water-soluble polysaccharide derived from Lycium barbarum fruit, is recognized as one of the most critical bioactive components in wolfberry due to its diverse biological activities28. LBP boasts a broad spectrum of medicinal and health - promoting benefits, such as antioxidant, immunomodulatory, antitumor, and neuroprotective effects24. Recent research indicates that LBP possesses notable anti-inflammatory properties. These effects are attained by enhancing the biosynthesis of anti - inflammatory cytokines (e.g., IL-10) while concurrently attenuating the expression of pro - inflammatory mediators25. The level of Hcy in mouse blood is significantly reduced by LBP, The proliferation of Hcy-induced VSMCs is significantly decreased, and their phenotypic transformation is inhibited26,27. However, It is not yet clarified whether LBP can mitigate Hcy-induced VSMCs migration and invasion via KLF4 expression regulation, so we intervened the cells with Hcy, Hcy + Ken, Hcy + AP, Hcy + LBP+AP, respectively, and the results showed that the KLF4 decreased most significantly and Enhanced VSMC migration and invasion were observed in the Hcy + Ken group relative to the Hcy group; the migration and invasion of VSMCs was enhanced compared to the Hcy group; Enhanced migration and invasion of VSMCs were observed in the Hcy + Ken group relative to the Hcy group. Hcy+600 mg/L LBP and Hcy + AP groups compared to the Hcy + LBP+AP group, KLF4 up-regulation was more pronounced, and the migration and invasion ability of VSMCs decreased more significantly, this result suggests that LBP-mediated up-regulation of KLF4 protein contributes to the reduction of VSMC migration and invasion, thereby decreasing cell invasive capacity. In terms of mechanism, as a transcription factor, the expression of KLF4 is known to be influenced by the Wnt/β-catenin33,34, NF-Κb35–37 and oxidative stress/NRF238 signaling pathways. Meanwhile, the research found that LBP can regulate transcription-related pathways such as Wnt/β-catenin39,40, NF-κB40,41, and NRF230,42, but the specific mechanism by which LBP upregulates KLF4 has not been confirmed in previous studies. Therefore, the main contribution of this study is to confirm that LBP can up-regulate the protein expression of KLF4, thereby alleviating the migration and invasion of VSMCs induced by Hcy. This discovery will establish new possible target references for the treatment and prevention of AS, and provide experimental data support for the research of LBP related drugs. The detailed mechanism by which LBP upregulates the expression of KLF4 protein will be the focus of our subsequent research.

In summary, LBP can alleviate Hcy-induced migration and invasion of VSMCs andplay a protective role against Hcy-induced AS, and its mechanism is associated with the up-regulation of KLF4 protein expression by LBP.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1 (2.7MB, pdf)

Acknowledgements

We thank the Key Laboratory of Metabolic Cardiovascular Diseases Research of National Health Commission and Ningxia Key Laboratory of Vascular Injury and Repair Research for providing experimental platforms and technical support.

Author contributions

Xinpeng Ma：Design and implementation of experiments, participation in manuscript writing and data analysis, and responsibility for the overall framework construction of the project.Xiuyu Wang：Co-designed experiments with XM, participated in manuscript writing and partial data analysis, with contributions equivalent to Xinpeng Ma. Tingrun MoResponsible for experimental data collection, assistance with partial experimental operations, and organization of original data records.Xing Ma：Independently completed data analysis and statistics, and performed data processing and visualization using software such as GraphPad Prism 10.0.Minghao Zhang：As the corresponding author, designed the experimental protocol and guided the overall research, wrote the main text of the manuscript, and coordinated manuscript submission and academic communication.Xinpeng Ma and Xiuyu Wang contributed equally to this work.Statement: All authors reviewed the manuscript, approved the final version for publication, and are responsible for the accuracy and integrity of the research content.

Funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the Ningxia Natural Science Foundation Project (2024AAC03200), Scientific Research Project of Higher Education Institutions of the Department of Education of Ningxia Hui Autonomous Region (NYG2024116), Open competition mechanism to select the best candidates for key research projects of Ningxia Medical University (XJKF240312), Ningxia University Student Science and Technology Innovation Research Project in 2025 (S202510752020).

Data availability

All data generated or analysed during this study are included in this published article and its supplementary information files.

Declarations

Competing interests

The authors declare no competing interests.

Ethics statement

The study was approved by the Ethics Committee of Ningxia Medical University (2024-N184) and conducted by the Guide for the Care and Use of Laboratory Animals, and the reporting follows the recommendations in the ARRIVE guidelines.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

These authors contributed equally: Xinpeng Ma and Xiuyu Wang.

