Guo Bohua,Wang Jianfeng,Zhang Yinan,et al.Therapeutic effect and related mechanisms of bone marrow mesenchymal stem cell-derived exosomes on giant colon model rats[J].Journal of Clinical Pediatric Surgery,,():786-792.[doi:10.3760/cma.j.cn101785-202502059-015]
Therapeutic effect and related mechanisms of bone marrow mesenchymal stem cell-derived exosomes on giant colon model rats
- Keywords:
- Exosomes; Bone Marrow Mesenchymal Stem Cell; Hirschsprung Disease; Rats; Signal Pathway; Models; Animal; Animals; Laboratory
- Abstract:
- Objective To explore the therapeutic effect of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) on the model of megacolon in rats and elucidate its related mechanisms. Methods BMSCs from bone marrow of normal Sprague-Dawley (SD) rats were isolated,cultured and purified in vitro by total bone marrow adhesion.Exosomes were isolated from BMSCs supernatant by ExoQuick reagent method.Thirty SD rats were randomized into model group (megacolon rat model without any intervention),control group (same operation as model group,normal saline used instead of 0.1% BAC solution); intervention group (rat model of megacolon was interfered with by an injection of 100 μg/mL exosomes dilution 1ml into tail vein,once a day for 7 days).Cultivated SH-SY5Y cells were divided into intervention groups (adding 50 ng to conventional culture medium)/μL BMSCs derived exosomes and control group (conventional culture without any intervention).Hematoxylin-eosin (HE) stain was utilized for observing the pathological structural changes of colon; Real-time fluorescent quantitative polymerase chain reaction (PCR) and Western blot were employed for detecting the expressions of NOTCH,GDNF and Sox10 in colonic tissue.Tetramethylazolamide (MTT) and Transwell cell assays were performed for cell proliferation and migration capability. Results BMSCs of rats were isolated and cultured and exosomes derived from BMSCs successfully extracted.HE stain indicated that the pathological changes in colonic tissue of intervention groups were milder than those of model group.The expressions of NOTCH,GDNF and Sox10 mRNA and protein in colonic tissue of model group were significantly lower than those of control group.The expressions of NOTCH,GDNF and Sox10 was significantly higher in colonic tissue of intervention group than those of model group (F=18.120,P<0.001); The levels of NOTCH,GDNF and Sox10 levels of SH-SY5Y cells were significantly higher in intervention group than those in control group (t=3.130,2.420,and 3.340,P<0.05).The proliferation rate of SH-SY5Y cells at various time points and the number of cells passing through artificial basement membrane in intervention group were higher than those in control group (P<0.05). Conclusions BMSCs derived exosomes may improve the pathological changes of colon in rats with Hirschsprung‘s disease.And their effects are achieved through regulating NOTCH/GDNF/Sox10-related signaling pathways.
References:
[1] Tang CSM,Karim A,Zhong YX,et al.Genetics of Hirschsprung’s disease[J].Pediatr Surg Int,2023,39(1):104.DOI:10.1007/s00383-022-05358-x.
[2] Torroglosa A,Villalba-Benito L,Luzón-Toro B,et al.Epigenetic mechanisms in Hirschsprung disease[J].Int J Mol Sci,2019,20(13):3123.DOI: 10.3390/ijms20133123.
[3] Chai M,Su G,Chen W,et al.Effects of bone marrow mesenchymal stem cell-derived exosomes in central nervous system diseases[J].Mol Neurobiol,2024,61(10): 7481-7499.DOI:10.1007/s12035-024-04032-8.
[4] Jafari D,Shajari S,Jafari R,et al.Designer exosomes: a new platform for biotechnology therapeutics[J].Bio Drugs,2020,34(5):567-586.DOI:10.1007/s40259-020-00434-x.
[5] Wei Y,Chang LJ,Zhou XY.Can exosomes derived from bone marrow-derived stem cells help heal intestinal ischemia/reperfusion injury?[J].Dig Dis Sci,2022,67(11): 4971-4973.DOI:10.1007/s10620-022-07552-2.
