BMSCs移植通过T细胞免疫抑制缓解1型糖尿病小鼠发病实验研究

doi:10.13418/j.issn.1001-165x.2020.06.009

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (6): 668-673.doi: 10.13418/j.issn.1001-165x.2020.06.009

BMSCs移植通过T细胞免疫抑制缓解1型糖尿病小鼠发病实验研究

余佳珊，　杨文江，　彭丽娟，　高杰，　李红，　苏敏，　胡蓉

贵州医科大学基础医学院组织学与胚胎学教研室，贵阳 550025

收稿日期:2020-03-23 出版日期:2020-11-25 发布日期:2020-12-08
通讯作者: 胡蓉，教授，硕士生导师，E-mail: hurong@gmc.edu.cn；苏敏，教授，博士生导师，E-mail: sumin@gmc.edu.cn
作者简介:余佳珊（1995-），女，四川自贡人，在读硕士，研究方向：干细胞与自身免疫病，Tel：13696026121，E-mail: 939360286@qq.com
基金资助:
贵州省科学技术基金项目[黔科合基础（2016）1125号] ；贵州省科技支撑计划（黔科合支撑 [2017]2970号）；贵州省普通高校科技拔尖人才支持计划（黔教合KY字[2017]071）；贵州省科技合作计划项目（黔科合LH字[2016]7363）；国家自然科学基金（NSFC 81660033）

Bone marrow mesenchymal stem cells transplantation attenuates type 1 diabetes mellitus by immune suppressing T cells

YU Jia-shan, YANG Wen-jiang, PENG Li-juan, GAO Jie, LI Hong, SU Min, HU Rong

Department of Human Histology and Embryology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, Guizhou Province, China

Received:2020-03-23 Online:2020-11-25 Published:2020-12-08

摘要/Abstract

摘要： 目的　探讨小鼠骨髓间充质干细胞（Bone marrow mesenchymal stem cells, BMSCs）移植缓解1型糖尿病小鼠（T1DM）发病及机制。方法　BMSCs分离培养及鉴定；第3代培养的上清作为条件培养基，与2周龄NOD小鼠脾细胞共培养，FACS检测CD4+和CD8+T细胞增殖及活化；2周龄雌性NOD小鼠随机分为BMSCs组、PBS组、对照组，每组6只，对照组2周处死；BMSCs组和PBS组12周分别腹腔注射100 μl（2×107细胞）BMSCs或等体积PBS，监测小鼠血糖和体重至26周；HE、免疫组化、免疫荧光染色观察胰腺炎性细胞浸润；FACS检测CD4+和CD8+T细胞增殖及活化。结果　成功分离培养BMSCs；BMSCs条件培养基抑制CD4+及CD8+T细胞的增殖及活化（P<0.05）；BMSCs组小鼠体重高于PBS组（P<0.05），糖尿病发病率降低，炎性浸润减少（P<0.05），脾脏CD4+和CD8+T细胞增殖及活化降低（P<0.05）。结论　BMSCs移植可能通过T细胞免疫抑制缓解T1DM小鼠发病。

关键词: 骨髓间充质干细胞, 　NOD/ShiLtJ小鼠, 　1型糖尿病, 　免疫抑制, 　T细胞

Abstract: Objective　To investigate the effects and mechanism of bone marrow mesenchymal stem cells (BMSCs) transplantation on Type 1 diabetes mellitus (T1DM) mice.　Methods　Isolation, culture and identification of BMSCs were performed. BMSCs supernatant was used to be the conditioned medium after third passage (P3), co-cultured with 2 weeks NOD splenocytes, and the proliferation and activation of CD4+ and CD8+T cells were detected by FACS. Two weeks female NOD mice were randomly divided into three groups: a BMSCs group, a PBS group and a control group (6 mice in each group). Mice in the control group were killed at 2 weeks. 100 μl (2×107 cells) BMSCs or equal volume PBS was injected intraperitoneally at 12 weeks in the BMSCs group and the PBS group respectively, then the mice were monitored for blood glucose and body weight till 26 weeks. Inflammatory cells infiltration of pancreases was detected by H&E, immunohistochemical and immunofluorescent staining. The splenocytes were extracted to observe the proliferation and activation of CD4+ and CD8+T cells by FACS.　Results　BMSCs were successfully isolated and cultured. BMSCs conditioned medium inhibited the proliferation and activation of CD4+ and CD8+T cells (P<0.05). Mice body weight of BMSCs group was higher than that of PBS group (P<0.05), the incidence rate of diabetes and inflammatory infiltration decreased (P<0.05) as well as the proliferation and activation of CD4+ and CD8+ T cells in the spleen (P<0.05).　Conclusions　BMSCs transplantation may attenuate T1DM by immune suppressing T cells.

