Evaluation of the Chetomin effect on histopathological features in a murine acute spinal cord injury model

Abstract

Background: Several research studies have been focused on improving the treatment and prognosis of acute spinal cord injury, as part of this initiative we investigated the use of Chetomin to reduce the inflammatory response in this pathology.

Methods: An experimental, prospective, cross-sectional study was performed using 42 Wistar rats where we analyzed the effect of Chetomin compared to methylprednisolone administered 1 and 8 h after the spinal cord injury in a murine model.

Results: Chetomin administration 8h post-injury decreased IL-6 and VEGF expression; and, and its administration 1h post-injury decreased NF-kB expression.

Conclusions: Chetomin has anti-inflammatory effects in acute spinal cord injury, whether these effects are observable with other proinflammatory markers should be investigated.

Keywords: Chetomin; HIF-1 α; IL-6; Methylprednisolone; Spinal cord injury; TNF-α; VEGF.

Fig. 1

A) Representative histological sections using H&E staining at 40x of the spinal cord which show the different degree of edema in all groups, it can be appreciated a section of the spinal cord from each group. B) Semi quantitative analysis, the results plotted using a Kruskal Wallis Test shows a significant difference between control group with DMSO 1h, CH-DMSO 1h, DMSO 8h and CH-DMSO 8h.

Fig. 2

A) Representative histological sections using H&E staining at 40x of the spinal cord that show the different degree of hemorrhage in all groups, it can be appreciated a section of the spinal cord from each group. B) Semi quantitative analysis, the results plotted using a Kruskal Wallis, p < 0.05. Test shows a significant difference between control group with DMSO 8h and CH-DMSO 8h.

Fig. 3

A) Representative histological sections using H&E staining at 40x of the spinal cord that show the different degree of pyknosis in all groups, it can be appreciated a section of the spinal cord from each group. B) Semi quantitative analysis, the results plotted using Kruskal Wallis, p < 0.05. Test shows no significant difference was observed between groups.

Fig. 4

A) Representative histological sections using H&E staining at 40x of the spinal cord that show the different degree of infarct in all groups, it can be appreciated a section of the spinal cord from each group. B) Semi quantitative analysis, the results plotted using a Kruskal Wallis, p < 0.05. Test shows a significant difference between control group with CH-DMSO 1h, CH-DMSO 8h, and group CH-DMSO 1h with CH-DMSO 8h.

Fig. 5

A) Representative histological sections using H&E staining at 40x of the spinal cord which show the different degree of polymorphonuclear infiltration in all groups, it can be appreciated a section of the spinal cord from each group. B) Semi quantitative analysis, the results plotted using a Kruskal Wallis, p < 0.05. Test shows a significant difference between control group with CH-DMSO 8h, and MP 8h, also DMSO 1h with MP 8h.

Fig. 6

Bar represents the means and their standard errors for each study group using an ANOVA test post hoc with p < 0.05. Considering all the variables of the histological damage of the study this this includes edema, hemorrhage, pyknosis, infarction, and polymorphonuclear infiltration, no significant difference was observed.

Fig. 7

A) Immunohistochemistry using an antibody of HIF-1α in the spinal cord at 40x. It can be appreciated a section of the spinal cord from each group. B) The results plotted using a Kruskal Wallis Test shows no significant difference between the groups, p < 0.05.

Fig. 8

A) Immunohistochemistry using an antibody of VEGF in the spinal cord at 40x, it can be appreciated a section of the spinal cord from each group. B) The results plotted using Kruskal Wallis, p < 0.05. Test shows a significant difference be-tween control group with CH-DMSO 1h, DMSO 8h, CH-DMSO 8h h and MP 8 h groups; between the DMSO 1 h group and the CH-DMSO 1 h, DMSO 8 h, CH-DMSO 8 h and Mp 8 h groups; and finally, between the DMSO 1 h group and the CH-DMSO 1 h group, where the administration of DMSO and Chetomin with DMSO 8 h after injury decreased the VEGF level.

Fig. 9

A) Immunohistochemistry using an antibody against IL-6 in the spinal cord at 40 × ; a section of the spinal cord from each group is shown. B) The results were plotted using the Kruskal–Wallis test, p < 0.05. B) Test showing a significant difference between the control group and the DMSO 1 h, CH-DMSO 1 h, CH-DMSO 8 h and MP 8 h groups and between the DMSO 1 h group and the DMSO 8 h group, where the administration of Chetomin 8 h after injury decreased the level of IL-6.

Fig. 10

A), Immunohistochemistry was performed using an antibody against NF-κB in the spinal cord at 40 × , and a section of the spinal cord from each group was obtained. B), The results were plotted using the Kruskal Wallis test, p < 0.05. The test revealed a significant difference between the control group and the DMSO 1 h, CH-DMSO 1 h, and DMSO 8 h groups, where the administration of Chetomin 1 h after injury decreased the level of NF-κB.

Fig. 11

Immunohistochemistry using an antibody against 3-NT in the spinal cord at 40 × revealed a section of the spinal cord from each group. The results plotted using a Kruskal–Wallis test show a significant difference between DMSO for 1 h and CH-DMSO for 8 h and between MP for 8 h and CH-DMSO for 1 h and CH-DMSO for 8 h and MP for 8 h.