[6] Hu YQ,Tao RY,Chen L,et al.Exosomes derived from pioglitazone-pretreated MSCs accelerate diabetic wound healing through enhancing angiogenesis[J].J Nanobiotechnology,2021,19(1):150.DOI:10.1186/s12951-021-00894-5.
[7] 余辉,田东浩,许文耀,等.苯扎氯铵制作的大鼠先天性巨结肠模型肠道菌群变化的研究[J].中华小儿外科杂志,2021,42(2):152-158.DOI:10.3760/cma.j.cn421158-20191108-00625-1. Yu H,Tian DH,Xu WY,et al.Changes of colonic microbiome occurring in benzalkonium chloride-treated Hirschsprung’s disease rat model[J].Chin J Pediatr Surg,2021,42(2):152-158.DOI:10.3760/cma.j.cn421158-20191108-00625-1.
[8] Liu GY,Wu Y,Kong FY,et al.BMSCs differentiated into neurons,astrocytes and oligodendrocytes alleviated the inflammation and demyelination of EAE mice models[J].PLoS One,2021,16(5):e0243014.DOI:10.1371/journal.pone.0243014.
[9] Oliva J.Therapeutic properties of mesenchymal stem cell on organ ischemia-reperfusion injury[J].Int J Mol Sci,2019,20(21):5511.DOI:10.3390/ijms20215511.
[10] Ali M,Pham A,Wang XH,et al.Extracellular vesicles for treatment of solid organ ischemia-reperfusion injury[J].Am J Transplant,2020,20(12):3294-3307.DOI:10.1111/ajt.16164.
[11] Daiyue Y,Yang Y,Zhaorong H,et al.Plasma exosomal miR-199a-3p downregulates cell proliferation and migration in Hirschsprung’s disease by targeting mTOR[J].Pediatr Surg Int,2022,39(1):54.DOI:10.1007/s00383-022-05337-2.
[12] Chen Y,Yuan XJ,Li YL,et al.Circulating exosomal microRNA-18a-5p accentuates intestinal inflammation in Hirschsprung-associated enterocolitis by targeting RORA[J].Am J Transl Res,2021,13(5):4182-4196.
[13] Feng Y,Zhou CG,Zhao F,et al.ZEB2 alleviates Hirschsprung’s-associated enterocolitis by promoting the proliferation and differentiation of enteric neural precursor cells via the Notch-1/Jagged-2 pathway[J].Gene,2024,912:148365.DOI: 10.1016/j.gene.2024.148365.
[14] Chng SH,Pachnis V.Enteric nervous system: lessons from neurogenesis for reverse engineering and disease modelling and treatment[J].Curr Opin Pharmacol,2020,50: 100-106.DOI:10.1016/j.coph.2020.02.001.
[15] Diposarosa R,Bustam NA,Sahiratmadja E,et al.Literature review: enteric nervous system development,genetic and epigenetic regulation in the etiology of Hirschsprung’s disease[J].Heliyon,2021,7(6):e07308.DOI:10.1016/j.heliyon.2021.e07308.
[16] Huang TD,Hou YH,Wang X,et al.Direct interaction of Sox10 with cadherin-19 mediates early sacral neural crest cell migration: implications for enteric nervous system development defects[J].Gastroenterology,2022,162(1):179-192.e11.DOI:10.1053/j.gastro.2021.08.029.
[17] Thongpradit S,Jinawath N,Javed A,et al.Novel SOX10 mutations in waardenburg syndrome: functional characterization and genotype-phenotype analysis[J].Front Genet,2020,11:589784.DOI:10.3389/fgene.2020.589784.
Memo
收稿日期:2025-2-28。
基金项目:浙江省卫生健康科技计划(临床研究应用项目2022KY390)
通讯作者:曾祥勇,Email:278409751@qq.com