中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (6): 1058-1063.doi: 10.3969/j.issn.1673-8225.2010.06.022

• 干细胞移植 • 上一篇    下一篇

Flk-1+骨髓间充质干细胞移植上调白细胞介素6水平：是否同时加重了胶原诱导性关节炎小鼠的症状？

陈  斌，黄  姗，胡建立，孙  昭，韩  钦，宋增璇，赵春华   

  1. 北京协和医学院基础学院组织工程中心，北京市 100005
  • 出版日期:2010-02-05 发布日期:2010-02-05
  • 通讯作者: 赵春华，博士，教授，北京协和医学院基础学院组织工程中心，北京市 100005 chunhuaz@public.tpt.tj.cn
  • 作者简介:陈 斌，男，1984年生，江西省南昌市人，汉族，2009年清华大学医学部北京协和医学院毕业，博士，主要从事免疫方面的研究。cb84@163.com
  • 基金资助:

    863计划重大专项（2006AA02A109，2006AA02A115），课题名称为干细胞新药“骨髓原始间充质干细胞”Ⅱ期临床试验研究

Flk-1+ bone marrow mesenchymal stem cell transplantation upregulates interleukin-6 level: Whether it simultaneously aggravates collagen-induced arthritis in mice?

Chen Bin, Huang Shan, Hu Jian-li, Sun Zhao, Han Qin, Song Zeng-xuan, Zhao Chun-hua   

  1. Tissue Engineering Center, Institute of Basic Medical Sciences, Peking Union Medical College, Beijing   100005, China
  • Online:2010-02-05 Published:2010-02-05
  • Contact: Zhao Chun-hua, Doctor, Professor, Tissue Engineering Center, Institute of Basic Medical Sciences, Peking Union Medical College, Beijing 100005, China chunhuaz@public.tpt.tj.cn
  • About author:Chen Bin, Doctor, Tissue Engineering Center, Institute of Basic Medical Sciences, Peking Union Medical College, Beijing 100005, China cb84@163.com
  • Supported by:

    the Major 863 Project Foundation, 2006AA02A109, 2006AA02A115**

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摘要：

背景：间充质干细胞的免疫调节作用是被大家普遍认可的，在以往实验中也对Flk-1+骨髓间充质干细胞体外抑制T/B淋巴细胞增殖的能力进行了确认。

目的：验证Flk-1+骨髓间充质干细胞对胶原诱导性关节炎小鼠的治疗作用。

方法：健康10周龄雄性DBA-1(H-2Kq)小鼠18只，随机分为3组：初次免疫后细胞移植组、加强免疫后细胞移植组、模型对照组，3组小鼠均通过尾皮下注射牛Ⅱ型胶原进行初次免疫，21 d后同法进行加强免疫，建立胶原诱导性关节炎模型。密度梯度离心法和贴壁筛选法体外分离DBA-1(H-2Kq)小鼠Flk-1+骨髓间充质干细胞，初次免疫后细胞移植组小鼠在初次免疫后立即经尾静脉输注Flk-1+骨髓间充质干细胞(1~2)×106个/只，加强免疫后细胞移植组小鼠在加强免疫时同法输注等量Flk-1+骨髓间充质干细胞，模型对照组小鼠于初次免疫后0或21 d尾静脉输注等量生理盐水。观察初次免疫后和加强免疫后各组小鼠的爪垫增厚情况、临床评分，检测小鼠关节病理学变化及血清因子质量浓度的动态变化。

结果与结论：与模型对照组比较，初次免疫后细胞移植组爪垫增厚程度及平均临床疾病得分均无明显差异(P > 0.05)，均可见明显的滑膜组织损伤和炎症细胞浸润，各血清细胞因子质量浓度基本相似；而加强免疫后细胞移植组爪垫明显增厚(P < 0.01)，平均临床疾病得分高达3.35分，滑膜损伤严重，毛细血管增生，在初次免疫后28 d白细胞介素6血清浓度急剧上升(P < 0.1)，初次免疫后35 d白细胞介素6血清浓度又明显下降(P < 0.1)。提示在胶原诱导性关节炎小鼠模型中，Flk-1+骨髓间充质干细胞移植不但未取得预期的治疗效果，还在加强免疫后细胞移植组观察到显著地关节炎症状恶化现象，其可能通过上调白细胞介素6血清浓度加重类风湿关节炎小鼠的行为症状。

关键词: 免疫调节, 胶原诱导性关节炎, 白细胞介素6, Flk-1+骨髓间充质干细胞, 干细胞

Abstract:

BACKGROUND: Immunoloregulation of mesenchymal stem cells (MSCs) is commonly approved. Previous studies have confirmed the ability of Flk-1+ bone marrow MSCs (BMSCs) to inhibit T/B lymphocyte proliferation in vitro.

OBJECTIVE: To investigate the therapeutic effect of Flk-1+ BMSCs in collagen-induced arthritis mice.

METHODS: A total of 18 healthy male DBA-1(H-2Kq) mice aged 10 weeks were randomly divided into 3 groups. All the mice were injected at the base of the tail with bovine type II collagen (CII), and received a booster injection of CII on day 21 to establish the CIA mice model. DBA-1(H-2Kq)mouse Flk-1+ BMSCs were isolated in vitro by the density gradient centrifugation and adherence screening. Following initial immunity, mice in the cell transplantation group were infused with Flk-1+ BMSCs (1-2)×106 cells/mouse via the caudal vein. Mice in the cell transplantation group were injected with the same volume of Flk-1+ BMSCs during booster. Mice in the model control group were injected with an equal volume of saline 0 or 21 days following initial immunity. Following initial immunity and booster immunization, claw pad thickening and clinical score were observed, changes of joint pathology and dynamic changes in serum factor mass concentration were determined in mice. 

RESULTS AND CONCLUSION: Compared with the model control group, no significant difference in claw pad thickening and mean clinical score was detected in the cell transplantation group following initial immunity (P > 0.05), with the presence of obvious damage to synovial membrane and inflammatory cell infiltration. Mass concentration of each serum cell factor was similar. The claw pad was significantly thickened (P < 0.01), mean clinical score reached 3.35 points, with severe damage to synovial membrane, proliferation of blood capillary in the cell transplantation group following booster immunization. Interleukin-6 levels were greatly increased at day 28 following initial immunity (P < 0.1), but decreased at day 35 following initial immunity (P < 0.1). Results indicated that in the collagen-induced arthritis mouse models, Flk-1+ BMSC transplantation did not obtain prospective therapeutic efficacy, but aggravation of arthritis was observed in the cell transplantation group following booster immunization. Upregulation of interleukin-6 concentration could aggravate the behavior symptom of rheumatoid arthritis mice.

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