CAJ | 학술논문

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-2K~q)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-2K~q)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)×10~6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.
배경:간충질간세포적면역조절작용시피대가보편인가적,재이왕실험중야대Flk-1~+골수간충질간세포체외억제T/B림파세포증식적능력진행료학인.목적:험증Flk-1~+골수간충질간세포대효원유도성관절염소서적치료작용.방법:건강10주령웅성DBA-1(H-2K~q)소서18지,수궤분위3조:초차면역후세포이식조、가강면역후세포이식조、모형대조조,3조소서균통과미피하주사우Ⅱ형효원진행초차면역,21 d후동법진행가강면역,건립효원유도성관절염모형.밀도제도리심법화첩벽사선법체외분리DBA-1(H-2K~q)소서Flk-1~+골수간충질간세포,초차면역후세포이식조소서재초차면역후립즉경미정맥수주Flk-1~+골수간충질간세포(1～2)×10~6개/지,가강면역후세포이식조소서재가강면역시동법수주등량Flk-1~+골수간충질간세포,모형대조조소서우초차면역후0혹21 d미정맥수주등량생리염수.관찰초차면역후화가강면역후각조소서적조점증후정황、림상평분,검측소서관절병이학변화급혈청인자질량농도적동태변화.결과여결론:여모형대조조비교,초차면역후세포이식조조점증후정도급평균림상질병득분균무명현차이(P > 0.05),균가견명현적활막조직손상화염증세포침윤,각혈청세포인자질량농도기본상사;이가강면역후세포이식조조점명현증후(P < 0.01),평균림상질병득분고체3.35분,활막손상엄중,모세혈관증생,재초차면역후28 d백세포개소6혈청농도급극상승(P < 0.1),초차면역후35 d백세포개소6혈청농도우명현하강(P < 0.1).제시재효원유도성관절염소서모형중,Flk-1~+골수간충질간세포이식불단미취득예기적치료효과,환재가강면역후세포이식조관찰도현저지관절염증상악화현상,기가능통과상조백세포개소6혈청농도가중류풍습관절염소서적행위증상.
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-2K~q) 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-2K~q)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.