黄芪甲苷孵育脂肪源性干细胞对顺铂诱导肾小管上皮细胞凋亡的影响

背景：干细胞移植用于治疗急性肾损伤的有效性已经被多个研究证实，但其对肾小管上皮细胞损伤的修复机制尚不明确。 目的：观察黄芪甲苷孵育后的脂肪源性干细胞对顺铂诱导的肾小管上皮细胞凋亡的保护作用及机制。 方法：实验分为4组。2.5 μmol/L顺铂诱导肾小管上皮细胞 24 h，建立肾小管细胞损伤模型(顺铂损伤组)；将脂肪源性干细胞与损伤肾小管上皮细胞共培养(脂肪源性干细胞+损伤肾小管上皮细胞组)；利用Transwell小室将20 mg/L黄芪甲苷孵育脂肪源性干细胞48 h后与损伤肾小管上皮细胞共培养(黄芪甲苷孵育脂肪源性干细胞+损伤肾小管上皮细胞组)；以正常肾小管上皮细胞做对照(正常对照组)。 结果与结论：与肾小管上皮细胞损伤组相比，AV/PI和TUNEL结果均显示脂肪源性干细胞+肾小管上皮细胞组和20 mg/L 黄芪甲苷脂肪源性干细胞+肾小管上皮细胞组肾小管上皮细胞发生凋亡的比例和数量明显减少；ELISA结果表明20 mg/L黄芪甲苷脂肪源性干细胞+肾小管上皮细胞组胰岛素样生长因子1分泌显著提高(P < 0.05)；Western blot进一步显示20 mg/L 黄芪甲苷脂肪源性干细胞+肾小管上皮细胞组caspase-3蛋白水平明显下降，而Bcl-2的表达量明显增加(P < 0.05)。表明黄芪甲苷孵育的人脂肪源性干细胞对顺铂诱导的肾小管上皮细胞凋亡具有抑制作用，从而有利于肾小管损伤的早期恢复，其保护机制可能与增加胰岛素样生长因子1分泌，抑制caspase-3表达、上调Bcl-2水平有关。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：

Effects of adipose-derived stem cells cultured with astragaloside on cisplatin-induced apoptosis of renal tubular epithelial cells

BACKGROUND:The validity of stem cell transplantation for acute kidney injury has been verified by many studies. However, the mechanism of repair of tubular epithelial cell injury remains unclear. OBJECTIVE:To investigate protective effects and mechanisms of adipose-derived stem cells cultured with astragaloside on the apoptosis of renal tubular epithelial cells induced by cisplatin. METHODS:There were four groups. Renal tubular epithelial cells were induced by 2.5 μmol/L cisplatin for 24 hours. Models of renal tubular cell injury were established in the cisplatin group. Adipose-derived stem cells were cocultured with renal tubular epithelial cells of injured kidney in the adipose-derived stem cells + renal tubular epithelial cells group. Using Transwell chamber, adipose-derived stem cells were incubated with 20 mg/L astragaloside for 48 hours, and then cocultured with renal tubular epithelial cells in the astragaloside-incubated adipose-derived stem cells + renal tubular epithelial cells group. Normal renal tubular epithelial cells served as  controls (normal control group). RESULTS AND CONCLUSION:Compared with renal tubular epithelial cell injury group, AV/PI and TUNEL results demonstrated that the apoptotic rate and number of renal tubular epithelial cells were apparently reduced in the adipose-derived stem cells + renal tubular epithelial cells group and 20 mg/L astragaloside-incubated adipose-derived stem cells + renal tubular epithelial cells group. ELISA results displayed that insulin-like growth factor-1 levels were significantly elevated in the 20 mg/L astragaloside-incubated adipose-derived stem cells + renal tubular epithelial cells group (P < 0.05). Western blot assay results exhibited that caspase-3 protein levels were noticeably diminished, but Bcl-2 expression was significantly increased in the 20 mg/L astragaloside-incubated adipose-derived stem cells + renal tubular epithelial cells group (P < 0.05). Results suggested that astragaloside-incubated human adipose-derived stem cells suppressed the apoptosis of renal tubular epithelial cells induced by cisplatin, which contributes to the early recovery of injured renal tubule. Its protective mechanism was probably associated with the secretion of insulin-like growth factor-1, inhibition of caspase-3 expression and upregulation of Bcl-2 levels.