[1]王敬业,徐诗嘉,张晓,等.补阳还五汤联合骨髓间充质干细胞外泌体治疗大鼠脊髓损伤的机制研究[J].中国中医骨伤科杂志,2025,33(11):1-8.[doi:10.20085/j.cnki.issn1005-0205.251101]
　WANG Jingye,XU Shijia,ZHANG Xiao,et al.Effect of Buyang Huanwu Decoction Combined with Bone Marrow Mesenchymal Stem Cell Exosomes on Spinal Cord Injury in Rats[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2025,33(11):1-8.[doi:10.20085/j.cnki.issn1005-0205.251101]
点击复制

补阳还五汤联合骨髓间充质干细胞外泌体治疗大鼠脊髓损伤的机制研究()

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第33卷
期数:
2025年11期
页码:
1-8
栏目:
实验研究
出版日期:
2025-11-15

文章信息/Info

Title:
Effect of Buyang Huanwu Decoction Combined with Bone Marrow Mesenchymal Stem Cell Exosomes on Spinal Cord Injury in Rats
文章编号:
1005-0205(2025)11-0001-08
作者:
王敬业1; 徐诗嘉1; 张晓2; 段嘉豪2; 范瑜洁3; 高鑫1; 李硕夫2; 杨雷2; 聂颖2△
1湖南中医药大学第一中医临床学院(长沙,410021)
2湖南中医药大学第一附属医院
3湖南省第二人民医院
Author(s):
WANG Jingye1; XU Shijia1; ZHANG Xiao2; DUAN Jiahao2; FAN Yujie3; GAO Xin1; LI Shuofu2; YANG Lei2; NIE Ying2△
1First Clinical College of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410021,China; 2The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410021,China; 3The Second People's Hospital of Hunan Province,Changsha 410007,China.
关键词:
补阳还五汤;  外泌体;  骨髓间充质干细胞;  脊髓损伤;  氧化应激
Keywords:
Buyang Huanwu decoction;  exosomes;  bone marrow mesenchymal stem cells;  spinal cord injury;  oxidative stress
分类号:
R285.5
DOI:
10.20085/j.cnki.issn1005-0205.251101
文献标志码:
A
摘要:
目的:探讨补阳还五汤联合骨髓间充质干细胞外泌体对大鼠脊髓损伤神经功能恢复作用及其机制研究。方法:SD雄性大鼠40只随机分为正常组、模型组、补阳还五汤组(BYHWD组)、骨髓间充质干细胞外泌体组(BMSCs-Exos组)及补阳还五汤+骨髓间充质干细胞外泌体组(BYHWD+BMSCs-Exos组)。脊髓损伤后第7,14,28天进行BBB神经功能评分,取脊髓组织进行苏木精-伊红(HE)染色和尼氏染色; Western Blot法测定脊髓组织核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、突触素(Syn)蛋白表达; 荧光染色检测活性氧(ROS)的表达量。结果:神经功能评分:与正常组比较,模型组、BYHWD组、BMSCs-Exos组、BYHWD+BMSCs-Exos组神经功能评分均不同程度下降,脊髓损伤后第14天和第28天BYHWD+BMSCs-Exos组神经功能评分优于BYHWD组、BMSCs-Exos组及模型组,差异有统计学意义(P<0.05)。HE染色和尼氏染色显示:模型组大鼠神经元细胞数量减少,脊髓损伤区域内大量空洞坏死,BYHWD组、BMSCs-Exos组及BYHWD+BMSCs-Exos组神经元数量和周边空洞坏死有不同程度改善,BYHWD+BMSCs-Exos组改善最为明显。Western Blot法测定Nrf2、HO-1、Syn蛋白表达与正常组比较,模型组Nrf2及HO-1的蛋白表达均明显上调,差异有统计学意义(P<0.05),Syn的蛋白表达明显下调; 与模型组比较,各给药组Nrf2、HO-1及Syn的蛋白表达均明显上调,差异有统计学意义(P<0.05); 与BYHWD组和BMSCs-Exos组比较,BYHWD+BMSCs-Exos组Nrf2、HO-1及Syn的蛋白表达均明显上调,差异有统计学意义(P<0.05)。荧光染色检测活性氧氧化应激损伤显示:BYHWD组、BMSCs-Exos组及BYHWD+BMSCs-Exos组大鼠脊髓组织活性氧荧光强度高于正常组,低于模型组(P<0.05),与BYHWD组和BMSCs-Exos组比较,BYHWD+BMSCs-Exos组活性氧表达显著降低,差异有统计学意义(P<0.05)。结论:补阳还五汤联合骨髓间充质干细胞外泌体能改善脊髓损伤大鼠的神经功能,可能与加速激活Nrf2/HO-1信号通路上调Syn的蛋白表达、抑制活性氧的生成有关。
Abstract:
