肝孤核受体激动剂对间充质干细胞的抗缺氧复氧损伤作用及机制  

Liver X receptor agonist prevents adipose-derived mesenchymal stem cells from hypoxia reoxygenation injury

作　　者：李春红[1] 李霜[1] 王亚斌[1] 范伟伟[1] 梁栋[1] 李聪叶[1] 曹丰[1] 

机构地区：[1]第四军医大学西京医院心内科,西安710032

出　　处：《中华老年心脑血管病杂志》2012年第1期72-75,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的探讨肝孤核受体(LXR)激动剂对脂肪间充质干细胞(AD-MSCs)缺氧损伤的保护作用及可能机制。方法酶消化法分离稳定表达萤火虫荧光素酶(Fluc)报告基因的小鼠AD-MSCs,流式细胞术检测CD90、CD44、CD34、CD45细胞表面标记物。3代AD-MSCs分为7组:对照组;缺氧6 h/复氧2 h组(缺氧复氧组);缺氧/复氧+DMSO组(DMSO组);缺氧复氧+不同浓度LXR激动剂T0901317干预组(1μmol/L组、5μmol/L组、10μmol/L组、15μmol/L组)。运用Fluc报告基因生物发光成像技术对AD-MSCs细胞增殖进行定量评价,免疫印迹法检测细胞NF-κB表达水平,ELISA法检测AD-MSCs的白细胞介素6(IL-6)、TNF-α分泌水平。结果流式细胞术结果显示.AD-MSCs呈CD44(+)、CD90(+)、CD34(-)、CD45(-)。光学成像结果显示,AD-MSCs细胞数与Fluc平均生物发光信号强度呈正相关(r^2=0.97)。与对照组比较,缺氧复氧组AD-MSCs分泌TNF-α、IL-6、NF-κB明显升高;与缺氧复氧组比较,10μmol/L组AD-MSCs分泌TNF-α、IL-6、NF-κB明显减少(P<0.05,P<0.01)。结论 LXR激动剂可以促进缺氧复氧损伤后AD-MSCs的存活,并能通过NF-κB信号途径抑制炎性因子IL-6、TNF-α的释放,可为干细胞移植治疗心肌缺血再灌注损伤提供新的细胞保护方法。Objective To investigate the effects and possible mechanisms of liver X receptor（LXR） agonist on adipose-derived mesenchymal stem cells （AD-MSCs） after hypoxia reoxygenation （H/R） injury. Methods AD-MSCs stably expressing firefly luciferase（Fluc） were isolated from β-actin-Fluc transgenic mice and characterized by flow cytometry. Third passage AD-MSCs were divided into seven groups ： control ; hypoxia 6 hrs/reoxygen 2 hrs ; H/R ＋DMSO; H/R＋ LXR agonist T0901317（1μmol/L,5 vmol/L,10 μmol/L,15 μmol/L,respectively）. Cell survival conditions were assessed by bioluminesence imaging（BLI） via IVIS system. The protein expression of NF-κB was analyzed by Western blot. The secretion of TNF-α and IL-6 was tested by ELISA in each group. Results The AD-MSCs were both CD44＋ and CD90＋ assayed by flow cytometry. BLI proved that the firefly luciferase expression of AD-MSCs positively correlated with cell number （r^2 =0.97） ,and the pretreatment with LXR agonist could improve the survival of AD-MSCs from H/R injury（P〈0.05）. The levels of NF-κB,TNF-α and IL-6 in H/R group were much higher than those in control group（P〈0.01）. The levels of NF-κB,TNF-α and IL-6 in 10μmol/L LXR agonist group were lower than those in H/R group（P〈0.05,P〈0.01）. Conclusion LXR agonist has protective effects on the survival of AD-MSCs under H/R environment by suppressing NF-κB signaling.

关 键 词：间质干细胞 缺氧 荧光素酶类 萤火虫 细胞增殖 NF-κB 白细胞介素6 肿瘤坏死因子α 

分 类 号：R363[医药卫生—病理学]