Treg Immunomodulation Contributes to the Anti-atherosclerotic Effects of Huxin Formula in ApoE-/- Mice

• Published in: Chinese journal of integrative medicine 2024-10, Vol.30 (10), p.896-905
• Main Authors: Ou, Xiao-min, Cai, Jing, Hu, Xiao-yue, Zeng, Qiao-huang, Lan, Tao-hua, Jiang, Wei
• Format: Article
• Language: English
• Subjects: Medicine; Medicine & Public Health; Original Article
• ISSN: 1672-0415; 1993-0402
• DOI: 10.1007/s11655-024-3663-2

Objective To explore the effects of Huxin formula (HXF) in curtailing atherosclerosis and its underlying mechanism. Methods According to random number table method, 24 specific pathogen free male ApoE -/- mice were randomly divided into model group, HXF low-dose (HXF-L) group (8.4 g/kg daily), HXF high-dose (HXF-H) group (16.8 g/kg daily), and pravastatin (8 mg/kg daily) group in Experiment I ( n =6 per group). C57BL/6J mice served as the control group ( n =6). ApoE -/- mice in HXF-L, HXF-H, pravastatin groups were fed a Western diet and administered continuously by gavage for 12 weeks, while C57BL/6J mice in the control group were fed conventional lab mouse chow for 12 weeks. Further, Tregs were depleted by weekly intraperitoneal injection of purified anti-mouse CD25 antibody (PC61, 250 µg per mouse) for 4 weeks in Experiment II ( n =6 per group). Oil Red O and Masson staining were used to evaluate the plaque area and aortic root fibrosis. The CD4 + CD25 + Foxp3 + Treg counts in the lymph nodes and spleen cells were detected using flow cytometric analysis. The transforming growth factor-β1 (TGF-β1), interleukin (IL)-10, and IL-6 serum levels were examined by MILLIPLEX® MAP technology. Quantitative real-time reverse transcription PCR (qRT-PCR) and Western blot were utilized to assess the expression of TGF-β mRNA and protein in the aorta. The expression of CD4 + T lymphocytes, macrophages and smooth muscle cells in the aortic root were detected by immunofluorescence staining. Results HXF reduced plaque area in ApoE -/- mice ( P