CAJ | 학술논문

目的：评价人源脂肪干细胞对辐射诱导的肠血管损伤的修复作用。方法选用成年雄性SD大鼠，共34只，给予全腹部15 Gy X射线照射。造模后，取其中17只给予P6代人源脂肪干细胞腹腔注射治疗（ Ad-MSC治疗组）；另17只大鼠给予双磷酸盐缓冲液（ PBS）腹腔注射治疗（ PBS溶剂对照组）。照射后第10天，流式细胞分析绒毛内CD31阳性内皮细胞的数量，免疫荧光染色分析新生的血管内皮细胞，免疫组织化学染色分析血管结构的连续性；并提取受照小肠组织总RNA，实时荧光定量PCR检测受照组织内基质细胞起源因子-1（SDF-1）、血管内皮细胞生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）以及血管内皮细胞生长因子受体（Flk-1）的表达量。检测Ad-MSC治疗组的受损小肠内新生的内皮细胞来源。结果与PBS溶剂对照组相比，Ad-MSC能够显著增加受损组织内CD31阳性内皮细胞数量（t＝12?15，P＜0?05），提高受损组织内的血管密度（20 d：t＝10?33，P＜0．05；30 d：t＝32?85，P＜0?05），上调血管生成基因VEGF、bFGF、Flk-1以及SDF-1的表达量（t＝10?34、11?25、6?73、6?73，P＜0?05）。维持受损部位小肠绒毛内的血管完整，并促进CD31阳性造血干/祖细胞向血管内皮细胞的分化，加速受损部位血管的新生。结论人源脂肪干细胞通过发挥促血管新生的作用来修复辐射诱导的肠血管损伤。
목적：평개인원지방간세포대복사유도적장혈관손상적수복작용。방법선용성년웅성SD대서，공34지，급여전복부15 Gy X사선조사。조모후，취기중17지급여P6대인원지방간세포복강주사치료（ Ad-MSC치료조）；령17지대서급여쌍린산염완충액（ PBS）복강주사치료（ PBS용제대조조）。조사후제10천，류식세포분석융모내CD31양성내피세포적수량，면역형광염색분석신생적혈관내피세포，면역조직화학염색분석혈관결구적련속성；병제취수조소장조직총RNA，실시형광정량PCR검측수조조직내기질세포기원인자-1（SDF-1）、혈관내피세포생장인자（VEGF）、감성성섬유세포생장인자（bFGF）이급혈관내피세포생장인자수체（Flk-1）적표체량。검측Ad-MSC치료조적수손소장내신생적내피세포래원。결과여PBS용제대조조상비，Ad-MSC능구현저증가수손조직내CD31양성내피세포수량（t＝12?15，P＜0?05），제고수손조직내적혈관밀도（20 d：t＝10?33，P＜0．05；30 d：t＝32?85，P＜0?05），상조혈관생성기인VEGF、bFGF、Flk-1이급SDF-1적표체량（t＝10?34、11?25、6?73、6?73，P＜0?05）。유지수손부위소장융모내적혈관완정，병촉진CD31양성조혈간/조세포향혈관내피세포적분화，가속수손부위혈관적신생。결론인원지방간세포통과발휘촉혈관신생적작용래수복복사유도적장혈관손상。
Objective To assess the therapeutic effect of human adipose-derived mesenchymal stem cells on radiation-induced vascular injury in the small intestine of rat. Methods A total of 34 male Sprague-Dawley rats were enrolled in this study. To establish a model of radiation-induced intestinal injury, each rat was irradiated with 15 Gy in whole abdomen. 17 rats were randomly selected and infused intraperitoneally with passage 6 ( P6 ) Ad-MSCs, and the other 17 rats that received PBS were set as control. 10 days post-irradiation, the number of CD31+ endothelial cells in the small intestine villus was measured by flow-cytometry, the expressions of CD31, CD105 and isolectin-B4 in the na?ve endothelial cells with detected by IHC-staining, and the vascular integrity was evaluated by measuring VE-Cadherin. The origination of na?ve endothelial cells within injured intestine was also analyzed. In addition, total mRNA were extracted from irradiated small intestine to assay the expressions of VEGF, bFGF, Flk-1 and SDF-1 using quantitative Real-time PCR. Results Compared to the control, the amount of CD31-postive endothelial cells within irradiated intestine was significantly increased after Ad-MSCs infusion ( t=12?15, P<0?05). The microvascular density in the injured sites was also significantly increased by the infusion of Ad-MSCs (20 d:t=10?33, P<0. 05;30 d:t=32?85, P<0?05). Moreover, the expressions of VEGF, bFGF, Flk-1 and SDF-1 were significantly up-regulated after delivery of Ad-MSCs ( VEGF:t =10?34, bFGF:t=11?25,Flk-1:t=6?73, SDF-1:t=6?73, all P<0?05), which was beneficial in maintaining the integrity of intra-villus blood-vessels as well as promoting neovascularization in the injured sites. Conclusion Ad-MSCs had potentials in healing radiation-induced vascular injury in rat small intestine.