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  

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

作　　者：王葳[1] 姜燕[1] 王巍巍[1] 张金元[1] 

机构地区：[1]解放军第四五五医院肾脏科,南京军区肾脏病研究所,上海市200052

出　　处：《中国组织工程研究》2014年第28期4498-4503,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81100493);上海市基础研究重大项目子课题(12DJ1400203);上海市优秀青年医学人才培养计划(XYQ2011012);南京军区医学科技创新项目重点课题(10Z008)~~

摘　　要：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。2.5μmol/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。BACKGROUND：The validity of stem celltransplantation for acute kidney injury has been verified by many studies. However, the mechanism of repair of tubular epithelial cellinjury 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 2.5μmol/L cisplatin for 24 hours. Models of renal tubular cellinjury 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 Transwel 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 cellinjury 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 supp

关 键 词：干细胞 脂肪干细胞 黄芪甲苷 人脂肪源性干细胞 急性肾损伤 肾小管上皮细胞 顺铂 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]