动物造模,大鼠脊髓损伤修复 中国实验动物信息网 2023-06-12 900

　　Objective: To investigate the effects of MST1/2 inhibitors on the expression of Yes related protein YAP, nerve growth related protein 43, glial fibrillary protein GFAP, apoptosis factor Caspase3, and motor function recovery after spinal cord injury in rats.

　　Methods 105 adult female SD rats weighing 180~200g were selected. Thirty five rats were treated with laminectomy without injection of drugs as the Sham surgery group. Seventy rats were divided into normal saline group, MST1/2 inhibitor group, and intraperitoneal injection of normal saline, 1 mg/kg XMU-MP-1. On the 1st, 3rd, 7th, 14th, 21st, and 28th days after surgery, oblique plate experiments and BBB motor function scores were conducted. Then, rats were euthanized to collect tissues around the spinal cord injury area, and immunofluorescence staining and Western blotting were performed to observe the expression changes of YAP, GAP43, Caspase3, and GFAP in the spinal cord tissues of each group of rats.

　　The results of the oblique plate experiment and BBB motor function score showed that from the 7th day after surgery, the scores of the MST1/2 inhibitor group were significantly higher than those of the saline group, and persisted until the 28th day after surgery, with a statistically significant difference (P<0. 05) 05). Immunoblotting results showed that the YAP expression in the MST1/2 inhibitor group was significantly higher than that in the saline group on the 14th day after surgery, with a statistically significant difference (P<0. 05) 05); On the 14th day after surgery, the expression of GFAP and Caspase3 in the MST1/2 inhibitor group was significantly lower than that in the saline group, and GAP43 expression was found, with a statistically significant difference (P<0. 05) 05). Fluorescence staining showed that the MST1/2 inhibitor group had less inflammatory cell infiltration and formed Nervous tissue structural framework, while the saline group had obvious inflammatory cell infiltration and formed a large number of glial scars. The results of immunofluorescence staining showed that YAP was expressed in neurons in the Sham surgery group, YAP was expressed in Glia in the normal saline group and the MST1/2 inhibitor group, and the number of YAP and GAP43 positive cells in the MST1/2 inhibitor group was significantly higher than that in the other two groups. The number of mature and hypertrophic Astrocyte in the MST1/2 inhibitor group was significantly less than that in the normal saline group; The immunofluorescence double labeling results showed that YAP and GFAP were co expressed.

　　Conclusion High dose application of MST1/2 inhibitor after spinal cord injury can significantly promote the expression of YAP, reduce inflammatory injury reaction, inhibit neuronal apoptosis, improve the microenvironment of spinal cord injury area, promote Astrocyte to form the structural framework of primitive Nervous tissue, which is conducive to neurite regeneration and repair, and promote the recovery of motor function.