To Explore the Mechanism of Nuanxinkang on Alleviating Myocardial Fibrosis in Mice with Myocardial Infarction Through Regulation of Myocardial Cell Metabolic Remodeling Based on Macrophage Polarization
Objective To explore the mechanism of Nuanxinkang (NXK,composed of Ginseng Radix et Rhizoma Rubra and Ilex Pubescen) on alleviating myocardial fibrosis in mice with myocardial infarction by orchestrating myocardial cell metabolic remodeling based on macrophage polarization. Methods (1) C57BL/6J male mice were randomly divided into three groups:sham operation group,model group and NXK administration group(1.65 g·kg-1),with 10 rats in each group. The myocardial infarction model of mice was induced by ligating coronary artery. The drug administration group was given NXK once a day for four weeks. Masson staining was used to detect myocardial fibrosis,and quantitative PCR was used to detect the mRNA expression levels of LDHA,HK2,CD36,CPT-1A,PPAR-γ,PGC-1α,Ndufa8,SDHd,Cyc1,Cox4i2,CD86,iNOS and TNF-α in cardiac tissues. The content of inflammatory factor IL-1β in serum was detected by ELISA.(2)Lipopolysaccharide(LPS)was used to induce the polarization of RAW264.7 macrophages into pro-inflammatory subtype. The supernatant fluid of pro-inflammatory macrophages was prepared and subsequently acted on HL-1 cardiomyocytes. RAW264.7 macrophages and HL-1 cardiomyocytes were divided into control serum group(culture medium with 5％ blank serum and 5％ fetal bovine serum),NXK-contained serum group (culture medium with 5％ NXK-contained serum and 5％ fetal bovine serum),LPS treatment group (culture medium with 5％ blank serum,5％ fetal bovine serum and 200 μg·mL-1 LPS)and NXK-contained serum+LPS treatment group (culture medium with 5％ NXK-contained serum,5％ fetal bovine serum and 200 μg·mL-1 LPS). After 24-h intervention in each group,the lipid in the cells was labeled by Oil Red O staining,and the reactive oxygen species (ROS)level of the cells was detected by DCFH-DA fluorescent probe. Results (1)Compared with the model group,collagen deposition area in cardiac tissue of mice was remarkably reduced (P<0.01) and local ventricular wall damage in the infarcted area was alleviated in NXK group. Also,the mRNA expressions of LDHA,HK2,CD36,CPT-1A,PPAR-γ,Ndufa8,SDHd,Cyc1,Cox4i2,CD86,iNOS and TNF-α were significantly decreased (P<0.05,P<0.01,P<0.001). The mRNA expression of PGC-1α was significantly increased (P<0.05). Serum IL-1β level was obviously decreased(P<0.001).(2)Compared with the LPS group,the deposition of lipid droplets in cytoplasm of HL-1 cardiomyocytes in NXK-contained serum+LPS treatment group was remarkably reduced (P<0.05)and ROS level decreased significantly (P<0.001). Conclusion NXK can alleviate myocardial fibrosis and ventricular remodeling after myocardial infarction in mice,and its mechanism may be related to inhibiting the polarization of pro-inflammatory macrophages,alleviating inflammation level,reducing the production of ROS,reducing the level of glycolysis in cardiac tissue,and improving oxidative phosphorylation and fatty acid metabolism.
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