米托蒽醌甲磺酸盐预处理脂肪间充质干细胞对犬糖尿病的治疗效果评价

doi:10.11843/j.issn.0366-6964.2024.03.043

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (3): 1328-1344.doi: 10.11843/j.issn.0366-6964.2024.03.043

• 临床兽医 • 上一篇

米托蒽醌甲磺酸盐预处理脂肪间充质干细胞对犬糖尿病的治疗效果评价

谭宁¹, 李巴仑¹, 韩苗¹, 李琛琛¹, 景远翔¹, 寇正¹, 李娜¹, 彭莎¹, 赵献军^1,2, 华进联^1*

1. 西北农林科技大学动物医学院陕西省干细胞工程技术研究中心, 杨凌 712100;
2. 百欧派(天津)生物技术有限公司, 天津 300350

收稿日期:2023-06-14 出版日期:2024-03-23 发布日期:2024-03-27
通讯作者: 华进联,主要从事干细胞与生殖发育调控、干细胞抗病育种及重大疾病防控研究,Tel:029-87091117,E-mail:jinlianhua@nwsuaf.edu.cn
作者简介:谭宁(1998-),女,黑龙江宁安人,硕士生,主要从事干细胞治疗与应用研究,E-mail:tanning2020@nwafu.edu.cn;李巴仑(1995-),男,新疆石河子人,博士生,主要从事干细胞治疗与应用研究,E-mail:libalun@nwafu.edu.cn。
基金资助:
国家自然科学基金（32072806）；国家重点研发计划项目（2022YFD1302201）；陕西省重点科技创新团队项目（2019TD-036）；百欧派（天津）生物技术有限公司项目（K4040121281）

Evaluation of Therapeutic Effect of Mitoquionl Mesylate Pretreated Adipose Derived Mesenchymal Stem Cells on Canine Diabetes Mellitus

TAN Ning¹, LI Balun¹, HAN Miao¹, LI Chenchen¹, JING Yuanxiang¹, KOU Zheng¹, LI Na¹, PENG Sha¹, ZHAO Xianjun^1,2, HUA Jinlian^1*

1. College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A & F University, Yangling 712100, China;
2. BaiOuPai (Tianjin) Biotechnology Co., Ltd, Tianjin 300350, China

Received:2023-06-14 Online:2024-03-23 Published:2024-03-27

摘要/Abstract

摘要： 本研究在高脂饮食（high-fat diet，HFD）饲喂和链脲佐菌素（streptozotin，STZ）注射的联合作用下成功建立小鼠及犬糖尿病模型，旨在探究米托蒽醌甲磺酸盐（mitoquionl mesylate，MitoQ）预处理脂肪间充质干细胞（adipose-derived mesenchymal stem cells，ADMSCs）对糖尿病动物模型治疗效果的提升作用及机制。取正常培养状态的ADMSCs及添加1 μmol·L^-1 MitoQ 处理的ADMSCs，对MitoQ处理的细胞形态变化、生长、增殖、迁移及抗氧化能力等进行检测。选取48只8周龄的昆明（KM）雄性小鼠和12只中华田园犬，随机分为正常对照组（NC组）、糖尿病模型组（Diabetes组）、ADMSCs单独治疗组（ADMSCs组）和MitoQ预处理ADMSCs联合治疗组（MitoQ-ADMSCs组）。常规方法制作小鼠及犬糖尿病模型，对模型小鼠和犬分别进行ADMSCs和MitoQ-ADMSCs细胞静脉移植治疗，细胞数为2×10⁶·只^-1（小鼠）和1×10⁷·只^-1（犬），每周1次，连续3周。持续监测临床指标变化，治疗结束后1周收样，血清学、组织学、氧化应激及犬血清代谢组学分析。结果表明，MitoQ预处理ADMSCs不会影响其细胞形态，但能促进其生长、增殖及迁移能力，同时增强其抗氧化能力；ADMSCs和MitoQ-ADMSCs移植治疗后，发现MitoQ显著促进ADMSCs降低血糖的能力，且提高胰岛β细胞功能和机体血糖调节能力；ADMSCs经MitoQ预处理后有效减轻糖尿病伴发的肝代谢功能障碍和血脂代谢异常；ADMSCs可以减轻胰腺和肝组织损伤，促进胰岛素分泌，减轻肝糖原合成障碍和纤维化水平，而MitoQ处理能够显著改善ADMSCs治疗效果；MitoQ预处理ADMSCs能够有效提升机体的抗氧化能力，减轻氧化损伤，进而增强治疗效果；MitoQ预处理ADMSCs可以显著促进机体内与抗氧化相关的代谢物表达上调。结果揭示，MitoQ能够通过提高ADMSCs的抗氧化能力、加速组织损伤修复，从而更好地治疗小鼠及犬糖尿病。

