脊髓损伤
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Figure 6 ｜ In vivo functional and histological evaluations of the therapeutic effect of cycloheximide after SCI.
We successfully constructed a spinal cord contusion animal model at the T8 level by exposing the spinal cord at the T8 segment (Figure 6A). The cycloheximide group showed an improvement in BMS locomotor test scores by 28 dpi, although there was no significant improvement observed with cycloheximide during the first 3 days following SCI (Figure 6B). Furthermore, motor evoked potentials were improved by 28 dpi in the cycloheximide group (Figure 6C). At the 2-week post-SCI follow-up, analysis of tissue samples revealed that the cycloheximide group showed significantly lower levels of inducible nitric oxide synthase (iNOS) and higher levels of arginase 1 (Arg1) compared with the SCI group. The expression levels of the ferroptosis marker genes Gpx4 and xCT (Slc7a11) were upregulated (P < 0.05; Figure 6D). Notably, histological staining using hematoxylin and eosin did not show significant morphological changes in the heart, kidneys, liver, or lungs of mice with SCI after 6 weeks of cycloheximide treatment (Figure 6E). After cycloheximide treatment, Gpx4 showed enhanced red fluorescence, while Acsl4 showed weaker red fluorescence (Figure 6F and G). Additionally, to further ascertain whether cycloheximide mitigated ferroptosis post-SCI, we assessed the levels of ROS, iron accumulation, and lipid peroxidation within the spinal cord tissues. We noted that cycloheximide ameliorated the ROS levels in microglia/macrophages post-SCI (Additional Figure 1A and B), and reduced the total iron content and MDA levels in spinal cord tissues (Additional Figure 1C and D). In conclusion, these findings provide evidence supporting the potential of cycloheximide to reduce ferroptosis, suppress the production of inflammatory factors, and facilitate functional recovery in the context of SCI.