Fig. 8

RIPC combined with HKL achieves better efficacy in mice with SCIRI. A Schematic diagram of experimental design. B Representative immunoblot images showing the effect of HKL and combined therapy on the expression of SIRT3 after spinal cord I/R (n = 5/group). C, D Immunofluorescence was used to detect the effect of HKL and combined therapy on the expression of SIRT3 (in green) in neurons (in red) after spinal cord I/R (n = 5/group). Nuclear was stained with DAPI (in blue). Scale bar = 100 μm. E The mice were functionally scored up to 3d postinjury using the BMS after treatment with HKL or combined therapy (n = 5/group). * p < 0.05, HKL treatment group compared to I/R injury group; # p < 0.05, Combination treatment group compared to HKL treatment group. F Representative footprint images from mice on the third day after injury after treatment with HKL or combined therapy and quantification of the stride length and width (n = 5/group). Blue: frontpaw print; red: hindpaw print. G, H MEP analysis was used as an electrophysiological assessment in mice treated with HKL or combined therapy on day 3 and quantification of the peak-to-peak MEP amplitudes (n = 5/group). I Representative images of Nissl bodies in the anterior horn of the spinal cord and quantification of the number of integrated Nissl bodies per section (n = 5/group). Scale bar = 100 μm. J Representative images of TUNEL-positive apoptotic cells (in red) in spinal cord sections on day 3 after I/R injury and quantification of the number of apoptotic cells in each group (n = 5/group). Neuron was stained with NeuN (in green) and Nuclear was stained with DAPI (in blue). Scale bar = 100 μm. K MnSOD, GSH, CAT, and MDA were measured on day 3 after injury to reflect the level of oxidative stress (n = 5/group). Statistical analysis: mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001