Figure 6. Piperine relieves inflammation and myelin destruction in EAE model
(A) Representative histological images of spinal cord sections from piperine- or vehicle- treated EAE mice in preventive model. The sections were prepared on day 22 after MOG’s immunization. Hematoxylin and eosin (H&E) staining for assessing of inflammation (left) and luxol fast-blue (LFB) staining for assessing of demyelination (right).
(B) BBB disruption was imaged using BSA-Cy5.5 at Ex/Em=680/720 nm (left). Mice were i.v. injected with BSA-Cy5.5 (50 mg/kg) and subjected to fluorescence imaging at 6 h post-injection. Quantitative results of total fluorescent counts of BSA-Cy5.5 in regions of interest (right). Values are represented as mean ± SEM (n=3). Data are shown as mean ± SEM and Student’s t-test is performed, *P<0.05.
(C) Myelin imaging was performed with a near-infrared dye, 3,3-diethylthiatricarbocyanine iodide (DBT) (left). DBT dye (0.3 mg/kg) were administered to mice by i.v. injections into the tail vein. The fluorescence imaging of the mouse myelin was acquired 5 min post-injection of DBT with Ex/Em=745/800 nm. Quantitative results of fluorescent intensity of DBT in mice (right). Values are represented as mean ± SEM (n=3). Data are shown as mean ± SEM and Student’s t-test is performed, *P<0.05.
(D-E) Flow cytometry analysis of T lymphocytes. Splenocytes were incubated with APC-CD8+ (A) and FITC-CD4+ (B) antibodies for 2 h at 4 °C in the dark, respectively. Flow cytometry analysis was performed on a BD FACS Caliber equipped with Cell Quest software (BD Biosciences, NJ, USA). Quantitative data of CD8+ and CD4+population were presented in right, respectively. Data are shown as mean ± SEM and Student’s t-test is performed, ***P< 0.001.