Abstract: Objective To study the autophagy of rat adipose derived stem cells ( ADSC) and effects of oxygen glucose deprivation on autophagy and lipogenesis of rat ADSC. Method The ADSC were prepared conventionally and set up as the control group ( group G) , the low glucose group ( group SG) , the hypoxia group ( group G + CoCl 2 ) , the hypoxia & low glucose group ( group SG + CoCl2 ) and the autophagy positive control group ( group EBSS) . In different culture periods, cell viability was measured by MTT assays, the autophagy by Western blot and adipogenic induction, the oil red O staining and its OD value were performed. Result The ADSC in all groups were cultured and survived for 24 hours without difference ( P>0. 05) in different culture periods. But at 48 and 72 hours, the cell proliferation of the group G was significantly higher than that of the other 3 groups( P<0. 05) . By Western blot, the protein expression of LC3-Ⅰ was significantly less than that of LC3-Ⅱ in groups G,SG,SG+CoCl2 and EBSS. However, the LC3-Ⅰ s in groups G+CoCl2 and SG+CoCl2 were much higher than that in the group EBSS. In the test of adipogenesis, the OD value of the group G was the highest, which was statistically different ( P < 0. 05 ) , compared with other groups. Conclusion The rat ADSC may have a high level of autophagy. It was suggested that under the circumstance of sugar deficiency and hypoxia, inhibiting autophagy will hinder the survival and lipogenesis of rat ADSC.

Key words: rat adipose derived stem cells, autophagy, sugar deficiency, hypoxia

摘要： 目的 研究大鼠脂肪干细胞( adipose derived stem cells,ADSC)及糖氧剥夺下,细胞自噬和对诱导分化成脂的影响。 方法常规制备 ADSC, 设 置 对 照 组 ( G 组 ) 、 少 糖 组 ( SG 组 ) 、 缺 氧 组 ( G + CoCl2 组 ) 、 缺氧少糖组( SG+CoCl2 组)和自噬阳性对照组( EBSS 组) 。 设置不同培养时间,MTT 检测各组 ADSC 存活情况;Western blot 分析细胞自噬及成脂诱导油红“ O”染色与比色。 结果 不同培养时间,ADSC 细胞均可增殖。 培养 24 h,各组细胞之间无差别( P>0. 05) ,但培养 48 h 和 72 h,G 组细胞增殖明显高于其它三组( SG 组、G + CoCl2 组和 SG+CoCl2 组) ,P<0.05;Western blot 分 析, EBSS 组、G 组 和 SG 组 的自噬相关蛋白 Ⅰ 型 ( LC3-Ⅰ ) 明显弱于自噬相关蛋白Ⅱ 型( LC3-Ⅱ ) ,且 G+CoCl2 组和 SG+CoCl2 组的 LC3-Ⅰ 明显高于 EBSS 组;成脂诱导实验中,G 组油红“ O” 比色值最高,与其它组比较均有统计学差异( P<0. 05) 。 结论 ADSC 具有较强的细胞自噬作用,推测在缺糖缺氧条件下,通过抑制细胞自噬,降低 ADSC 存活率以及诱导分化成脂率。

关键词: 大鼠脂肪干细胞, 细胞自噬, 缺糖, 缺氧