Objective:To establish the aging model of NCTC clone 929（L929）induced by advanced glycation end products（AGEs）and to explore its biological mechanism.Methods:In this study

L929 was randomly divided into five groups :blank control group（cultivated L929 with common incubation media）

experimental group A（L929+AGEs

0.05 g/L）

experimental group B（L929+AGEs

0.1 g/L）

experimental group C（L929+AGEs

0.2 g/L）

positive control group（L929+D-galactose

10 g/L）.Cell proliferation was detected by CCK-8 at 24

48

72 hours.The establishment of aging model in L929 was evaluated by senescence aging β-galactosidase（SA-β-gal）staining and cell cycle assay

and the expressions of p16 and CyclinD1 were detected by Western blot.The levels of reactive oxygen species associated with oxidative stress in the cells

and aging indicators such as superoxide dismutase（SOD）

malondialdehyde（MDA）and catalase（CAT）were examined.Results:CCK8 showed that AGEs could inhibit L929 proliferation and cell viability was decreased significantly after being cultivated for 48 hours.Compared with blank control group

AGEs increased the activity of β-galactosidase（B:24.0±3.4

C:43.0±10.0

P =0.000）and the blocking rate of phase G1（A:69.90±2.13

B:74.74±1.49

C:79.29±3.38

P=0.000）and protein expression of p16（B:0.53±0.20

C:0.63±0.11

P=0.001

P=0.000） in a dose-dependent manner

while AGEs decreased CyclinD1 expression[（1.32±0.09） vs.（0.51±0.16）

P=0.000].AGEs decreased the activity of SOD and CAT（B:29.19±6.67

C:15.13±5.21

P =0.001

P =0.000;A:3.73±0.42

B:1.59±0.16

C:1.20±0.20

P=0.000）and increased the level of MDA and reactive oxygen species [（1.19±0.08）vs.（2.39±0.07）

P=0.000;（1.47±0.04）vs.（1.84±0.09）

P=0.020].The number of cells in phase G1 and β-galactosidase positive staining cells was significantly lower in experimental group A than in positive control group [（12.0±3.6）vs.（45.0±8.2）

P=0.004;（69.90±2.13）vs.（76.59±0.61）

P=0.001].The expression of p16 protein was significantly decreased in experimental group A than in positive control group[（0.34±0.12）vs.（0.64±0.04）

P=0.001]

while the expression of CyclinD1 protein was higher than that in positive control group[（1.32±0.09）vs.（0.51±0.16）

P=0.000].The activities of SOD and CAT were significantly higher than those of positive control group[（39.53±4.24）vs.（27.09±6.25）

P=0.018;（3.73±0.42）vs.（1.64±0.09）

P=0.000]

while the levels of MDA and reactive oxygen species were significantly lower than those in positive control group[（1.19±0.08）vs.（2.39±0.07）

P=0.000;（1.47±0.04）vs.（1.84±0.09）

P=0.02].There was no significant difference between experimental B and C and positive control group in these respects（P>0.05）.Conclusion:0.1 g/L and 0.2 g/L AGEs can establish aging L929 in vitro model

which may be related to the changes of oxidative stress and senescence-associated protein expression of p16 and CyclinD1.