基础医学与临床 ›› 2016, Vol. 36 ›› Issue (11): 1493-1498.

• 研究论文 • 上一篇    下一篇

Hsp27联合ADSC对脱细胞神经支架修复小鼠坐骨神经缺损后caspase3的影响

王莹,李文媛,岳文江,刘艳翠,张洋,滕诚毅,孙平   

  1. 牡丹江医学院
  • 收稿日期:2015-11-30 修回日期:2016-05-01 出版日期:2016-11-05 发布日期:2016-10-24
  • 通讯作者: 李文媛 E-mail:yuanyuan19800413@163.com
  • 基金资助:
    国家自然科学基金;黑龙江省自然科学基金项目;黑龙江省卫生计生委立项科研课题;牡丹江医学院科学技术研究项目

Effects of HSP27 and ADSC transplantation on expression of caspase3 after acellular nerve allograft in mice

  • Received:2015-11-30 Revised:2016-05-01 Online:2016-11-05 Published:2016-10-24

摘要: 目的 探讨热休克蛋白27(Hsp27)与脂肪源性干细胞(ADSC)联合应用对小鼠脱细胞同种异体神经支架移植后凋亡相关基因caspase3表达及功能恢复的影响。方法 30只成年C57BL/6小鼠随机分为脱细胞神经支架(ANA)组、ADSC组和Hsp27+ADSC组。坐骨神经功能指数(SFI)、电生理和胫前肌湿重比方法检测神经运动功能的恢复,Western bolt检测神经支架内Hsp27和caspase3的蛋白表达。胆碱乙酰转移酶(ChAT)免疫荧光法检测神经移植体中运动轴突表达,并示踪PKH26标记的移植ADSC。结果 与ADSC组比较,Hsp27+ADSC组神经移植体内Hsp27蛋白表达显著增高,caspase3蛋白表达明显降低,ChAT标记运动轴突表达增强,PKH-26标记的ADSC数量显著增高(P<0.05)。与ANA组比较,ADSC组和Hsp27+ADSC组SFI增高、电生理波幅增高、神经传导速度增快、延迟期缩短、胫前肌湿重比率增高,其中Hsp27+ADSC组神经恢复作用显著优于ADSC组(P<0.05)。结论 Hsp27联合ADSC移植对小鼠脱细胞异体神经支架修复坐骨神经缺损后运动轴突再生和功能恢复的作用优于ADSC组,其机制可能与Hsp2抑制caspase3凋亡信号通路,促进移植的ADSC存活有关。

关键词: 热休克蛋白27, 脂肪源性干细胞, 脱细胞异体神经, 运动轴突再生

Abstract: Objective To study the effect of Heat shock protein 27 (Hsp27) and adipose-derived stem cell (ADSC) transplantation on expression of caspase3 and function recovery after acellular nerve allograft repair of the sciatic nerve gap in mice. Methods A total of 30 healthy C57BL/6 mice were randomly divided into ANA group, ADSC group and Hsp27+ADSC group. The sciatic functions index (SFI), electrophysiology and the rate of tibialis anterior muscle wet weight were used to evaluate nerve regeneration and functional recovery. Observe the protein expression of ChAT and the fluorescence signal of ADSC labeled with PKH-26 in the nerve graft by using immunofluorescence. Results Compared with ANA group, the expression of HSP27 protein, the number of ChAT labeled motor axon and PKH-26 labeled ADSC in Hsp27+ADSC group were increased, however, the expression of caspase protein was decreased (P<0.05). Compared with ANA group, SFI, nerve conduction velocity ,wave amplitude, rate of tibialis anterior wet muscle weight were significantly increased in ADSC group and Hsp27+ADSC group (P<0.05), however, incubation period were significantly decreased in two treatment group (P<0.05), moreover, the effect of Hsp27+ADSC group was more powerful than ADSC group (P<0.05). Conclusion The combined Hsp27 and ADSC transplantation treatment promoted motor axon regeneration and function recovery significantly more than ADSC treatments, which may be related with HSP27 inhabited caspase3 apoptotic signaling pathway, and promoted the survival of the transplanted ADSC.

Key words: HSP27, Adipose-derived stem cell, Acellular nerve allograft, Motor axon regeneration

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