CAJ | 학술논문

目的通过对类风湿关节炎(RA)动物模型胶原诱导性关节炎(CIA)大鼠的实验研究,观察异基因骨髓间充质干细胞(MSCs)移植对早期、晚期CIA大鼠免疫细胞和分子的免疫学作用,并探讨其在体内发挥免疫调节作用的机制.方法采用密度梯度离心结合贴壁培养法体外分离、培养大鼠MSCs,细胞表型鉴定;建立CIA大鼠模型;MSCs移植均采用尾静脉注射,移植后第42天全部处死动物取脾,通过实时定量-聚合酶链反应(RT-PCR)测定Foxp3 mRNA的表达水平,流式细胞术测定CD4+CD25+调节性T细胞的变化.采用单因素方差分析、LSD-t检验法进行统计学分析.结果早期、晚期CIA对照组CD4+CD25+调节性T细胞(1.6±0.6,1.4±0.6)和Foxp3 mRNA(0.88±0.20,0.91±0.12)的表达水平低于健康组和治疗组,差异有统计学意义(P＜0.05),早期治疗组CD4+CD25+调节性T细胞(5.0±0.4)比晚期治疗组(3.9±0.4)有所升高,差异有统计学意义(P＜0.05).结论异基因MSCs移植可通过上调CIA大鼠体内CD4+CD25+调节性T细胞水平,促进Foxp3 mRNA的表达而发挥其在体内的免疫调节作用,早期治疗组的疗效优于晚期治疗组.
목적 통과대류풍습관절염(RA)동물모형효원유도성관절염(CIA)대서적실험연구,관찰이기인골수간충질간세포(MSCs)이식대조기、만기CIA대서면역세포화분자적면역학작용,병탐토기재체내발휘면역조절작용적궤제.방법 채용밀도제도리심결합첩벽배양법체외분리、배양대서MSCs,세포표형감정;건립CIA대서모형;MSCs이식균채용미정맥주사,이식후제42천전부처사동물취비,통과실시정량-취합매련반응(RT-PCR)측정Foxp3 mRNA적표체수평,류식세포술측정CD4+CD25+조절성T세포적변화.채용단인소방차분석、LSD-t검험법진행통계학분석.결과 조기、만기CIA대조조CD4+CD25+조절성T세포(1.6±0.6,1.4±0.6)화Foxp3 mRNA(0.88±0.20,0.91±0.12)적표체수평저우건강조화치료조,차이유통계학의의(P＜0.05),조기치료조CD4+CD25+조절성T세포(5.0±0.4)비만기치료조(3.9±0.4)유소승고,차이유통계학의의(P＜0.05).결론 이기인MSCs이식가통과상조CIA대서체내CD4+CD25+조절성T세포수평,촉진Foxp3 mRNA적표체이발휘기재체내적면역조절작용,조기치료조적료효우우만기치료조.
Objective To observe the immunologic effect of transplantation of MSCs by studying the early and later period of collagen induced arthritis. Methods Rats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture plastic bottle, and the phenotypes were assessed by flow cytometry. We established collagen induced arthritis rats model. MSCs wereinjected from tail veins. We observed the expression of Foxp3 mRNA using RT-PCR, and the level of CD4+CD25+ T cell was tested by flow cytometry. One-way ANOVA and LSD-t test were used for statistical analysis.Results The percentage of CD4+ CD25+ T cells in early and later CIA groups was lower than that of normal control group and treatment groups, which showed statistically significant difference (P＜0.05). The level of CD4+ CD25+ T cell in early MSCs treatment group was higher than the later MSNs treatment group, which showed statistically significant difference (P＜0.05). Compared to the normal group and treatment groups, the expression level of Foxp3 mRNA in the early and later CIA groups was decreased markedly, while the early MSNs treatment group versus the later treatment group showed no statistically significant difference (P＞0.05).The intensity of Foxp3 mRNA in the treatment groups was similar to normal control group. Conclusion In this study, MSCs has shown significant immune-modulatory effects. It up-regulates the level of CD4+ CD25+ T cell in CIA rats, accelerate the expression of Foxp3 mRNA. The early treatment group is more effective than the late treatment group.