CAJ | 학술논문

目的评价脂肪干细胞对放射性肠炎的治疗作用.方法选用成年雄性SD大鼠,共52只.随机数字表法选取46只给予全腹15 Gy X射线照射制作放射性肠炎动物模型.造模结束后2h,随机数字表法从受照大鼠中选取23只受照大鼠给予P6代青年女性脂肪干细胞腹腔注射治疗(Ad-MSC治疗组);取另23只受照大鼠给予双磷酸盐缓冲液(PBS)腹腔注射治疗(PBS溶剂对照组);剩余6只未照射大鼠作为健康对照组.造模后首个10d内,分次抽取受照大鼠外周血,ELISA法检测血清中白介素-10(IL-10)的含量;分离受照大鼠胸腺细胞及外周血单个核细胞(PBMC),流式细胞术检测CD4/CD25/Foxp(3)阳性调节性T细胞的百分含量;获取受照小肠组织,HE染色分析炎症细胞浸润程度,Masson三色染色分析胶原成分的沉积程度,髓系过氧化物酶(MPO)免疫组织化学染色分析浸润的炎症细胞类型.Kaplan-Meier方法对受照大鼠进行生存分析.结果在照射后30 d内,与PBS腹腔注射治疗的大鼠相比,接受脂肪干细胞治疗的大鼠的生存时间显著延长(t=4.53,P ＜0.05).另外,脂肪干细胞能够上调受照大鼠外周血中IL-10的含量(7 d:t=13.93,P＜0.05);上调外周血(3.5 d:t =7.72,7 d:t =11.11,10 d:t=6.99,P＜0.05)和胸腺(7 d:t =16.17,10 d:t=12.12,P＜0.05) CD4/CD25/Foxp(3)阳性调节性T细胞的百分比;进而减轻炎症细胞在受照小肠中的浸润及胶原的沉积.结论脂肪干细胞对放射性肠炎具有治疗作用.
목적 평개지방간세포대방사성장염적치료작용.방법 선용성년웅성SD대서,공52지.수궤수자표법선취46지급여전복15 Gy X사선조사제작방사성장염동물모형.조모결속후2h,수궤수자표법종수조대서중선취23지수조대서급여P6대청년녀성지방간세포복강주사치료(Ad-MSC치료조);취령23지수조대서급여쌍린산염완충액(PBS)복강주사치료(PBS용제대조조);잉여6지미조사대서작위건강대조조.조모후수개10d내,분차추취수조대서외주혈,ELISA법검측혈청중백개소-10(IL-10)적함량;분리수조대서흉선세포급외주혈단개핵세포(PBMC),류식세포술검측CD4/CD25/Foxp(3)양성조절성T세포적백분함량;획취수조소장조직,HE염색분석염증세포침윤정도,Masson삼색염색분석효원성분적침적정도,수계과양화물매(MPO)면역조직화학염색분석침윤적염증세포류형.Kaplan-Meier방법대수조대서진행생존분석.결과 재조사후30 d내,여PBS복강주사치료적대서상비,접수지방간세포치료적대서적생존시간현저연장(t=4.53,P ＜0.05).령외,지방간세포능구상조수조대서외주혈중IL-10적함량(7 d:t=13.93,P＜0.05);상조외주혈(3.5 d:t =7.72,7 d:t =11.11,10 d:t=6.99,P＜0.05)화흉선(7 d:t =16.17,10 d:t=12.12,P＜0.05) CD4/CD25/Foxp(3)양성조절성T세포적백분비;진이감경염증세포재수조소장중적침윤급효원적침적.결론 지방간세포대방사성장염구유치료작용.
Objective To evaluate the therapeutic effect of adipose-derived mesenchymal stem cells on radiation enteritis.Methods A total of 52 male Sprague-Dawley rats were used in the present study.Herein,46 rats were randomly selected and irradiated with a dose of 15 Gy at their abdomens.Two hours post-irradiation,23 rats were randomly selected and infused intraperitoneally with adipose-derived mesenchymal stem cells in passage 6 from young-female donor.The other 23 rats were intraperitoneally infused with PBS.The rest 6 rats were set as normal control.During the first 10 days post-irradiation,peripheral blood-samples from irradiated rats were harvested for testing the levels of IL-10 in serum using ELISA assay.Additionally,after isolating the thymic cells and peripheral blood mononuclear cells,the percentages of CD4/CD25/Foxp(3)-positive regulatory T cells in thymus and peripheral blood were tested by flow-cytometry.Finally,infiltration of inflammatory cells and deposition of collagens within irradiated small intestine were analyzed by H&E staining and Masson Trichrome staining,respectively.Based on the MPO-immunohistochemistry staining,the type of infiltrated cells was identified.The Kaplan-Meier method was used for analyzing the survival rate of irradiated rats.Results During a period of 30 days post-irradiation,the irradiated rats receiving adipose-derived mesenchymal stem cells survived longer than those receiving PBS (t =4.53,P ＜ 0.05).Compared to the irradiated rats with PBS-treatment,adipose-derived mesenchymal stem cells could elevate the level of IL-10 in serum (7 d:t =13.93,P ＜ 0.05) and increase the percentages of CD4/CD25/Foxp(3)-positive regulatory T cells in both peripheral blood (3.5 d:t =7.72,7 d:t=11.11,10 d:t =6.99,P ＜0.05) and thymus (7 d:t =16.17,10 d:t =12.12,P＜ 0.05).Moreover,infiltration of inflammatory cells and deposition of collagens within irradiated small intestine were mitigated by adipose-derived mesenchymal stem cells.Conclusions Adipose-derived mesenchymal stem cells were capable of curing radiation enteritis.