中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
5期
1091-1093
,共3页
张明生%穆永慧%陈清汉%朱宇
張明生%穆永慧%陳清漢%硃宇
장명생%목영혜%진청한%주우
骨髓间充质干细胞%类风湿关节炎%调节性T细胞
骨髓間充質榦細胞%類風濕關節炎%調節性T細胞
골수간충질간세포%류풍습관절염%조절성T세포
Bone marrow mesenchymal stem cells%Rheumatoid arthritis%Regulatory T cells
目的 观察间充质干细胞(MSCs)对Ⅱ型胶原诱导的关节炎(CIA)大鼠Treg细胞数量和功能的影响.方法 以Ⅱ型胶原蛋白免疫SD大鼠建立CIA动物模型;分离培养正常SD大鼠骨髓MSCs,尾静脉移注CIA大鼠;流式细胞仪检测大鼠脾脏CD4 +CD25+T细胞比例变化;实时定量聚合酶链反应(Real-time PCR)检测脾脏叉状头/翅膀状螺旋转录因子(Foxp3) mRNA水平及白细胞介素(IL)-10、转化生长因子-β1(TGF-β1)的表达;免疫磁珠法分选各组大鼠脾脏CD4+ CD25+T细胞,体外检测CD4+ CD25+T细胞对CD4+ CD25-T细胞(Teff)增殖的抑制作用.结果 MSCs移植改善了CIA大鼠的关节炎症状;与对照组比较,MSCs治疗组大鼠的脾脏内Treg细胞比例逐渐增加,到30 d时,和正常对照组比较差异无统计学意义(P＞0.05).Foxp3 mRNA水平逐渐升高,同时IL-10、TGF-β1 mRNA水平亦逐渐增加;MSCs治疗组Treg细胞对Teff细胞增殖作用与CIA组比较显著增强(P＜0.05).结论 通过移植MSCs可提高CIA大鼠体内Treg细胞的数量和功能,通过免疫调节改善了CIA大鼠关节炎症的进程.
目的 觀察間充質榦細胞(MSCs)對Ⅱ型膠原誘導的關節炎(CIA)大鼠Treg細胞數量和功能的影響.方法 以Ⅱ型膠原蛋白免疫SD大鼠建立CIA動物模型;分離培養正常SD大鼠骨髓MSCs,尾靜脈移註CIA大鼠;流式細胞儀檢測大鼠脾髒CD4 +CD25+T細胞比例變化;實時定量聚閤酶鏈反應(Real-time PCR)檢測脾髒扠狀頭/翅膀狀螺鏇轉錄因子(Foxp3) mRNA水平及白細胞介素(IL)-10、轉化生長因子-β1(TGF-β1)的錶達;免疫磁珠法分選各組大鼠脾髒CD4+ CD25+T細胞,體外檢測CD4+ CD25+T細胞對CD4+ CD25-T細胞(Teff)增殖的抑製作用.結果 MSCs移植改善瞭CIA大鼠的關節炎癥狀;與對照組比較,MSCs治療組大鼠的脾髒內Treg細胞比例逐漸增加,到30 d時,和正常對照組比較差異無統計學意義(P＞0.05).Foxp3 mRNA水平逐漸升高,同時IL-10、TGF-β1 mRNA水平亦逐漸增加;MSCs治療組Treg細胞對Teff細胞增殖作用與CIA組比較顯著增彊(P＜0.05).結論 通過移植MSCs可提高CIA大鼠體內Treg細胞的數量和功能,通過免疫調節改善瞭CIA大鼠關節炎癥的進程.
목적 관찰간충질간세포(MSCs)대Ⅱ형효원유도적관절염(CIA)대서Treg세포수량화공능적영향.방법 이Ⅱ형효원단백면역SD대서건립CIA동물모형;분리배양정상SD대서골수MSCs,미정맥이주CIA대서;류식세포의검측대서비장CD4 +CD25+T세포비례변화;실시정량취합매련반응(Real-time PCR)검측비장차상두/시방상라선전록인자(Foxp3) mRNA수평급백세포개소(IL)-10、전화생장인자-β1(TGF-β1)적표체;면역자주법분선각조대서비장CD4+ CD25+T세포,체외검측CD4+ CD25+T세포대CD4+ CD25-T세포(Teff)증식적억제작용.결과 MSCs이식개선료CIA대서적관절염증상;여대조조비교,MSCs치료조대서적비장내Treg세포비례축점증가,도30 d시,화정상대조조비교차이무통계학의의(P＞0.05).Foxp3 mRNA수평축점승고,동시IL-10、TGF-β1 mRNA수평역축점증가;MSCs치료조Treg세포대Teff세포증식작용여CIA조비교현저증강(P＜0.05).결론 통과이식MSCs가제고CIA대서체내Treg세포적수량화공능,통과면역조절개선료CIA대서관절염증적진정.
Objective To observe the immunologic effect of transplantation of mesenchymal stem cells (MSCs) on CD4+ CD25 + Treg cells frequency and function in collagen-induced arthritis (CIA) rat model.Methods Rat CIA model was established by immunizing SD rats with type Ⅱ collagen.MSCs were purified and cultured from the bone marrow of SD rats,then transplanted to CIA model.The frequency of CD4 + CD25 + Treg cells in the spleen was measured by flow cytometry (FCM) respectively after the MSCs transplantation,and the expression of forkhead box P3 (Foxp3),transforming growth factor-β1 (TGF-β1)and interleukin (IL)-10 mRNA was evaluated by real-time quantitative polymerase chain reaction (Real-time PCR).CD4 + CD25 + Treg cells were isolated from the spleen of rats by immunomagnetic beads and the inhibitory effect of CD4 + CD25 + Treg cells on proliferation of CD4 + CD25-Teff cells was estimated by methyl thiazol tetrazolium (MTT) colorimetry.Results The MSCs transplantation relieved the arthritis symptom of the CIA rats.The frequency of CD4 + CD25 + Treg cells in the spleen of MSCs-treated CIA rats was increased significantly (P ＜ 0.05) as compared with the CIA group,and the levels of Foxp3,IL-10 and TGF-β1 mRNA were also significantly increased (P ＜ 0.05).CD4 + CD25 + Treg cells isolated from treatment group showed higher inhibition on the proliferation of CD4 + CD25-Teff cells versus CD4 + CD25 +Treg cells isolated from CIA rats.Conclusion Transplantation of MSCs is effective on preventing the de-velopment of CIA.The increased frequency and reinforced function of CD4+ CD25 + Treg cells may be one of the mechanisms for MCSs transplantation treating CIA in rats.
