骨髓间充质干细胞外泌体对神经病理性大鼠镇痛、病理及anx1-Src-NMDAR-2B的影响

Effects of BMSCs exosomes on analgesic effect,pathological morphology and Panx1-Src-NMDAR-2B in neu ropathic rats

目的 探讨骨髓间充质干细胞(BMSCs)外泌体对神经病理性大鼠镇痛作用、病理形态及Panx1-Src-NMDAR-2B的影响。方法 30只SD雄性大鼠,随机分为正常(NO)组,模型(MO)组,BMSCs外泌体(BE)组,各10只,对MO组、BE组采用坐骨神经慢性压迫性损伤建立神经病理性大鼠模型,NO组不建立该模型,建模成功后,对BE组鞘内注射0.5 mL 200 mg/L的BMSCs外泌体,NO组、MO组同期鞘内注射同体积生理盐水,用自发疼痛行为学评分、热痛阈值、机械疼痛阈值评价镇痛作用,苏木精-伊红(HE)染色检测脊髓组织病理形态,免疫印迹法检测Panx1-Src-NMDAR-2B蛋白表达。结果 NO组、MO组、BE组建模成功后、给药1周末、给药2周末自发疼痛行为学评分、热痛阈值、机械痛阈值差异均有统计学意义(P<0.05);NO组脊髓组织形态正常,MO组出现明显变化,与MO组相比,BE组病理形态明显改善;NO组、MO组、BE组脊髓组织Panx1蛋白表达分别为0.96±0.06、2.36±0.21、1.54±0.16(F=20.270,P<0.01),Src蛋白表达分别为1.24±0.12、2.29±0.24、1.61±0.19(F=12.370,P<0.001),NMDAR-2B蛋白表达分别为1.25±0.11、2.32±0.26、1.59±0.18(F=11.990,P<0.001),NO组、MO组、BE组脊髓组织Panx1、Src、NMDAR-2B蛋白表达异均有统计学意义(P<0.05)。结论 BMSCs外泌体对神经病理性大鼠具有显著疗效,具有显著的镇痛作用,并可有效改善大鼠病理形态,抑制Panx1-Src-NMDAR-2B通路表达。

ObjectiveeTo investigate the effects of BMSCs exosomes on analgesic effect,pathological morphology and Panx1-Src-NMDAR-2B in neuropathic rats.Methods Thirty SPF grade SD male rats were randomly divided into normal(NO)group,model(MO)group and BMSCs exosome(BE)group,with 10 rats in each group.Chronic compression injury of sciatic nerve was used to establish neuropathic rat models in MO and BE groups,while the model was not established in NO group.In the BE group,O.5 mL 200 mg/L BMSCs exosomes were injected intratherally,and in the NO and MO groups,the same volume of normal saline was injected intratherally at the same time.Analgesic effects were evaluated by spontaneous pain behavioral scores,thermal pain threshold and mechanical pain threshold,and pathological morphology of spinal cord was detected by HE.The expression of Panx1-Src-NMDAR-2B protein was detected by western blotting.Results The behavioral scores of spontaneous pain,thermal pain threshold and mechanical pain threshold of NO group,MO group and BE group were statistically significant after the successful formation of the model,one weekend of administration and two weeks of administration(P<0.05).The spinal cord morphology of NO group was normal,and the pathological morphology of MO group was significantly improved compared with MO group.The expression of Panx1 protein in the spinal cord of NO group,MO group and BE group was 0.96±0.06,2.36±0.21 and 1.54±0.16,respectively(F=20.270,P<0.001).Src protein expression was 1.24±0.12,2.29±0.24,1.61±0.19(F=12.370,P<0.001),and NMDAR-2B protein expression was 1.25±0.11,2.32±0.26,1.59+0.18(F=11.990,P<0.001),the protein expressions of Panx1,Src and NMDAR-2B in the spinal cord of NO,MO and BE groups were statistically significant(P<0.05).Conclusion BMSCs exosomes have significant curative effect on neuropathic rats,have significant analgesic effect,and can effectively improve the pathological morphology of rats,inhibit the expression of Panx1-Src-NMDAR-2B pathway.