References

  • 1.Meyer, G. R. Y.D.et al.Programmed death of macrophages in atherosclerosis: mechanisms and therapeutic targets. Nat. Rev. Cardiol.21 (5), 312–325 (2024). [DOI] [PubMed] [Google Scholar]
  • 2.You, Y. R. et al. Epigenetic modulation of Drp1-mediated mitochondrial fission by inhibition of S-adenosylhomocysteine hydrolase promotes vascular senescence and atherosclerosis. Redox Biol.65, 102828 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Habib, S. S. et al. Homocysteine as a predictor and prognostic marker of atherosclerotic cardiovascular disease: a systematic review and meta-analysis. Eur. Rev. Med. Pharmacol. Sci.27 (18), 8598–8608 (2023). [DOI] [PubMed] [Google Scholar]
  • 4.Huynh, D. T. N. & Heo, K. S.Role of mitochondrial dynamics and mitophagy of vascular smooth muscle cell proliferation and migration in progression of atherosclerosis. Arch. Pharm. Res.44 (12), 1051–1061 (2021). [DOI] [PubMed] [Google Scholar]
  • 5.Sun, F. X., Zeng, Y. & Zhang, M. H. Changes in CTRP9 expression during homocysteine induced proliferation, migration, and phenotype transformation of vascular smooth muscle cells. J. Ningxia Med. Univ.43 (11), 1115–1119 (2021). [Google Scholar]
  • 6.He, Z. H., He, J. & Xie, K. P. KLF4 transcription factor in tumorigenesis. Cell. Death Discovery. 9 (1), 118 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Zhang, C. et al. LINC01210 accelerates proliferation, invasion and migration in ovarian cancer through epigenetically downregulating KLF4. Biomed. Pharmacother. 119, 109431 (2019). [DOI] [PubMed] [Google Scholar]
  • 8.Wang, Q. Z. al.KLF4 overexpression decreases the viability, invasion and migration of papillary thyroid cancer cells. Exp. Ther. Med.18 (5), 3493–3501 (2019). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Tan, Y. L.et al.KLF4 regulates trophoblast function and associates with unexplained recurrent spontaneous abortion. J. Transl Med.22 (1), 922 (2024). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Zhang, X. H. et al. KLF4-PFKFB3-driven glycolysis is essential for phenotypic switching of vascular smooth muscle cells. Commun. Biol.5 (1), 1332 (2022). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Ma, X. et al. Screening of miRNAs related to Hcy promoting proliferation and migration of vascular smooth muscle cells. J. Ningxia Med. Univ.46 (01), 7–14 (2024). [Google Scholar]
  • 12.Zhu, S. et al. Lycium Barbarum polysaccharide protects HaCaT cells from PM2.5-induced apoptosis via inhibiting oxidative stress, ER stress and autophagy. Redox Rep.27 (1), 32–44 (2022). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Zhou, B. J. et al. The Effect of Lycium Barbarum Polysaccharide on the Glucose and Lipid Metabolism: A Systematic Review and Meta-Analysis. J. Am. Nutr. Assoc.41 (6), 618–626 (2022). [DOI] [PubMed] [Google Scholar]
  • 14.Zhu, Y. F., Zhao, Q. P. & Jiang, Y. D. Lycium barbarum polysaccharides attenuates high glucose-induced diabetic retinal angiogenesis by rescuing the expression of miR-15a-5p in RF/6A cells. J. Ethnopharmacol.283, 114651–114662 (2022). [DOI] [PubMed] [Google Scholar]
  • 15.Hu, L. et al. Lycium barbarum polysaccharide alleviates atherosclerosis in mice by up regulating the expression of adiponectin. Drug Eval Stud.40 (03), 292–299 (2017). [Google Scholar]
  • 16.Wang, X. Y. et al. Proliferation, migration and phenotypic transformation of VSMC induced via Hcy related to up-expression of WWP2 and p-STAT3. Plos One. 19 (1), e0296359 (2024). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Wang, X. Y. et al. Hypermethylation of the CTRP9 promoter region promotes Hcy induced VSMC lipid deposition and foam cell formation via negatively regulating ER stress. Sci. Rep.13, 19438 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Nakajima, W. et al. Krüppel-like factor 4 and its activator APTO-253 induce NOXA-mediated, p53-independent apoptosis in triple-negative breast cancer cells. Genes12, 539 (2021). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Xue, J. J. et al. Puerarin attenuates myocardial ischemic injury and endoplasmic reticulum stress by upregulating the Mzb1 signal pathway. Front. Pharmacol.15, 1442831 (2024). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Zhang, M. H. et al. MiR-145 alleviates Hcy-induced VSMC proliferation, migration, and phenotypic switch through repression of the PI3K/Akt/mTOR pathway. Histochem. Cell. Biol.153 (5), 357–366 (2020). [DOI] [PubMed] [Google Scholar]
