摘要
目的探讨脂肪干细胞(adipose-derived stem cells,ADSCs)来源外泌体对周围神经损伤后再生的影响,为周围神经损伤寻找新的治疗方法。方法将36只成年SD大鼠(雌雄不限,体质量220～240 g)随机分成3组,每组12只。A组为正常对照组,B组为坐骨神经挤压损伤组,C组为ADSCs来源外泌体治疗坐骨神经挤压损伤组。A组仅暴露坐骨神经后直接缝合切口,B、C组制备坐骨神经挤压损伤模型;术后次日开始,A、B组于大鼠尾静脉注射PBS液200μL,C组注射含100μg ADSCs来源外泌体的PBS液200μL,每周注射1次,连续12周。注射结束1周后处死大鼠,取损伤处坐骨神经,分别行大体观察、HE染色观察神经束情况、TUNEL检测坐骨神经雪旺细胞(Schwann cells,SCs)凋亡情况,透射电镜观察坐骨神经超微结构和SCs自噬情况。结果大体观察示A组患肢无明显异常,B、C组患肢瘫痪、肌肉萎缩,但C组瘫痪及肌肉萎缩程度轻于B组。HE染色示,A组神经束膜形态规则;B组神经束形态破坏、束膜不规则,有较多无细胞结构和组织碎片;C组神经束膜较完整,明显优于B组。TUNEL检测示,B、C组SCs凋亡细胞数显著多于A组,B组显著多于C组,差异均有统计学意义(P<0.01)。透射电镜观察示,B、C组SCs自噬体较A组明显增加,但C组少于B组。结论 ADSCs来源外泌体对周围神经损伤后再生有一定促进作用,其机制可能与减少SCs凋亡、抑制其自噬、减轻神经瓦勒变性有关。
Objective To investigate the effects of exosomes from adipose-derived stem cells (ADSCs) onperipheral nerve regeneration, and to find a new treatment for peripheral nerve injury. Methods Thirty-six adultSprague Dawley (SD) rats (male or female, weighing 220-240 g) were randomly divided into 3 groups (n=12). Group Awas the control group; group B was sciatic nerve injury group; group C was sciatic nerve injury combined with exosomesfrom ADSCs treatment group. The sciatic nerve was only exposed without injury in group A, and the sciatic nerve crushinjury model was prepared in groups B and C. The SD rats in groups A and B were injected with PBS solution of 200 μLvia tail veins; the SD rats in group C were injected with pure PBS solution of 200 μL containing 100 μg exosomes fromADSCs, once a week and injected for 12 weeks. At 1 week after the end of the injection, the rats were killed and the sciaticnerves were taken at the part of injury. The sciatic nerve fiber bundles were observed by HE staining; the SCs apoptosis ofthe sciatic nerve tissue were detected by TUNEL staining; the ultrastructure and SCs autophagy of the sciatic nerve wereobserved by transmission electron microscope. Results Gross observation showed that there was no obvious abnormalityin the injured limbs of group A, but there were the injured limbs paralysis and muscle atrophy in groups B and C, and thedegree of paralysis and muscle atrophy in group C were lighter than those in group B. HE staining showed that theperineurium of group A was regular; the perineurium of group B was irregular, and there were a lot of cell-free structuresand tissue fragments in group B; the perineurium of group C was more complete, and significantly well than that of groupB. TUNEL staining showed that the SCs apoptosis was significantly increased in groups B and C than in group A, in groupB than in group C (P<0.01). Transmission electron microscope observation showed that the SCs autophagosomes ingroups B and C were significantly increased than those in group A, but the autophagosomes in group C were significantlylower than those in group B. Conclusion The exosomes from ADSCs can promote the peripheral nerve regeneration.The mechanism may be related to reducing SCs apoptosis, inhibiting SCs autophagy, and reducing nerve Walleriandegeneration.
作者
尹刚
刘蔡钺
林耀发
谢铮
侯春林
林浩东
YIN Gang;LIU Caiyue;LIN Yaofa;XlE Zheng;HOU Chunlin;LIN Haodong(Department of Cell Biology,Naval Military Medical University (Second Military Medical University),Shanghai,200433,P.R.China;Department of Orthopaedic Surgery,Changzheng Hospital,Naval Military Medical University (Second Military Medical University),Shanghai,200003,P.R.China;Department of Plastic and Reconstructive Surgery,Shanghai Ninth People's Hospital,School of Medicine,Shanghai jiao Tong University,Shanghai,200011,P.R.China;Department of Plastic Surgery,Changzheng Hospital,Naval Military Medical University (Second Military Medical University),Shanghai,200003,P.R.China)
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2018年第12期1592-1596,共5页
Chinese Journal of Reparative and Reconstructive Surgery
基金
上海市卫生系统优秀学科带头人培养计划(2017BR034)
中国博士后科学基金(2017M623362
2018T111131)~~
关键词
外泌体
脂肪干细胞
周围神经损伤
神经再生
大鼠
Exosome
adipose-derived stem cells
peripheral nerve injury
nerve regeneration
rat
