目的 探讨miR-19b修饰的人脐带间充质干细胞（hUC-MSC）外泌体对炎症性肠病大鼠免疫平衡的影响。 方法 鉴定hUC-MSC外泌体；qRT-PCR检测hUC-MSC外泌体中miR-19b表达。大鼠随机分为对照组、炎症性肠病组、hUC-MSC外泌体组、mimic NC+hUC-MSC外泌体组、miR-19b mimic+hUC-MSC外泌体组，每组12只。除对照组外，其他组大鼠均需构建炎症性肠病模型，建模成功后处理，处理一天一次，持续5周。检测大鼠结肠长度、结肠病理及脾脏指数、胸腺指数；流式细胞术检测外周血辅助性T细胞17（Th17）、调节性T细胞（Treg）占比及Th17/Treg；ELISA检测血清白细胞介素（IL）-10、转化生长因子-β1（TGF-β1）、IL-17水平。 结果 与hUC-MSC组、mimic NC+hUC-MSC组比较，miR-19b mimic+hUC-MSC组外泌体中miR-19b表达升高（P<0.05）。与对照组相比，炎症性肠病组结肠水肿，溃疡，细胞排列紊乱，有大量炎性细胞浸润，结肠长度变短，脾脏指数、胸腺指数、外周血Th17占比、Th17/Treg、血清IL-17水平升高，外周血Treg占比、血清IL-10、TGF-β1水平降低（P<0.05）；与炎症性肠病组比较，hUC-MSC外泌体组、mimic NC+hUC-MSC外泌体组、miR-19b mimic+hUC-MSC外泌体组大鼠结肠水肿，溃疡，细胞排列紊乱及炎性细胞浸润现象减轻，结肠长度增加，脾脏指数、胸腺指数、外周血Th17占比、Th17/Treg、血清IL-17水平降低，外周血Treg占比、血清IL-10、TGF-β1水平升高（P<0.05）；与hUC-MSC外泌体组、mimic NC+hUC-MSC外泌体组比较，miR-19b mimic+hUC-MSC外泌体组大鼠结肠水肿，溃疡，细胞排列紊乱及炎性细胞浸润现象有所改善，结肠长度增加，脾脏指数、胸腺指数、外周血Th17占比、Th17/Treg、血清IL-17水平降低，外周血Treg占比、血清IL-10、TGF-β1水平升高（P<0.05）。 结论 miR-19b修饰的hUC-MSC外泌体可促进炎症性肠病大鼠Th17/Treg平衡，改善免疫功能。
Abstract
Objective To investigate the effect of miR-19b-modified human umbilical cord mesenchymal stem cell (hUC-MSC) exosomes on immune balance in inflammatory bowel disease rats. Methods The hUC-MSC exosomes were identified. QRT-PCR was used to detect miR-19b expression in hUC-MSC exosomes. Rats were stochastically assigned into control group, inflammatory bowel disease group, hUC-MSC exosome group, mimic NC+hUC-MSC exosome group, and miR-19b mimic+hUC-MSC exosome group, each with 12. Except for the control group, inflammatory bowel disease models were established in all other groups, and after successful modeling, they were treated, once a day for 5 weeks. The colon length, colon pathology, spleen index, and thymus index were measured. Flow cytometry was used to test the proportions of peripheral blood helper T cells 17 (Th17), regulatory T cells (Treg), and Th17/Treg. ELISA was used to detect serum interleukin (IL)-10, transforming growth factor-β1 (TGF-β1), and IL-17. Results Compared with the hUC-MSC group and the mimic NC+hUC-MSC group, the miR-19b expression in exosomes was higher in the miR-19b mimic+hUC-MSC group (P<0.05). Compared with the control group, the inflammatory bowel disease group had colon edema, ulcers, disordered cell arrangement, abundant inflammatory cell infiltration, and shortened colon length, the spleen index, thymus index, proportion of Th17 and Th17/Treg in peripheral blood, serum IL-17 increased, while the proportion of Treg in peripheral blood, serum IL-10, and TGF-β1 decreased (P<0.05). Compared with the inflammatory bowel disease group, the hUC-MSC exosome group, mimic NC+hUC-MSC exosome group, and miR-19b mimic+hUC-MSC exosome group showed reduced colon edema, ulcers, disordered cell arrangement, and inflammatory cell infiltration in rats, and increased colon length, the spleen index, thymus index, proportion of Th17 and Th17/Treg in peripheral blood, serum IL-17 declined, while the proportion of Treg in peripheral blood, serum IL-10, and TGF-β1 increased (P<0.05). Compared with hUC-MSC exosome group and mimic NC+hUC-MSC exosome group, the miR-19b mimic+hUC-MSC exosome group showed improvement in colon edema, ulceration, disordered cell arrangement, and inflammatory cell infiltration, and an increase in colon length, the spleen index, thymus index, proportion of Th17 and Th17/Treg in peripheral blood, serum IL-17 declined, while the proportion of Treg in peripheral blood, serum IL-10, and TGF-β1 increased (P<0.05). Conclusions The miR-19b-modified hUC-MSC exosomes can promote Th17/Treg balance and improve immune function in inflammatory bowel disease rats.
