人脐带间充质干细胞移植治疗系统性红斑狼疮

文章亮点： 1 传统的治疗方案，如糖皮质激素联合免疫抑制剂治疗，虽能有效提高系统性红斑狼疮患者的长期存活率， 但部分患者仍然复发，部分患者治疗无效，仍然有潜在的致死率。理论上间充质干细胞可用于治疗系统性红 斑狼疮。 2 C57BL/lpr 实验小鼠移植人脐带间充质干细胞治疗 3 次后，外周血抗核抗体、抗组蛋白抗体、抗双链 DNA 抗体均明显下降，调节性 T 细胞阳性率、Foxp3 细胞阳性率均升高，OPG 和 Foxp3 基因表达也明显升高， 接近正常对照组，表明 C57BL/lpr 小鼠免疫细胞变化趋向正常。 关键词： 干细胞；移植；系统性红斑狼疮；人脐带间充质干细胞；C57BL/lpr 小鼠 主题词： 脐带；间质干细胞移植；红斑狼疮, 系统性 基金资助： 国家支撑计划(2014BAI01B01)；云南省科技计划项目(2013CA005)
阮光萍，姚翔，刘菊芬，王金祥，胡媛媛，李自安，杨建勇，庞荣清，潘兴华. 人脐带间充质干细胞移植治疗 系统性红斑狼疮[J].中国组织工程研究，2015，19(14):2172-2178.
doi:10.3969/j.issn.2095-4344.2015. 14.008
Transplantation of human umbilical cord mesenchymal stem cells in the treatment of systemic lupus erythematosus
Ruan Guang-ping, Yao Xiang, Liu Ju-fen, Wang Jin-xiang, Hu Yuan-yuan, Li Zi-an, Yang Jian-yong, Pang Rong-qing, Pan Xing-hua (The Cell Biological Therapy Center, Kunming General Hospital of Chengdu Military Command, Kunming 650032, Yunnan Province, China)
Accepted: 2015-03-06
2172
P.O. Box 10002, Shenyang 110180 www.CRTER.org
阮光萍，等. 人脐带间充质干细胞移植治疗系统性红斑狼疮
the end of treatment, blood samples were collected to measure antinuclear antibody, anti-histone antibody, anti-double stranded DNA antibody changes; OPG and Foxp3 gene expression changes were detected by quantitative PCR method. RESULTS AND CONCLUSION: After treatment, the levels of anti-nuclear antibodies, anti-histone antibodies and anti-double stranded DNA antibodies in the peripheral blood of mice were all declined in the low-, medium- and high-dose groups, while the number of peripheral blood CD4+CD25+T cells was significantly elevated. OPG and Foxp3 gene expression was also increased dramatically in the low-, medium- and high-dose groups, which was similar to that in the normal control group and significantly different from that in the model control group (P < 0.01). Experimental findings demonstrate that after transplantation of human umbilical cord mesenchymal stem cells, all relevant indicators in C57BL/lpr mice recovered to the normal levels, and the high-dose treatment group had the most obvious effect.
