摘要
为了探讨胰岛素样生长因子1(IGF-1)通过PI3K/Akt通路对大鼠软骨细胞衰老的影响,试验选取21日龄SD大鼠肱骨和股骨头上的组织碎片进行软骨细胞的分离培养,将第3代软骨细胞分为5组,即对照组、IGF-1组、抑制剂MK+IGF-1组、抑制剂MK组和DMSO组。加药处理在37℃、体积分数为5%CO 2条件下培养24 h后,进行SA-β-Gal染色;RT-PCR检测各组细胞p53、p21的mRNA水平;Western blot检测各组细胞Akt、p-Akt、p53、p21的蛋白水平。结果显示,IGF-1组的β-Gal染色阳性率、p53和p21的mRNA表达水平均极显著高于对照组(P<0.01);IGF-1组Akt、p-Akt、p53、p21的蛋白表达水平显著高于对照组(P<0.05);而抑制剂MK+IGF-1组与IGF-1组相比,β-Gal染色阳性率显著降低(P<0.05),相关分子mRNA和蛋白水平均显著下降(P<0.05)。以上结果表明,IGF-1可经PI3K/Akt信号通路来促进大鼠软骨细胞衰老。
In order to explore the effect of insulin-like growth factor-1(IGF-1)on chondrocytes senescence in rats through PI3K/Akt pathway,the tissue fragments of the tibia and femoral head of 21-day-old SD rats were removed for chondrocyte isolation and culture.The experiment was divided into five groups:control,treatment of IGF-1,IGF-1 addition to inhibitor MK,inhibitor MK and DMSO.The SA-β-Gal activity assay was estimated after 24 hours of drug treatment.The levels of p53 and p21 mRNA were determined by RT-qPCR.The expressions of Akt,p-Akt,p53 and p21 protein levels were detected by Western blot.The results showed that compared with control,the positive rates ofβ-Gal of IGF-1 group signifcantly increased(P<0.01).The mRNA levels of p53,p21 and the protein expression levels of Akt,p-Akt,p53 and p21 in IGF-1 group were signifcantly higher than that in control(P<0.05).Compared with IGF-1 group,the positive rate ofβ-Gal of inhibitor MK+IGF-1 group signifcantly declined(P<0.05),and the mRNA and protein expression levels of relevant molecules were markely decreased(P<0.05).In conclusion,IGF-1 can promote chondrocyte senescence in rats through PI3K/Akt pathway.
作者
赵丽东
丛淋淋
徐瑶
王颢然
吴俊楠
肖建华
ZHAO Li-dong;CONG Lin-lin;XU Yao;WANG Hao-ran;WU Jun-nan;XIAO Jian-hua(College of Veterinary Medicine,Northeast Agricultural University,Harbin,Heilongjiang,150030,China)
出处
《动物医学进展》
北大核心
2019年第10期64-68,共5页
Progress In Veterinary Medicine
