脂肪干细胞移植治疗1-溴丙烷所致大鼠周围神经损伤

Adipose-derived stem cell transplantation for the treatment of peripheral nerve injury induced by 1-bromopropane in rats

目的探讨脂肪干细胞(ADSCs)移植对1-溴丙烷致大鼠周围神经损伤的修复作用。方法选择无特定病原体级SD雄性大鼠随机分为对照组(15只)和染毒组(30只)。染毒组大鼠予剂量为800 mg/kg体质量的1-溴丙烷灌胃染毒,对照组予等体积玉米油灌胃,每日1次,每周5 d,连续染毒8周。建立1-溴丙烷所致大鼠周围神经损伤模型后,随机将染毒组大鼠分为自行恢复组和ADSCs组,每组12只。ADSCs组大鼠予尾静脉注射0.5 mL(含1×10^(6)个)ADSCs细胞悬液,自行恢复组大鼠予等体积磷酸盐缓冲液,每周1次,连续4周;对照组大鼠不予任何处理。在ADSCs治疗前1天和治疗后第7、14、21、28天,对各组大鼠进行斜板实验和BBB运动功能评分。治疗结束后,分离大鼠坐骨神经进行组织病理学检查,采用比色分析法测定大鼠大脑运动皮质中氧化应激指标,采用酶联免疫吸附测定法检测大鼠坐骨神经中脑源性神经营养因子(BDNF)和神经生长因子(NGF)的水平。结果自行恢复组大鼠治疗的5个时间点斜板实验最大倾斜角度和BBB评分均低于对照组(P值均<0.05);与自行恢复组比较,ADSCs组大鼠治疗第21和28天斜板实验最大倾斜角度均增大(P值均<0.05),治疗第7、14、21和28天BBB评分均增加(P值均<0.05)。与对照组比较,自行恢复组大鼠大脑运动皮质中丙二醛水平增加(P<0.05),超氧化物歧化酶(SOD)活力和谷胱甘肽水平均降低(P值均<0.05),坐骨神经中BDNF和NGF水平均降低(P值均<0.05)。ADSCs组大鼠大脑运动皮质中SOD活力和丙二醛、谷胱甘肽水平,以及坐骨神经中BDNF和NGF水平分别与对照组比较,差异均无统计学意义(P值均>0.05)。与自行恢复组比较,ADSCs组大鼠大脑运动皮质中SOD活力和谷胱甘肽水平均增加(P值均<0.05),丙二醛水平降低(P<0.05),坐骨神经中BDNF表达水平增加(P<0.05)。结论ADSCs移植可通过抗氧化应激和调节神经营养因子的分泌而改善1-溴丙烷致大鼠周围神经功能损伤。

Methods To investigate the repairing effect of adipose-derived stem cells(ADSCs)transplantation in 1-bromopropane-induced peripheral nerve injury in rats.Methods A total of 45 specific pathogen free male SD rats were randomly assigned to the control group(15 rats)and exposure group(30 rats).The rats in the exposure group were gavaged with 1-bromopropane at a dose of 800 mg/kg body weight,while the control group were given with equal volume of corn oil,once a day,five days per week for eight weeks.The rat model of peripheral nerve injury induced by 1-bromopropane was established and was randomly assigned to the self-recovery group and ADSCs group,12 rats in each group.ADSCs group and self-recovery group were injected with 0.5 mL(contains 1×10^(6)cells)ADSCs or 0.5 mL phosphate buffer solution respectively by tail vein,once a week for four weeks.The control group was not administered any treatment.The rats in each group were assessed using inclined plane test and Basso-Beattie-Bresnahan(BBB)score on the first day before treatment as well as 7,14,21 and 28 day after treatment.At the end of treatment,the sciatic nerve was isolated for histopathological examination.The oxidative stress indexes in the cerebral motor cortex were detected by colorimetric analysis.The levels of brain-derived neurotrophic factor(BDNF)and nerve growth factor(NGF)in the sciatic nerve were detected by enzyme-linked immunosorbent assay.Results The maximum tilt angle of the inclined plate and the BBB score were lower in self-recovery group at the five time points of treatment(all P<0.05)compared with the control group.The maximum tilt angle of the inclined plate test was higher in ADSCs group at 21and 28 days of treatment(all P<0.05),and the BBB score was higher at 7,14,21 and 28 days of treatment(all P<0.05),when compared with the self-recovery group.The level of malondialdehyde(MDA)in the cerebral motor cortex increased(P<0.05),while the superoxide dismutase(SOD)activity and glutathione(GSH)level decreased(all P<0.05),and the levels of BDNF and NGF in the sciatic nerve decreased(all P<0.05)in self-recovery group compared with the control group.There was no significant difference in SOD activity and MDA,GSH,BDNF and NGF levels between ADSCs group and control group(all P>0.05).The level of MDA in cerebral motor cortex decreased(P<0.05),the SOD activity and GSH level increased(all P<0.05),and the level of BDNF in sciatic nerve increased(P<0.05)in ADSCs group compared with the self-recovery group.Conclusion ADSCs transplantation can repair peripheral nerve injury induced by 1-bromopropane via anti-oxidative stress and regulating the secretion of neurotrophic factors.