Objective:To investigate the effect of Buyang Huanwu decoction(BYHWD)combined with bone marrow mesenchymal stem cell exosome(BMSCs-Exos)on the recovery of neurological function of spinal cord injury in rats and its mechanism.Methods:40 SD male rats were randomly divided into normal group,model group,BYHWD group,BMSCs-Exos group,BYHWD+BMSCs-Exos group.BBB neurological function scores were performed at 7,14,and 28 d after spinal cord injury,and spinal cord tissues were taken for Hematoxylin-eosin(HE)staining and Nissl staining.Nrf2,HO-1,and Syn protein expression were determined by Western Blot with fluorescence staining for reactive oxygen species(ROS).Nrf2,HO-1,and Syn protein expression were determined by Western Blot.Fluorescence staining was used to detect the expression of ROS.Results:Neurological function scores:compared with the normal group,the neurological function scores of the model group,BYHWD group,BMSCs-Exos group,and BYHWD+BMSCs-Exos group decreased to different degrees,and the neurological function scores of the BYHWD+BMSCs-Exos group were better than those of the BYHWD group,the BMSCs-Exos group,and the model group after 14 d and 28 d after the operation(P<0.05).HE staining and Nissl staining showed that:the number of neuronal cells decreased in the model group,and a large number of cavities were necrotic in the area of spinal cord injury.The number of neurons and peripheral cavities necrotic were improved to different degrees in the BYHWD group,the BMSCs-Exos group,the BYHWD+BMSCs-Exos group,and most obviously in the BYHWD+BMSCs-Exos group.The improvement was most obvious in the BYHWD+BMSCs-Exos group.The protein expression of Nrf2,HO-1,and Syn was significantly up-regulated in the model group compared with that in the normal group by Western Blot(P<0.05),and down-regulated in the model group,and significantly up-regulated in each of the drug-delivered groups compared with that in the model group(P<0.05).The protein expression of Nrf2,HO-1,and Syn was significantly up-regulated in the BYHWD+BMSCs-Exos group compared with the BYHWD and BMSCs-Exos groups(P<0.05).Fluorescence staining to detect the oxidative stress damage of ROS showed that the spinal cord tissues of rats in the BYHWD group,the BMSCs-Exos group,the BYHWD+BMSCs-Exos group,and the BYHWD+BMSCs-Exos group were significantly down-regulated(P<0.05).BMSCs-Exos group was higher than the normal group and lower than the model group(P<0.05),and the expression of ROS in BYHWD+BMSCs-Exos group was significantly lower than that in BYHWD group and BMSCs-Exos group(P<0.05).Conclusion:Buyang Huanwu decoction combined with bone marrow mesenchymal stem cell exosomes can improve the neurological function of rats with spinal cord injury,which may be related to the activation of Nrf2/HO-1 signalling pathway to up-regulate the protein expression of Syn and inhibit the generation of ROS.

