异种双份脐血干细胞移植对胶原诱导性关节炎小鼠IL-4及TNF-α mRNA表达的影响

Effects of double stem cells on the expression of IL-4 and TNF-α mRNA in mice with collagen-induced arthritis

目的观察异种、异基因双份人脐血间充质干细胞(MSCs)移植对Ⅱ型胶原诱导性关节炎(CIA)小鼠白细胞介素(IL)-4及肿瘤坏死因子(TNF)-α mRNA表达的影响。方法先进行脐血CD34+细胞分选,然后采用弗氏完全佐剂和Ⅱ型胶原诱导C57BL/6(H-2b)小鼠,建立CIA小鼠模型,并随机分为正常对照组、模型组、单份MSCs移植治疗组、双份MSCs移植治疗组、甲氨蝶呤阳性治疗对照组,每组10只小鼠。采用尾静脉注射,除模型组、正常对照组用生理盐水,两个MSCs移植治疗组给予的脐血所含的CD34+细胞数均为2×105。移植后第42天处死动物、取脾组织,采用反转录PCR(RT-PCR)法检测IL-4、TNF-α mRNA的表达,收集数据并做统计分析。结果双份MSCs移植治疗组IL-4、TNF-α mRNA的表达,与对照组比较,差异无统计学意义(P>0.05)。结论双份MSCs移植治疗CIA小鼠能显著抑制TNF-α mRNA的表达,上调IL-4 mRNA表达,具有免疫调节作用,可能有助于类风湿关节炎的治疗。

Objective To observe the effects of double mesenchymal stem cells（MSCs） on the expression of interleukin-4（IL-4） and tumor necrosis factor-α（TNF-α） mRNA in mice with type Ⅱ collagen-induced arthritis（CIA）.Methods CD34＋ cord blood stem cells were selected and identified.C57BL/6（H-2b） mice were induced by Freund’s complete adjuvant and type Ⅱ collagen to establish CIA mice model.All 50 mice were divided into 5 groups,including normal group,model group,mono-MSCs group,double-MSCs group and MTX group（10 mice for each）.Mice in mono-and double-MSCs groups were injected with MSCs,with the count of CD34＋ cells for 2×105,through caudal vein,and mice in normal and model group were injected with 0.9% normal saline（NS）.On the 42ed day after transplantation,mice of all groups were executed and splenic tissues were collected and detected for the expression levels of IL-4 and TNF-α mRNA by RT-PCR.All data were collected and statistically analyzed.Results There was no statistical difference of the expression levels of IL-4 and TNF-α mRNA between double-MSCs group and normal group（P〉0.05）.Conclusion Double-MSCs transplantation,which could significantly inhibit the expression of TNF-α mRNA and up-regulate the expression of IL-4 mRNA in CIA mice model,might have immune regulation effects.Double-MSCs could have the potential for the treatment of rheumatoid arthritis.