关键词
miR-19b /
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人脐带间充质干细胞 /
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外泌体 /
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炎症性肠病 /
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辅助性T细胞17 /
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调节性T细胞
Key words
miR-19b /
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Human umbilical cord-mesenchymal stem cells /
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Exosomes /
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Inflammatory bowel disease /
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T helper cell 17 /
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Regulatory T cells
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参考文献
[1] Liu Z, Wu S, Zhang W, et al. Cordycepin mitigates dextran sulfate sodium-induced colitis through improving gut microbiota composition and modulating Th1/Th2 and Th17/Treg balance[J]. Biomed Pharmacother, 2024, 180: 117394. DOI: 10.1016/j.biopha.2024.117394.
[2] Yuan Y, Hu H, Sun Z, et al. Combining metagenomics, network pharmacology and RNA-seq strategies to reveal the therapeutic effects and mechanisms of qingchang wenzhong decoction on inflammatory bowel disease in mice[J]. Drug Des Devel Ther, 2024, 18(1): 4273-4289. DOI: 10.2147/DDDT.S473688.
[3] Zhang S, Zhong R, Tang S,et al. Metabolic regulation of the Th17/Treg balance in inflammatory bowel disease[J]. Pharmacol Res, 2024, 203(1): 107184-107198. DOI: 10.1016/j.phrs.2024.107184.
[4] Wang J, Pei B, Yan J, et al. HucMSC-derived exosomes alleviate the deterioration of colitis via the miR-146a/SUMO1 axis[J]. Mol Pharm, 2022, 19(2): 484-493. DOI: 10.1021/acs.molpharmaceut.1c00450.
[5] Ma H, Jing Y, Zeng J, et al. Human umbilical cord mesenchymal stem cell-derived exosomes ameliorate muscle atrophy via the miR-132-3p/FoxO3 axis[J]. J Orthop Translat, 2024, 49(1): 23-36. DOI: 10.1016/j.jot.2024.08.005.
[6] Cheng X, Zhang X, Su J, et al. MiR-19b downregulates intestinal SOCS3 to reduce intestinal inflammation in Crohn's disease[J]. Sci Rep, 2015, 5(1): 10397-10406. DOI: 10.1038/srep10397.
[7] 周亚妮, 李若楠. 杨梅素调节cAMP/PKA/CREB信号通路对炎症性肠病大鼠免疫功能的影响[J].中国现代应用药学, 2024, 41(11): 1456-1463. DOI: 10.13748/j.cnki.issn1007-7693.20233703.
Zhou YN, Li RN. Effect of myricetin on immune function in rats with inflammatory bowel disease by regulating the cAMP/PKA/CREB signaling pathway[J]. Chinese Journal of Modern Applied Pharmacy, 2024, 41(11): 1456-1463. DOI: 10.13748/j.cnki.issn1007-7693.20233 703.
[8] Hu D, Huang T, Shigeta M, et al. Electrolyzed hydrogen water alleviates abdominal pain through suppression of colonic tissue inflammation in a rat model of inflammatory bowel disease[J]. Nutrients, 2022, 14(21): 4451-4462. DOI: 10.3390/nu14214451.