Key words: BMSCs, 　NOD/ShiLtJ mice, 　T1DM Immunosuppression, 　T cells

中图分类号:

R329

余佳珊, 杨文江, 彭丽娟, 高杰, 李红, 苏敏, 胡蓉. BMSCs移植通过T细胞免疫抑制缓解1型糖尿病小鼠发病实验研究[J]. 中国临床解剖学杂志, 2020, 38(6): 668-673.

YU Jia-shan, YANG Wen-jiang, PENG Li-juan, GAO Jie, LI Hong, SU Min, HU Rong. Bone marrow mesenchymal stem cells transplantation attenuates type 1 diabetes mellitus by immune suppressing T cells[J]. Chinese Journal of Clinical Anatomy, 2020, 38(6): 668-673.

参考文献 15

[1]	Medina A, Parween S, Ullsten S, et al. Early deficits in insulin secretion, beta cell mass and islet blood perfusion precede onset of autoimmune type 1 diabetes in BioBreeding rats[J]. Diabetologia, 2018, 61(4):896-905.
[2]	Sims EK, Dimeglio LA. Cause or effect? A review of clinical data demonstrating beta cell dysfunction prior to the clinical onset of type 1 diabetes[J]. Molecular Metabolism, 2019, 27(suppl): S129-S138.
[3]	Xv J, Ming Q, Wang X, et al. Mesenchymal stem cells moderate immune response of type 1 diabetes[J]. Cell Tissue Res, 2017, 368(2): 239-248.
[4]	Gülden E, Palm N, Herold KC. MAIT Cells: A Link between Gut Integrity and Type 1 Diabetes[J]. Cell Metabolism, 2017, 26(6): 813-815.
[5]	Boháčová P, Holáň V. Mesenchymal stem cells and type 1 diabetes treatment[J]. Vnitrni Lekarstvi, 2018, 64(7-8): 725-728.
[6]	Liang C, Jiang E, Yao J, et al. Interferon-γ mediates the immunosuppression of bone marrow mesenchymal stem cells on T-lymphocytes in vitro[J]. Hematology, 2018, 23(1): 44-49.
[7]	Andrew SN, Massimo M, Jeffrey RA, et al. Oral therapy with colonization factor antigen I prevents development of type 1 diabetes in Non-obese Diabetic mice[J]. Scientific Reports, 2020, 10(1): 1-15.
[8]	Tian X, Lin Y, Cui C, et al. BTNL2-Ig Protein Attenuates Type 1 Diabetes in Non-Obese Diabetic (NOD) Mice[J]. Advanced healthcare materials, 2019, 8(9): 1-7.
[9]	Jamal MA, Bahare N, Shima R, et al. Immunomodulatory and protective effects of adipose tissue-derived mesenchymal stem cells in an allograft islet composite transplantation for experimental autoimmune type 1 diabetes[J]. Immunol Lett, 2017, 188(17): 21-31.
[10]	廖文萍, 胡蓉, 黄悦, 等. 骨髓间充质干细胞异体移植对小鼠实验性自身免疫性脑脊髓炎的治疗作用[J]. 解剖学报, 2017, 48(3): 266-272.
[11]	Zhang W, Zhou L, Dang J, et al. Human gingiva-derived mesenchymal stem cells ameliorate streptozoticin-induced T1DM in mice via suppression of T effector cells and up-regulating treg subsets[J]. Sci Rep, 2017, 7(1): 1-12.
[12]	Relation T, Yi T, Guess AJ, et al. Intratumoral delivery of interferonc-secreting mesenchymal stromal cells repolarizes Tumor-associated macrophages and suppresses neuroblastoma proliferation in vivo[J]. Stem Cells, 2018, 36(6): 915-924.
[13]	Lee HJ, Kim SN, Jeon MS, et al. ICOSL expression in human bone marrow-derived mesenchymal stem cells promotes induction of regulatory T cells[J]. Sci Rep, 2017, 7(7): 1-15.
[14]	Wu X, Wang Y, Xu J, et al. MM-BMSCs induce naïve CD4+ T lymphocytes dysfunction through fibroblast activation protein α[J]. Oncotarget, 2017, 8(32): 52614-52628.
[15]	Reddy S, Zeng N, Al-Diery H, et al. Analysis of peri-islet CD45-positive leucocytic infiltrates in long-standing type 1 diabetic patients: additional data regarding cause of death[J]. Diabetologia, 2015, 58(8): 1959-1959.