摘要 背景：系统性红斑狼疮是一种以多器官或多系统病变和血清中出现多种自身抗体为特征的自身免疫性疾病， 目前缺乏有效的治疗方案，而理论上间充质干细胞可用于治疗系统性红斑狼疮。 目的：观察人脐带间充质干细胞移植治疗系统性红斑狼疮小鼠的疗效。 方法：分离培养人脐带间充质干细胞，并用深红色荧光 DiR 标记细胞。实验小鼠分 5 组：正常对照组(C57BL 小鼠)，模型对照组(C57BL/lpr 小鼠)，低、中、高剂量脐带间充质干细胞治疗组(C57BL/lpr 小鼠)，每组 10 只。 各治疗组通过尾静脉注射低、中、高剂量(2×106，1×106，0.5×106 个)脐带间充质干细胞，每周 1 次，连续 3 周，治疗结束采血测抗核抗体、抗组蛋白抗体、抗双链 DNA 抗体变化，定量 PCR 检测 OPG 和 Foxp3 基因 表达的变化。 结果与结论：细胞移植 3 次后，外周血抗核抗体、抗组蛋白抗体、抗双链 DNA 抗体均明显下降，CD4+CD25+T 细胞明显升高，OPG 和 Foxp3 基因表达也明显升高，接近正常对照组，与模型对照组相比差异均有显著性意 义(P < 0.01)。结果表明人脐带间充质干细胞能使 C57BL/lpr 小鼠的各项相关指标恢复到 C57BL 正常鼠水平， 以高剂量治疗组效果最明显。
Biblioteka Baidu
Ruan Guang-ping, M.D., Associate chief physician, the Cell Biological Therapy Center, Kunming General Hospital of Chengdu Military Command, Kunming 650032, Yunnan Province, China
阮光萍，女，1974 年生， 云南省昆明市人，汉族， 2007 年 南方医 科大学毕 业，博士，副主任医师， 主要从事干细胞基础与临 床研究。
通讯作者：潘兴华，博士， 主任医师，解放军成都军 区昆明总医院细胞生物治 疗中心，云南省昆明市 650032
中图分类号:R394.2 文献标识码:B 文章编号:2095-4344 (2015)14-02172-07 稿件接受：2015-03-06 http://WWW.crter.org
Abstract BACKGROUND: Systemic lupus erythematosus is an autoimmune disease characterized as an emergence of a variety of autoantibodies in serum and multi-system and multi-organ lesions. Currently, there is a lack of effective treatment options, and umbilical cord mesenchymal stem cells are a promising therapy for systemic lupus erythematosus based on cell biological roles. OBJECTIVE: To observe the therapeutic efficacy of human umbilical cord mesenchymal stem cell transplantation in the treatment of systemic lupus erythematosus in mice. METHODS: Human umbilical cord mesenchymal stem cells were isolated and cultured followed by labeling with DiR fluorescence. Experimental mice were divided into normal control group (C57BL mice), model control group (C57BL/lpr mice), low-, medium- and high-dose umbilical cord mesenchymal stem cell therapy groups (C57BL/lpr mice), with 10 mice in each group. Mice in the low-, medium- and high-dose groups were respectively injected 0.5×106, 1×106, 2×106 human umbilical cord mesenchymal stem cells, once a week, for 3 consecutive weeks. At
Corresponding author: Pan Xing-hua, M.D., Chief physician, the Cell Biological Therapy Center, Kunming General Hospital of Chengdu Military Command, Kunming 650032, Yunnan Province, China
Ruan GP, Yao X, Liu JF, Wang JX, Hu YY, Li ZA, Yang JY, Pang RQ, Pan XH. Transplantation of human umbilical cord mesenchymal stem cells in the treatment of systemic lupus erythematosus. Zhongguo Zuzhi Gongcheng Yanjiu. 2015;19(14):2172-2178.
www.CRTER.org
0 引言 Introduction
系统性红斑狼疮是一个累及身体多系统多器官的自身 免疫性疾病，包括肾、心血管、神经、骨骼肌和皮肤系统[1]。 尽管免疫抑制药物的使用能提高其长期存活率，但仍然有 潜在的致死率，特别是对于抵抗治疗的患者。系统性红斑 狼疮的病理过程包括破坏目标器官和组织，累积自身反应 性淋巴细胞和免疫复合物。尽管系统性红斑狼疮患者病情 的严重程度和受累的器官有明显不同，但广泛存在T、B淋 巴细胞的异常[1-3]。而且，系统性红斑狼疮显示出多面性的 免疫调节效应，包括免疫缺陷和免疫超反应。深入理解系 统性红斑狼疮的基本病理机制是治疗的关键，可恢复免疫 稳态而不影响对病原的保护性免疫反应[4]。
中国组织工程研究 第 19 卷 第 14 期 2015–04–02 出版 Chinese Journal of Tissue Engineering Research April 2, 2015 Vol.19, No.14
人脐带间充质干细胞移植治疗系统性红斑狼疮
www.CRTER.org
阮光萍，姚 翔，刘菊芬，王金祥，胡媛媛，李自安，杨建勇，庞荣清，潘兴华(解放军成都军区昆明总医院细胞生物治疗中心，云南省 昆明市 650032)
Subject headings: Umbilical Cord; Mesenchymal Stem Cell Transplantation; Lupus Erythematosus, Systemic Funding: the National Science and Technology Support Plan of China, No. 2014BAI01B01; the Science and Technology Plan Project of Yunnan Province, No. 2013CA005