参考文献/References:

[1] ONG B,WILSON J R,HENZEL M K.Management of the patient with chronic spinal cord injury[J].The Medical Clinics of North America,2020,104(2):263-278.
[2] 韩佃晖,牟青春,陈晓丰,等.创伤性脊髓损伤治疗方式的研究进展[J].中华神经外科疾病研究杂志,2024,18(6):81-84.
[3] 邓建雄,李亮,张亦辉,等.补阳还五汤治疗脊髓损伤的研究进展[J].中国当代医药,2024,31(26):188-193.
[4] CHAI M,SU G,CHEN W,et al.Effects of bone marrow mesenchymal stem cell-derived exosomes in central nervous system diseases[J].Molecular Neurobiology,2024,61(10):7481-7499.
[5] 江朔轩,邓强,张彦军,等.间充质干细胞源性外泌体治疗脊髓损伤的研究进展[J].神经损伤与功能重建,2024,19(5):282-284.
[6] CATANEO A H D,TOMIOTTO-PELLISSIER F,MIRANDA-SAPLA M M,et al.Quercetin promotes antipromastigote effect by increasing the ROS production and anti-amastigote by upregulating Nrf2/HO-1 expression,affecting iron availability[J].Biomedicine & Pharmacotherapy,2019,113:108745.
[7] 龙智生,龚飞鹏,温家宾,等.骨髓间充质干细胞外泌体联合茶多酚治疗大鼠脊髓缺血再灌注损伤[J].中国组织工程研究,2024,28(19):2953-2959.
[8] HU X,XU W,REN Y L,et al.Spinal cord injury:molecular mechanisms and therapeutic interventions[J].Signal Transduction and Targeted Therapy,2023,8:245.
[9] 陈漳鑫,余铃,郭卫春.2024国际脊柱研究学会/Praxis脊髓研究所急性脊髓损伤管理临床实践指南解读[J].临床外科杂志,2025,33(1):52-56.
[10] HWANG B Y,MAMPRE D,AHMED A K,et al.Ultrasound in traumatic spinal cord injury:a wide-open field[J].Neurosurgery,2021,89(3):372-382.
[11] TANG R Z,BOTCHWAY B O A,MENG Y F,et al.The inhibition of inflammatory signaling pathway by secretory leukocyte protease inhibitor can improve spinal cord injury[J].Cellular and Molecular Neurobiology,2020,40(7):1067-1073.
[12] VAN S,BAREYRE F.Chemogenetic approaches to unravel circuit wiring and related behavior after spinal cord injury[J].Experimental Neurology,2021,345:113839.
[13] O'ROURKE S A,SHANLEY L C,DUNNE A.The Nrf2-HO-1 system and inflammaging[J].Frontiers in Immunology,2024,15:1457010.
[14] DODSON M,DE LA VEGA M R,CHOLANIANS A B,et al.Modulating NRF2 in disease:timing is everything[J].Annual Review of Pharmacology and Toxicology,2019,59:555-575.
[15] HE F,RU X L,WEN T.NRF2,a transcription factor for stress response and beyond[J].International Journal of Molecular Sciences,2020,21(13):4777.
[16] 陶经纬,周婧雅,刘奕扬,等.川芎嗪对脊髓损伤大鼠Nrf2/HO-1通路及氧化应激的调控作用[J].中华中医药杂志,2024,39(5):2575-2581.
[17] ZHAO W Y,GASTERICH N,CLARNER T,et al.Astrocytic Nrf2 expression protects spinal cord from oxidative stress following spinal cord injury in a male mouse model[J].Journal of Neuroinflammation,2022,19(1):134.
[18] 吴兵,生梦飞,张想旺,等.木犀草素对脊髓损伤大鼠Nrf2/HO-1通路的影响[J].中国药师,2021,24(10):1833-1837.
[19] ZENG H,LIU N,LIU X X,et al.α-Synuclein in traumatic and vascular diseases of the central nervous system[J].Aging,2020,12(21):22313-22334.
[20] GAUGLER M N,GENC O,BOBELA W,et al.Nigrostriatal overabundance of α-synuclein leads to decreased vesicle density and deficits in dopamine release that correlate with reduced motor activity[J].Acta Neuropathologica,2012,123(5):653-669.
[21] WANG Y C,FENG G Y,XIA Q J,et al.Knockdown of α-synuclein in cerebral cortex improves neural behavior associated with apoptotic inhibition and neurotrophin expression in spinal cord transected rats[J].Apoptosis,2016,21(4):404-420.
[22] 钟占琼,薛清彩,刘晓芬,等.α-突触核蛋白在大鼠脊髓损伤后的表达及意义[J].中国医药导刊,2019,21(3):170-176.
[23] 侯智颖,陈明良.补阳还五汤治疗中医骨伤科疾病研究进展[J].河南中医,2023,43(3):468-474.
[24] 任思思,张赞,李娟,等.补阳还五汤治疗脑病的系统评价和Meta分析[J].时珍国医国药,2021,32(3):745-750.
[25] 李旭,杨阳.补阳还五汤对脊髓损伤大鼠脊髓功能修复及gp130、IL-6的影响[J].时珍国医国药,2023,34(10):2350-2353.
[26] 高鹏宇,潘汉升,王火福,等.补阳还五汤治疗骨伤科疾病及药理研究进展[J].光明中医,2023,38(4):782-785.
[27] 张维平,常雯茜,朱换平,等.加味补阳还五汤联合电针治疗创伤性脊柱脊髓损伤气虚血瘀证的临床疗效[J].中国实验方剂学杂志,2021,27(18):88-93.
[28] 许卢春,姜国正,马昱堃,等.基于GPX4-ACSL4轴探讨补阳还五汤抑制铁死亡促进脊髓损伤后神经功能恢复的作用机制[J].中国实验方剂学杂志,2025,31(5):20-30.
[29] 郭德华,吴成林,许洋,等.补阳还五汤对脊髓损伤大鼠脊髓星形胶质细胞增殖的影响[J].时珍国医国药,2023,34(4):835-838.
[30] JOHNSEN K B,GUDBERGSSON J M,DUROUX M,et al.On the use of liposome controls in studies investigating the clinical potential of extracellular vesicle-based drug delivery systems-A commentary[J].Journal of Controlled Release,2018,269:10-14.
[31] GANGADARAN P,RAJENDRAN R L,LEE H W,et al.Extracellular vesicles from mesenchymal stem cells activates VEGF receptors and accelerates recovery of hindlimb ischemia[J].Journal of Controlled Release,2017,264:112-126.
[32] 梁朝鑫,周安远,王伟,等.骨髓间充质干细胞治疗骨科疾病研究进展[J].陕西医学杂志,2022,51(1):121-124.
[33] YE Z D,ZHENG Y K,LI N N,et al.Repair of spinal cord injury by bone marrow mesenchymal stem cell-derived exosomes:a systematic review and meta-analysis based on rat models[J].Frontiers in Molecular Neuroscience,2024,17:1448777.
(收稿日期:2025-06-17)

备注/Memo

备注/Memo:
基金项目:湖南省自然科学基金项目(2023JJ30472)
湖南省教育厅重点项目(22A0261)
湖南省卫生健康委科研项目(B202304078103)
湖南省中医药管理局课题(B2023028)
湖南中医药大学研究生创新课题(2024CX054)
△通信作者 E-mail:gkny222@163.com(聂颖)
更新日期/Last Update: 2025-11-15