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Additional file1: Figure S1. Representative immunohistochemical images of the expression of omentin1 in adipose tissues of mice injected with AAV-NC or AAV-omentin1, and the statistical results of staining intensity of omentin1 were presented, scale bar = 250 μm (n = 3). The experiments were performed in triplicate. Results were expressed as mean ± SD. ##P < 0.01, ####P < 0.0001 vs. the sham group; ****P < 0.0001 vs. the model group, one-way ANOVA. Figure S2. Representative images of hearts of mice injected with AAV-omentin1 or AAV-NC were presented, scale bar = 1 cm (n = 6). Figure S3. Representative western blotting images of the expression of A SIRT4 and B SIRT5 in myocardial mitochondria of mice with fat-specific omentin1 overexpression, and the statistical results were presented (n = 4). Results were gained from three independent experiments and were expressed as mean ± SD. ##P < 0.01, ####P < 0.0001 vs. the sham group. A-B one-way ANOVA. Figure S4. The statistical results of immunofluorescence analysis of Figure 5. Pearson’s correlation coefficient represents the degree of colocalization of A Mfn2, B OPA1, C p-Drp1(Ser616), D Parkin and E PINK1 with mitochondria (n = 3). Results were gained from three independent experiments and were expressed as mean ± SD. ##P < 0.01, ####P < 0.0001 vs. the control group; *P < 0.05, **P < 0.01, ***P < 0.001 vs. the OGD group. A-E one-way ANOVA. Figure S5. Representative images of western blotting analysis of the SIRT3 expression in mitochondria of cardiomyocytes treated with omentin1 and 3-TYP and the statistical result was presented (n = 3). Results were gained from three independent experiments and were expressed as mean ± SD. *P < 0.05 vs. the OGD group; $$$P < 0.001 vs. the OGD + omentin1 group. One-way ANOVA. Table S1. The clinical characteristics of heart failure (HF) patients and healthy subjects

Introduction Traditional Chinese medicine (TCM) provides unique advantages for treatment of ischemic stroke, an aging-related vascular disease. Shengmai powder (GRS) is composed of three active components, specifically, ginsenoside Rb1, ruscogenin and schisandrin A, at a ratio of 6:0.75:6. The main objective of this study was to evaluate the effects of GRS on blood–brain barrier (BBB) dysfunction under conditions of middle cerebral artery occlusion/reperfusion (MCAO/R). Methods C57BL/6J mice subjected to MCAO/R were used as a model to assess the protective effects of varying doses of GRS (6.4, 12.8, and 19.2 mg/kg) on BBB dysfunction. Results GRS reduced cerebral infarct volume and degree of brain tissue damage, improved behavioral scores, decreased water content and BBB permeability, and restored cerebral blood flow. Moreover, GRS promoted expression of zona occludens-1 (ZO-1) and claudin-5 while inhibiting matrix metalloproteinase 2/9 (MMP-2/9) expression and myosin light chain (MLC) phosphorylation. In vitro, GRS (1, 10, and 100 ng/mL) enhanced the viability of bEnd.3 cells subjected to oxygen glucose deprivation/reoxygenation (OGD/R) and decreased sodium fluorescein permeability. Conclusion Consistent with in vivo findings, ZO-1 and claudin-5 were significantly upregulated by GRS in bEnd.3 cells under OGD/R and MMP-2/9 levels and MLC phosphorylation reduced through the Rho-associated coil-forming protein kinase (ROCK)/cofilin signaling pathway. Based on the collective findings, we propose that the TCM compound, GRS, plays a protective role against I/R-induced BBB dysfunction.