糖氧剥夺对大鼠脂肪干细胞自噬和成脂的影响  

Effects of Oxygen Glucose Deprivation on Autophagy and Lipogenesis of Rat Adipose Stem Cells

作　　者：袁志坚[1] 吴小瑜[1] 何文涓 丁亚明[1] 钟晓春 YUAN Zhijian;WU Xiaoyu;HE Wenjuan;DING Yaming;ZHONG Xiaochun(Wuxi Higher Health Vocational Technology School,Wuxi 214111,China;Medical School of Hangzhou Normal University,Hangzhou 311121,China)

机构地区：[1]无锡卫生高等职业技术学校,无锡214028 [2]杭州师范大学医学院,杭州311121

出　　处：《实验动物科学》2021年第6期11-15,共5页Laboratory Animal Science

基　　金：江苏省卫生计生委科研课题,No.J201616。

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+CoCl;组)、缺氧少糖组(SG+CoCl;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+CoCl;组和SG+CoCl;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+CoCl;组和SG+CoCl;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。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;),the hypoxia & low glucose group( group SG+CoCl;) 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+CoCl;and EBSS. However,the LC3-Ⅰs in groups G+CoCl;and SG+CoCl;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.

关 键 词：大鼠脂肪干细胞 细胞自噬 缺糖 缺氧 

分 类 号：Q952[生物学—动物学]