BMSCs移植通过T细胞免疫抑制缓解1型糖尿病小鼠发病实验研究

Bone marrow mesenchymal stem cells transplantation attenuates type 1 diabetes mellitus by immune suppressing T cells

可视化

摘要/Abstract

引用本文

使用本文

参考文献 15

相关文章 13

Metrics

本文评价

推荐阅读 0

[1]	赵仁礼, 赖国华, 吴家昌, 吴铭杰, 庄伟达, 欧阳钧, 桑宏勋. 差异超速离心法对不同种属间充质干细胞外泌体分离的可靠性研究[J]. 中国临床解剖学杂志, 2022, 40(3): 292-296.
[2]	范光碧,先德海,邓莉,汤华军,张雁儒,杨朝鲜. 电针刺激联合BMSCs移植对脑出血大鼠出血灶周边区水通道蛋白4表达的影响[J]. 中国临床解剖学杂志, 2016, 34(6): 655-660.
[3]	杨青,秦书俭,包翠芬,单伟. 两种示踪方式的骨髓间充质干细胞在促进肝缺血再灌注损伤修复中的对比研究[J]. 中国临床解剖学杂志, 2016, 34(3): 312-317.
[4]	陈杨葭,张艳金,刘佳,孙天胜,华文熙,钟剑峰,冯杰扬,叶超群. 骨髓间充质干细胞对髋部骨折诱导的老年大鼠急性肺损伤的保护机制[J]. 中国临床解剖学杂志, 2015, 33(6): 672-676.
[5]	桑遥彩,包翠芬,武蕾,张苗苗,王艳,秦书俭. 辛伐他汀对BMSCs成骨分化过程中TGF-β1/ smad信号通路的影响[J]. 中国临床解剖学杂志, 2015, 33(4): 434-438.
[6]	李庆庆,王大平,熊建义,黄江鸿,付志杰. Runx2重组慢病毒感染骨髓间充质干细胞并促进其成骨分化的研究[J]. 中国临床解剖学杂志, 2015, 33(3): 311-315.
[7]	张伟才, 黄继锋, 严琼娇, 李世普. PNGF导管复合骨髓间充质干细胞修复大鼠12 mm坐骨神经缺损[J]. 中国临床解剖学杂志, 2014, 32(3): 325-329.
[8]	邹学军, 汪金生, 左昕, 米海鹏. BMP-2通过诱导RBP4表达调控小鼠骨髓间充质细胞的成骨分化[J]. 中国临床解剖学杂志, 2014, 32(2): 184-188.
[9]	李季, 秦书俭, 包翠芬, 桑瑶采, 屈惠莹. ZHX3基因转染BMSCs及对体外成骨能力的实验研究[J]. 中国临床解剖学杂志, 2014, 32(2): 174-178.
[10]	王海萍, 张雷, 赵静, 吴志刚, 吕洋, 王浩宇, 邵素??. 大鼠骨髓间充质干细胞移植治疗心肌梗死的实验研究[J]. 中国临床解剖学杂志, 2012, 30(2): 209-213.
[11]	孙丽, 王雪儿, 张敏, 陈世萱, 孔亚男, 崔梦卿, 张璐, 张琳. Activin B联合BMSCs不同移植方式对大鼠皮肤创伤愈合的治疗[J]. 中国临床解剖学杂志, 2012, 30(2): 197-202.
[12]	邱家琦, 张鸣生, 李新平, 白文芳, 吴博, 白利明, 徐武华. 低频电磁场对BDNF基因修饰的BMSCs增殖影响的实验研究[J]. 中国临床解剖学杂志, 2012, 30(1): 74-78.
[13]	张　凯, 王大平, 朱伟民, 刘建全. 骨髓间充质干细胞与纳米羟基磷灰石支架材料的体外相容性研究[J]. 中国临床解剖学杂志, 2011, 29(2): 213-216.