Figure 2.
Mn porphyrin administration to TS mice after the inflammatory challenge reverses cell adhesion and leukocyte recruitment, and it restores blood flow by reducing SSRBC ROS production. Anesthetized TS mice were treated subcutaneously with vehicle, 0.1, 0.2, or 2 mg/kg MnBuOE, or 0.5 mg/kg or 2 mg/kg MnE, as per protocol 1. (A) Representative images of postcapillary venules (20× magnification) from vehicle-treated and compound-treated TS mice are presented. Vessels without adherent cells appear gray due to the rapidly moving fluorescence-labeled cells. Sickle RBC and leukocyte adhesion in inflamed vessels and vaso-occlusion are indicated with arrows. Scale bars, 50 μm. (B-D) Video frames of vessel and arteriole segments were used to quantify adhesion of fluorescence-labeled cells (sickle RBCs and leukocytes) presented as the number of adherent cells per 100 μm vessel length and fluorescence unit [FU], and rolling flux of leukocytes. The number of leukocytes (identified based on their size) rolling across a specific point per minute was counted. At least 50 different locations were analyzed for rolling of leukocytes in the vessels, and numbers were averaged among groups of TS mice. (A-C) Before and after treatment with vehicle, TS mice showed marked cell adhesion and vaso-occlusion. Treatment with 0.1, 0.2, or 2 mg/kg MnBuOE or 0.5 or 2 mg/kg MnE reversed cell adhesion and vaso-occlusion. (E) Blood flow in TS mice. Percentage of vessels with normal, slow, and no blood flow in vessels with a venular diameter almost similar (∼25 μm) among all groups tested. (F) The Kaplan-Meier survival curves for MnE-treated (n = 9) or vehicle-treated (n = 11) TS mice. Survival was significantly improved in the MnE-treated group compared with vehicle control. Log-rank (Mantel-Cox) test, P = .0009. (G) ROS generation in sickle RBCs in vivo. Compared with vehicle treatment, TS mice treated with MnBuOE at 0.1, 0.2, and 2 mg/kg and MnE at 0.5 and 2 mg/kg manifest significantly lower SSRBC ROS measurements of DCF-derived signal. (B-D,G) Error bars show standard error of the mean (SEM) of 4 different experiments for each treatment group. *P < .05, **P < .01, ***P < .001, and ****P < .0001 compared with vehicle-treated animals regardless of the vessel diameter.

Mn porphyrin administration to TS mice after the inflammatory challenge reverses cell adhesion and leukocyte recruitment, and it restores blood flow by reducing SSRBC ROS production. Anesthetized TS mice were treated subcutaneously with vehicle, 0.1, 0.2, or 2 mg/kg MnBuOE, or 0.5 mg/kg or 2 mg/kg MnE, as per protocol 1. (A) Representative images of postcapillary venules (20× magnification) from vehicle-treated and compound-treated TS mice are presented. Vessels without adherent cells appear gray due to the rapidly moving fluorescence-labeled cells. Sickle RBC and leukocyte adhesion in inflamed vessels and vaso-occlusion are indicated with arrows. Scale bars, 50 μm. (B-D) Video frames of vessel and arteriole segments were used to quantify adhesion of fluorescence-labeled cells (sickle RBCs and leukocytes) presented as the number of adherent cells per 100 μm vessel length and fluorescence unit [FU], and rolling flux of leukocytes. The number of leukocytes (identified based on their size) rolling across a specific point per minute was counted. At least 50 different locations were analyzed for rolling of leukocytes in the vessels, and numbers were averaged among groups of TS mice. (A-C) Before and after treatment with vehicle, TS mice showed marked cell adhesion and vaso-occlusion. Treatment with 0.1, 0.2, or 2 mg/kg MnBuOE or 0.5 or 2 mg/kg MnE reversed cell adhesion and vaso-occlusion. (E) Blood flow in TS mice. Percentage of vessels with normal, slow, and no blood flow in vessels with a venular diameter almost similar (∼25 μm) among all groups tested. (F) The Kaplan-Meier survival curves for MnE-treated (n = 9) or vehicle-treated (n = 11) TS mice. Survival was significantly improved in the MnE-treated group compared with vehicle control. Log-rank (Mantel-Cox) test, P = .0009. (G) ROS generation in sickle RBCs in vivo. Compared with vehicle treatment, TS mice treated with MnBuOE at 0.1, 0.2, and 2 mg/kg and MnE at 0.5 and 2 mg/kg manifest significantly lower SSRBC ROS measurements of DCF-derived signal. (B-D,G) Error bars show standard error of the mean (SEM) of 4 different experiments for each treatment group. *P < .05, **P < .01, ***P < .001, and ****P < .0001 compared with vehicle-treated animals regardless of the vessel diameter.

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