[9] Xie F, Xiong Q, Li Y, et al. Traditional Chinese medicine regulates Th17/Treg balance in treating inflammatory bowel disease[J]. Evid Based Complement Alternat Med, 2022, 2022(1): 6275136-6275146. DOI: 10.1155/2022/6275136.
[10]Lan H, Qiu W, Wu J, et al. Formononetin reverses Treg/Th17 imbalance in immune-mediated bone marrow failure mice by regulating the PI3K/Akt signaling pathway[J]. Chin Med, 2024, 19(1): 55-65. DOI: 10.1186/s13020-024-00919-9.
[11]Cheng H, Guan X, Chen D, et al. The Th17/Treg cell balance: a gut microbiota-modulated story[J]. Microorganisms, 2019, 7(12): 583-593. DOI: 10.3390/microorganisms7120583.
[12]Chang Y, Zhai L, Peng J, et al. Phytochemicals as regulators of Th17/Treg balance in inflammatory bowel diseases[J]. Biomed Pharmacother,2021, 141(1): 111931-111941. DOI: 10.1016/j.biopha. 2021.111931.
[13]Wang L, Li T, Cao WX, et al. Acupoint application therapy alleviates pain by regulating immune function through inhibiting TLR4/MyD88/NF-κB p65 signaling in a primary dysmenorrhea rat model[J]. Zhen Ci Yan Jiu, 2024, 49(1): 37-46. DOI: 10.13702/j.1000-0607.20230709.
[14]Din MAU, Wan A, Chu Y, et al. Therapeutic role of extracellular vesicles from human umbilical cord mesenchymal stem cells and their wide therapeutic implications in inflammatory bowel disease and other inflammatory disorder[J]. Front Med (Lausanne), 2024, 11(1): 1406547-1406557. DOI: 10.3389/fmed.2024.1406547.
[15]Che J, Wang H, Dong J, et al. Human umbilical cord mesenchymal stem cell-derived exosomes attenuate neuroinflammation and oxidative stress through the NRF2/NF-κB/NLRP3 pathway[J]. CNS Neurosci Ther, 2024, 30(3): e14454. DOI: 10.1111/cns.14454.
[16]Yang S, Liang X, Song J, et al. A novel therapeutic approach for inflammatory bowel disease by exosomes derived from human umbilical cord mesenchymal stem cells to repair intestinal barrier via TSG-6[J]. Stem Cell Res Ther, 2021, 12(1): 315-325. DOI: 10.1186/s13287-021-02404-8.
[17]许新微.人脐带间充质干细胞来源的外泌体调控巨噬细胞m6A修饰缓解炎症性肠病的机制研究[D]. 镇江: 江苏大学, 2023.
Xu XW. Mechanism study on exosomes derived from human umbilical cord mesenchymal stem cells alleviating inflammatory bowel disease by regulating m6A modification of macrophages[D]. Zhenjiang: Jiangsu University, 2023.
[18] 郭冰心. 人脐带间充质干细胞外泌体运输miR-340治疗放射性肺损伤作用机制研究[D].新乡: 新乡医学院, 2020.
Guo BX. Mechanism study on miR-340 delivered by exosomes from human umbilical cord mesenchymal stem cells in the treatment of radiation-induced lung injury[D]. Xinxiang: Xinxiang Medical University, 2020.
[19]Jia Z, Li L, Zhao P, et al. MicroRNA-451 from human umbilical cord-derived mesenchymal stem cell exosomes inhibits alveolar macrophage autophagy via tuberous sclerosis complex 1/mammalian target of rapamycin pathway to attenuate burn-induced acute lung injury in rats[J]. Biomed Environ Sci, 2024, 37(9): 1030-1043. DOI: 10.3967/bes2024.128.
[20]Jiang L, Wang M, Sun R, et al. Methylation of miR-19b-3p promoter exacerbates inflammatory responses in sepsis-induced ALI via targeting KLF7[J]. Cell Biol Int, 2021,45(8): 1666-1675. DOI: 10.1002/cbin. 11601.
[21]Wu L, Zhang X, Luo L, et al. Altered expression of serum miR-106a, miR-19b, miR-17, and PTEN in patients with idiopathic membranous nephropathy[J]. J Clin Lab Anal, 2021, 35(4): e23737. DOI: 10.1002/jcla.23737.
基金
河南省科技攻关项目（242102310081）
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