安徽医科大学学报2025，Vol.60Issue(11)：2035-2042,8.DOI:10.19405/j.cnki.issn1000-1492.2025.11.007
脱细胞异体神经移植物通过Bcl-2/Cyt-C/Apaf-1信号通路保护SNI大鼠脊髓前角运动神经元的机制
Mechanism of protection of motor neurons in spinal cord anterior horn of SNI rats by acellular nerve allografts via the Bcl-2/Cyt-C/Apaf-1 signalling pathway
摘要
Abstract
Objective To investigate the protective effects and mechanisms of acellular nerve allografts(ANA)on motor neurons in the spinal cord anterior horn of sciatic nerve injury(SNI)rats.Methods SPF grade male SD rats were randomly divided into normal,model,ANA-bridged(bridge group),and autologous nerve transplantation groups(autograft group),with 6 rats in each group.The SNI rat model was established using the right sciatic nerve clamp method for 10 mm.In the bridge group,the ANA was bridged to the two severed ends of the injured sciatic nerve,and in the autograft group,the autologous nerves were flipped head to tail and then bridged to the two se-vered ends.A spectrophotometer was applied to determine the DNA content in normal nerves and ANA.The foot-print test was used to determine the sciatic nerve function index(SFI)of the rats in each group,the wet weight ra-tio of the anterior tibialis muscle was calculated.The morphology and structure of the anterior horn motor neurons of the spinal cord of each group were observed by HE staining.The immunofluorescence and Western blot were used to detect Apaf-1,Caspase-3,Bcl-2,Bax,and Cyt-C proteins expression in the L4-6 segment of the spinal cord.Results The DNA content in the ANA prepared in this study was significantly lower than that in normal nerves(P＜0.05).Compared with the normal group,the SFI and wet weight ratio of the anterior tibialis muscle were re-duced in the model group(P＜0.001);compared with the model group,both SFI and wet weight ratio of the ante-rior tibialis muscle significantly increased in the bridge group and the autografts group(P＜0.05,P＜0.001),and the SFI and wet weight ratio of the anterior tibialis muscle in the autograft group were higher than those in the bridge group(P＜0.001,P＜0.01).The results of HE staining showed that the motor neurons in the anterior horn of the spinal cord of the normal group were structurally intact and had clear cytosolic boundaries;the neurons in the model group were lysed and necrotic,with blurred cytosolic boundaries;the neurons in the bridge group were less lysed and necrotic,but the nuclear translocation phenomenon could still be seen;the neurons in the autograft group were morphologically and structurally intact with clear cytosolic boundaries.Compared with the normal group,the expression of Apaf-1,Caspase-3,Bax and Cyt-C proteins significantly increased in the model group(P＜0.001,P＜0.01,P＜0.01,P＜0.05).Compared with the model group,the expression of Apaf-1,Caspase-3,Bax,and Cyt-C proteins significantly decreased(P＜0.001,P＜0.05,P＜0.05,P＜0.05);but the expres-sion of Bcl-2 protein significantly increased in the bridge group and the autograft group(P＜0.05).The expression of Apaf-1,Caspase-3,Bax and Cyt-C proteins in the autografts group was lower than that in the bridge group(P＜0.001,P＜0.05,P＜0.05,P＜0.05).Conclusion ANA can exert a protective effect on motor neurons in the anterior horn of the spinal cord of SNI rats by improving the morphology and structure of neurons,increasing the ex-pression of Bcl-2 protein,but decreasing the expression of Cyt-C,Bax,Caspase-3,and Apaf-1 proteins in the spi-nal cord.The mechanism of ANA may be related to the Bcl-2/Cyt-C/Apaf-1-mediated mitochondrial apoptosis sig-naling pathway.关键词
脱细胞异体神经移植物/坐骨神经损伤/Bcl-2/Cyt-C/Apaf-1信号通路/脊髓/运动神经元/大鼠Key words
acellular nerve allografts/sciatic nerve injury/Bcl-2/Cyt-C/Apaf-1 signalling pathway/spinal cord/motor neurons/rats分类
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郑梦圆,郝梓彤,朱清华,田壮壮,锅兴达,郑宇荷,李成,付秀美..脱细胞异体神经移植物通过Bcl-2/Cyt-C/Apaf-1信号通路保护SNI大鼠脊髓前角运动神经元的机制[J].安徽医科大学学报,2025,60(11):2035-2042,8.基金项目
河北省自然科学基金(编号:H2021406056) （编号:H2021406056）
河北省高等学校科学技术研究项目(编号:ZD2020178) （编号:ZD2020178）
承德医学院大学生创新创业训练计划项目(编号:2022014、2024074) Natural Science Foundation of Hebei Provincial(No.H2021406056) （编号:2022014、2024074）
Scientific and Techno-logical Research Project for Higher Education Institutions in Hebei Province(No.ZD2020178) （No.ZD2020178）
College Student Innovation and Entrepreneurship Training Project in Chengde Medical University(Nos.2022014,2024074) （Nos.2022014,2024074）