关键词: 米托蒽醌甲磺酸盐, 脂肪间充质干细胞, 犬, 糖尿病

Abstract: In this study, we successfully established mouse and canine diabetes models under the combined effect of high-fat diet (HFD) feeding and streptozotocin (STZ) injecting, aiming to investigate the effect and mechanism of Mitoquionl Mesylate (MitoQ) pretreatment of adipose-derived mesenchymal stem cells (ADMSCs) in enhancing the therapeutic effect of diabetic animal models. At the cellular level, ADMSCs in normal culture were pretreated with MitoQ at a concentration of 1 μmol·L^-1 in α-MEM+ cell culture medium, and the morphological changes, growth, proliferation, migration and antioxidant capacity of MitoQ-ADMSCs and ADMSCs were examined respectively. At the animal level, forty-eight 8-week old Kun-Ming(KM) male mice and 12 Chinese Field canines were selected and randomly divided into 4 groups:normal control group (NC group), diabetes model group (Diabetes group), ADMSCs treatment group alone (ADMSCs group) and combined treatment group of MitoQ pretreated ADMSCs (MitoQ-ADMSCs group). Diabetes models were made in mice and canines except for the NC group, the animals in the ADMSCs and MitoQ-ADMSCs groups were transplanted with 2×10⁶ cells each mice, and 1×10⁷ cells each canines once a week, for 3 consecutive weeks. Changes in clinical indicators were continuously monitored during modeling and treatment, and samples were collected 1 week after treatment and analyzed for serological levels, histological levels, oxidative stress levels and serum metabolomic changes in canines. The results showed that MitoQ treatment of ADMSCs did not affect their cell morphology, but could promote their growth, proliferation and migration ability, as well as enhance their antioxidant and anti-aging abilities. The treatment of ADMSCs and After transplantation of ADMSCs and MitoQ-ADMSCs, it was found that MitoQ significantly promoted the ability of ADMSCs to lower blood glucose, and improved pancreatic β-cell function and the body's blood glucose regulation ability. Staining of pancreatic and liver tissues revealed that ADMSCs could reduce tissue damage, promote insulin secretion, and reduce impaired hepatic glycogen synthesis and fibrosis levels, however the effects were limited, while MitoQ was able to enhance these effects more significantly. Detection of indicators related to oxidative stress in pancreatic tissues showed that MitoQ treatment of ADMSCs could effectively enhance antioxidant capacity of the body and reduce oxidative damage, thus enhancing the therapeutic effect. Metabolomic analysis of canine serum revealed that MitoQ treatment of ADMSCs significantly promoted the expression of antioxidant-related metabolites in the body. The results revealed that MitoQ could improve the antioxidant capacity of ADMSCs and accelerate the repair of tissue damage resulted from diabetes, thus beneficial for the treating of metabolism diseases in mice and canines.

Key words: mitoquionl mesylate, adipose-derived mesenchymal stem cells, canine, diabetes

中图分类号:

S856.5

谭宁, 李巴仑, 韩苗, 李琛琛, 景远翔, 寇正, 李娜, 彭莎, 赵献军, 华进联. 米托蒽醌甲磺酸盐预处理脂肪间充质干细胞对犬糖尿病的治疗效果评价[J]. 畜牧兽医学报, 2024, 55(3): 1328-1344.

TAN Ning, LI Balun, HAN Miao, LI Chenchen, JING Yuanxiang, KOU Zheng, LI Na, PENG Sha, ZHAO Xianjun, HUA Jinlian. Evaluation of Therapeutic Effect of Mitoquionl Mesylate Pretreated Adipose Derived Mesenchymal Stem Cells on Canine Diabetes Mellitus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1328-1344.

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米托蒽醌甲磺酸盐预处理脂肪间充质干细胞对犬糖尿病的治疗效果评价

Evaluation of Therapeutic Effect of Mitoquionl Mesylate Pretreated Adipose Derived Mesenchymal Stem Cells on Canine Diabetes Mellitus

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