Inhibiting leukocyte‐endothelial cell interactions by Chinese medicine Tongxinluo capsule alleviates no‐reflow after arterial recanalization in ischemic stroke

FIGURE 1. TXL alleviated neurological deficits after tMCAO. Neurological deficits in different groups at 6, 24, and 72 h. Data were expressed as means ± SD. N = 3–7 mice each group. *p < 0.05, **p < 0.01.

FIGURE 2. TXL improved no‐reflow after arterial recanalization in ischemic stroke. (A) Representative CBF images by laser speckle perfusion imaging in different groups at 6, 24, and 72 h. Circles indicate regions of interest showing blood‐supplying zones of left middle cerebral artery. Arrows suggest suppression of no‐reflow in ischemic stroke after treatment of TXL. (B) Quantitative analysis of CBF in the respective ROIs compared to contralateral level in the same zones at 6, 24, and 72 h. N = 3–4 mice each group, n = 2–7 measurements each mouse. **p < 0.01.

FIGURE 3. TXL ameliorated leukocyte‐endothelial cell interactions after tMCAO. Two‐photon imaging of leukocytes labeled by Rhodamine 6G in cortex microvessels at 24 h and 72 h. (A) Representative images of leukocyte‐endothelial cell interactions (aggregation and adhesion) directed by the white arrows, and (B) quantitative data of relative number of adherent and aggregated leukocytes per cm2 in cortex venules. N = 3–4 mice each group, n = 1–12 measurements of leukocytes each mouse. **p < 0.01. Scale bar = 50 μm.

FIGURE 4. TXL reduced leukocyte infiltration and protected neurons after tMCAO. HE‐stained sections of ischemic brain at 24 h and 72 h after tMCAO. N = 3 mice each group. Scale bar = 100 μm.

FIGURE 5. TXL reduced the ratio of neutrophils in peripheral blood after tMCAO. Representative flow cytometry dot plots and quantitative analysis for neutrophils (CD45+ly6G+CD11b+) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. *p < 0.05 and **p < 0.01.

FIGURE 6. Effect of TXL on lymphocytes and N/L in peripheral blood after tMCAO. Representative flow cytometry dot plots and quantitative analysis for lymphocytes and N/L among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. *p < 0.05 and **p < 0.01.

FIGURE 7. TXL reduced the ratio of Tregs in peripheral blood after tMCAO. Representative flow cytometry dot plots and quantitative analysis for Tregs (CD4+CD25+CD127−) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. *p < 0.05.

FIGURE 8. TXL reduced the ratio of Ts cells in peripheral blood after tMCAO. Representative flow cytometry dot plots and quantitative analysis for Ts cells (CD3+CD8+) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. **p < 0.01.

FIGURE 9. Effect of TXL on Th cells in peripheral blood after tMCAO. Representative flow cytometry dot plots (A) and quantitative analysis for Th1 (CD3+CD4+IFN‐γ+) (B), Th2 (CD3+CD4+IL‐4+) (C), Th17 cells (CD3+CD4+IL‐17A+) (D), Th1/Th2 (E), Th1/Th17 (F) and Th2/Th17 (G) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. *p < 0.05 and **p < 0.01.

FIGURE 10. Effect of TXL on B cells in peripheral blood after tMCAO. Representative flow cytometry dot plots (A) and quantitative analysis for B cells (CD3−B220+) (B) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. **p < 0.01.

FIGURE 11. Effect of TXL on DCs in peripheral blood after tMCAO. Representative flow cytometry dot plots (A) and quantitative analysis for plasmacytoid DCs (CD11c+B220+) (B) and myeloid DCs (CD11c+ CD11b+) (C) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. **p < 0.01.

FIGURE 12. Effect of TXL on macrophages in peripheral blood after tMCAO. Representative flow cytometry dot plots (A) and quantitative analysis for macrophages (CD11b+F4/80+) (B), M1 macrophages (CD11b+F4/80+CD86+) (C), and M2 macrophages (CD11b+F4/80+CD206+) (D) among different groups at 24 h and 72 h after tMCAO. N = 4–6 mice each group. *p < 0.05 and **p < 0.01.