ID3O elevated BBB score
Spinal cord injury led to a prominent reduction in BBB score relative to the rats in sham group (Figure 1). However, ID30 administration to SCI rats effectively improved BBB score in dose-based manner relative to untreated group. The BBB score was significantly increased on treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg doses of ID30. Treatment with ID30 at 1.0 mg/kg doses increased BBB score in SCI rats to the level of sham group.
Figure 1. Effect of ID30 on BBB score in SCI rats. Treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg ID30 was followed by determination of BBB score. *P<0.05, **P<0.02 vs. SCI group.
ID30 alleviated water accumulation in SCI rat spinal cord tissues
Water collection in tissues of spinal cord was significantly higher in rats with SCI (Figure 2). Accumulation of water in spinal cord tissues was alleviated significantly on treatment of SCI rats with ID30. Although ID30 significantly prevented SCI induced accumulation collection in spinal cord tissues at 0.25, 0.5, and 0.75 mg/kg concentrations but the effect was maximum at 1.0 mg/kg doses.
Figure 2. Effect of ID30 on spinal tissue water accumulation in SCI rats. ID30 treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg doses was followed by measurement of water content in spinal cord tissues. *P<0.05, **P<0.02 vs. SCI group.
ID30 decreased pro-inflammatory factors
In SCI rats the levels of TNF-α as well as other cytokines such as IL-1β and IL-6 were significantly higher in serum (Figure 3). However, ID30 administration significantly down-regulated SCI mediated higher serum levels of TNF-α and cytokines (IL-1β and IL-6). Decrease in serum TNF-α and cytokine (IL-1β and IL-6) levels by ID30 were significant at 0.25, 0.5, and 0.75 mg/kg doses and maximum at 1.0 mg/kg concentration in SCI rats.
Figure 3. Effect of ID30 on inflammatory cytokines in SCI rats. ID30 treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg doses was followed by determination of serum TNF‑α, IL-1β and IL-6 levels. *P<0.05, **P<0.02 vs. SCI group.
Oxidative stress is inhibited by ID30
In rats with SCI the activities of SOD, CAT and GSH were significantly (P<0.05) higher in serum relative sham group (Figure 4). The MDA level in SCI rat serum was significantly lower than sham group. ID30 treatment alleviated SOD, CAT and GSH activities significantly in SCI rats at 0.25, 0.5, 0.75 and 1.0 mg/kg doses. Moreover, the MDA level was significantly promoted in SCI rats on treatment with 0.25, 0.5, 0.75 and 1.0 mg/kg doses of ID30 in dose-based manner.
Figure 4. Effect of ID30 on oxidative stress factors. The ID30 treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg doses was followed by determination of (A) SOD, (B) CAT and (B) GSH activities. (D) The level of MDA was also determined. *P<0.05, **P<0.02 vs. SCI group.
ID30 decreased caspase activities
The levels of cleaved caspase-3 and -9 in tissues of spinal cord were increased significantly by SCI in rats relative to sham group (Figure 5). ID30 treatment at 0.25, 0.5, 0.75 and 1.0 mg/kg doses alleviated SCI mediated increased cleaved caspase-3 and -9 levels in dose-based manner. Cleaved caspase-3 and -9 expression was significantly alleviated by ID30 treatment at 0.25, 0.5 and 0.75 mg/kg doses in SCI rats. Treatment with ID30 at 1.0 mg/kg concentration reduced cleaved caspase-3 and -9 levels close to that of sham group.
Figure 5. Effect of ID30 on caspase cleavage. ID30 treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg doses was followed by ELISA assay for determination of (A) caspase-3 and (B) caspase-9 activities. *P<0.05, **P<0.02 vs. SCI group.
Inhibition of PGE2 expression by ID30
The expression of PGE2 was significantly (P<0.05) promoted by SCI in rats relative to sham group (Figure 6). ID30 treatment at 0.25, 0.5, 0.75 and 1.0 mg/kg doses suppressed SCI mediated elevation of PGE2 expression in dose-based manner. In SCI rats treatment with 1.0 mg/kg doses of ID30 reduced PGE2 expression close to that of the sham group.
Figure 6. Effect of ID30 on PGE2 expression. Treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg ID30 was followed by evaluation of PGE2 expression. *P<0.05, **P<0.02 vs. SCI group.
Elevated PPAR‑γ and suppressed COX‑2 expression by ID30
The SCI caused a significant suppression in PPAR-γ and elevated COX-2 expression in rats (Figure 7). ID30 administration to SCI rats at 0.25, 0.5, 0.75 and 1.0 mg/kg significantly (P<0.05) elevated PPAR-γ expression and suppressed COX‑2 level in dose-based manner. In SCI rats, promotion of PPAR-γ and reduction of COX‑2 level was maximum by ID30 treatment at 1.0 mg/kg.
Figure 7. Effect of ID30 on PPAR‑γ/COX‑2 expression. Treatment of SCI rats with 0.25, 0.5, 0.75 and 1.0 mg/kg ID30 was followed by evaluation of PPAR‑γ/COX‑2 expression by western blotting. *P<0.05, **P<0.02 vs. SCI group.
ID30 promoted Akt and PI3K activation
In rats subjected to SCI the phosphorylation of PI3K and Akt was significantly lower in tissues of spinal cord (Figure 8). However, ID30 administration of SCI rats at 0.25, 0.5, 0.75 and 1.0 mg/kg doses significantly (P<0.05) elevated PI3K and Akt phosphorylation in dose-based manner. The PI3K and Akt phosphorylation in SCI rats was promoted to maximum level on treatment with 1.0 mg/kg doses of ID30.
Figure 8. Effect of ID30 on PI3K/Akt phosphorylation. The ID30 treatment of SCI rats with 0.25, 0.5, 0.75 or 1.0 mg/kg was followed by western blot assay or determination of phosphorylated PI3K and Akt levels. *P<0.05, **P<0.02 vs. SCI group.