番茄红素通过调控血管平滑肌细胞脂质代谢延缓小鼠动脉粥样硬化进展OA

AIM:This study aimed to investigate the effects of lycopene(LYC)on atherosclerosis(AS)and to elucidate its underlying mechanisms through network pharmacology and molecular docking methodologies.The findings were further validated using an in vitro model of oxidized low-density lipoprotein(ox-LDL)-induced foam cell formation in vascular smooth muscle cells(VSMCs)and an in vivo model of AS in ApoE-/-mice.METHODS:Network pharmacology was utilized to identify the primary targets of LYC in AS,and molecular docking was employed to validate the binding affinity between LYC and its candidate targets.The optimal concentrations of LYC were ascertained using the CCK-8 assay,from which two non-cytotoxic concentrations(2.5 and 5 μmol/L)were selected for further experimentation.The VSMCs were randomly divided into normal control(NC)group,model(ox-LDL)group(80 mg/L ox-LDL),ox-LDL+low-dose(2.5 μmol/L)LYC(LYC-L)group,and ox-LDL+high-dose(5 μmol/L)LYC(LYC-H)group.Western blot was conducted to quantify the protein levels of non-receptor tyrosine kinase SRC,phosphorylated SRC(p-SRC),mitogen-activated protein kinase(MAPK),phosphorylated MAPK(p-MAPK),peroxisome proliferator-activated receptor γ(PPARγ),ATP-binding cassette transporter A1/G1(ABCA1/G1),CD36,lectin-like ox-LDL receptor-1(LOX-1),and markers of VSMC pheno-type switching.Additionally,the DiI-ox-LDL uptake assay was employed to assess the lipid uptake capacity of VSMCs,and immunofluorescence staining was utilized to detect the expression of PPARγ.For in vivo studies,fifty ApoE-/-mice,aged 6 weeks,were randomly divided into NC group,model(AS)group,AS+atorvastatin(AT)group,AS+low-dose(10 mg·kg-1·d-1)LYC(LYC-L)group,and AS+high-dose(20 mg·kg-1·d-1)LYC(LYC-H)group(n=10 per group).The mice in NC group received a normal diet,whereas those in the remaining groups were subjected to a high-fat diet for 14 weeks to induce AS,followed by 8 weeks of intragastric administration.After sacrifice,aortic en face and aortic root were stained with oil red O to evaluate plaque area and lipid deposition.Hematoxylin-eosin and Masson staining were applied to assess plaque area and collagen content.Immunofluorescence staining was used to detect CD36 expression in aortic tis-sues,and serum lipid levels were determined using colorimetric assays.Protein expression levels in aortic tissues were al-so analyzed by Western blot.RESULTS:Network pharmacology and molecular docking analyses predicted a strong bind-ing affinity between LYC and SRC,MAPK,and PPARγ.In cellular experiments,ox-LDL stimulation significantly elevat-ed levels of p-SRC/SRC,p-MAPK/MAPK,CD36,and LOX-1,while reduced levels of PPARγ and ABCA1/G1,accompa-nied by increased lipid uptake,and diminished PPARγ fluorescence intensity compared to the NC group.Co-treatment with 2.5 μmol/L LYC did not significantly reverse most of these changes,except for a notable reduction in p-MAPK/MAPK and a marked increase in PPARγ fluorescence intensity.In contrast,5 μmol/L LYC significantly decreased p-SRC/SRC,p-MAPK/MAPK,CD36,and LOX-1,upregulated PPARγ and ABCA1/G1,reduced lipid uptake,and enhanced PPARγ fluorescence intensity(P<0.05).In the in vivo experiments,the ApoE-/-mice in the AS group showed a signifi-cant increase in plaque area and collagen deposition,as well as elevated levels of p-SRC/SRC,p-MAPK/MAPK,CD36,and LOX-1,and decreased levels of PPARγ and ABCA1/G1 compared to the NC group.However,both AT and LYC-H treatment effectively reversed these pathological changes(P<0.05).The LYC-L group showed similar trends,though the differences did not reach statistical significance.CONCLUSION:Lycopene attenuates the progression of atherosclerosis in ApoE-/-mice,potentially through inhibition of SRC and MAPK phosphorylation,which subsequently activates the PPARγ signaling pathway and modulates VSMC lipid metabolism.