  • 21.González-Lamuño, D. et al. Hyperhomocysteinemia in Adult Patients: A Treatable Metabolic Condition. Nutrients16 (1), 135 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Zhang, S. et al. Homocysteine promotes atherosclerosis through macrophage pyroptosis via endoplasmic reticulum stress and calcium disorder. Mol. Med.29, 73 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Fu, Z. Y. et al. Vascular smooth muscle cell metabolic reprogramming and phenotypic remodeling in atherosclerosis. Cell. Death Discov30, (2025). [DOI] [PMC free article] [PubMed]
  • 24.Wang, X. Y. et al. Reduced C1q/Tumor Necrosis Factor Related Protein9 Expression Promotes Hcy-Induced VSMCs Migraation via Negative Regulating Endoplasmic Reticulum Stress. Am. J. Biomed. Sci.19 (1), 33–38 (2023). [Google Scholar]
  • 25.Meng, L. P. et al. Polyphenols and Polypeptides in Chinese Rice Wine Inhibit Homocysteine-induced Proliferation and Migration of Vascular Smooth Muscle Cells. J. Cardiovasc. Pharmacol.67 (6), 482–490 (2016). [DOI] [PubMed] [Google Scholar]
  • 26.Bao, X. M. & Zheng, H. C. The effect and possible mechanism of homocysteine on the proliferation and migration ability of rat vascular smooth muscle cells. Shandong Med. J.55 (44), 25–27 (2015). [Google Scholar]
  • 27.Yang, C. et al. Role of Kruppel-like factor 4 in atherosclerosis. Clin. Chim. Acta. 512, 135–141 (2021). [DOI] [PubMed] [Google Scholar]
  • 28.Ai, S. F. et al. Research progress on pharmacological mechanisms of Lycium barbarum polysaccharides.Spec. Econ. Anim. Plant.27 (09), 116–118 (2024). [Google Scholar]
  • 29.Tian, X. J. et al. Extraction, Structural Characterization, and Biological Functions of Lycium Barbarum Polysaccharides: A Review. Biomolecules9 (9), 389 (2019). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Li, Z. Y. et al. Lycium barbarum polysaccharide alleviates DSS-induced chronic ulcerative colitis by restoring intestinal barrier function and modulating gut microbiota. Ann. Med.55 (2), 229023 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Qi, G. X. et al. Homocysteine mediates apoptosis of APOE–/– mouse cardiomyocytes through endoplasmic reticulum stress and the intervention effect of Lycium barbarum polysaccharides. J. Ningxia Med. Univ.41 (02), 109–113 (2019). [Google Scholar]
  • 32.Zhang, M. H. et al. Lycium barbarum polysaccharide protects against Homocysteine-induced Vascular smooth muscle cell proliferation and phenotypic transformation via PI3K/Akt pathway. J. Mol. Histol.51 (6), 629–637 (2020). [DOI] [PubMed] [Google Scholar]
  • 33.Zhou, Z. et al. Cerebral cavernous malformations arise from endothelial gain of MEKK3-KLF2/4 signalling. Nature7 (7597), 122–126 (2016). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Yan, Q. et al. Oncofetal proteins and cancer stem cells. Essays Biochem.66 (4), 423–433 (2022). [DOI] [PubMed] [Google Scholar]
  • 35.Liao, X. et al. Krüppel-like factor 4 regulates macrophage polarization. J. Clin. Invest.121 (7), 2736–2749 (2011). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Han, Z. et al. .HOXA1 participates in VSMC-to-macrophage-like cell transformation via regulation of NF-kappaB p65 and KLF4: a potential mechanism of atherosclerosis pathogenesis. Mol. Med.29 (1), 104 (2023). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Niu, N. et al. Targeting Mechanosensitive Transcription Factors in Atherosclerosis. Trends Pharmacol. Sci.40 (4), 253–266 (2019). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Huang, T. et al. Protective effects of KLF4 on blood-brain barrier and oxidative stress after cerebral ischemia-reperfusion in rats through the Nrf2/Trx1 pathway. Cytokine169, 156288 (2023). [DOI] [PubMed] [Google Scholar]
  • 39.Zhao, Q. et al. Lycium Barbarum Polysaccharides Alleviate Hyperglycemia-Aggravated Cerebral Ischemia/Reperfusion Injury by Up‐Regulating Wnt/β‐Catenin Signaling. Adv. Ther.8, e0045 (2025). [Google Scholar]
  • 40.Qi, Y. et al. Effect of Lycium barbarum polysaccharides on cell signal transduction pathways. Biomed. Pharmacother. 147, 112620 (2022). [DOI] [PubMed] [Google Scholar]
  • 41.Gan, F. et al. Lycium barbarum polysaccharides improve CCl4-induced liver fibrosis, inflammatory response and TLRs/NF‐kB signaling pathway expression in wistar rats. Life Sci.192, 205–212 (2018). [DOI] [PubMed] [Google Scholar]
  • 42.Niu, Z. et al. Inhibition of Iron Death by Lycium barbarum Polysaccharides Ameliorates Myocardial Injury in Sepsis: A Pharmacological Mechanism Study Based on the NRF2/HO-1 Pathway. Food Sci. Nutr.13 (9), e70835 (2025). [DOI] [PMC free article] [PubMed] [Google Scholar]

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Supplementary Materials

Supplementary Material 1 (2.7MB, pdf)

Data Availability Statement

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