The Effects Of Pg-LPS On Expression Of Endoplasmic Reticulum Stress Response In Adipocytes

Objective:Simulating the process of inflammation induced by periodontitis pathogenic factor in adipocytes.3T3-l1 preadipocytes were cultured in vitro and induced to differentiate into adipocytes,(Porphyromonas gingivalis lipopolysaccharide)Pg-LPS was used to effect on 3t3-l1 adipocytes.Objective to investigate the effect of Pg-LPS on the expression of GRP78,XBP1 s in the endoplasmic reticulum stress(ER),to investigate the effects of Pg-LPS on the endoplasmic reticulum stress state of adipose tissue.To investigate whether this effect is related to insulin resistance,and to further reveal the relationship between periodontitis and the development of T2 DM,To provide theoretical basis for clinical treatment and prevention.Methods:1?3T3-l1 preadipocytes were induced to differentiate into adipocytes,and cell differentiation was identified by oil red staining.2?The cells were divided into 2 group:blank control group,LPS treatment group.Blank control group :Add Serum free deme culture medium containing 0.8% bovine serum albumin,7 time points :0,30 min,1H,2h,4h,8h,12 h sampling;LPS treatment group:LPS treatment group was divided into 4 groups,respectively add 50ng/ml,100ng/ml,500ng/ml,1000ng/ml 4 concentration of Pg-LPS,Serum free deme culture medium containing 0.8% bovine serum albumin?7 time points 0,30 min,1H,2h,4h,8h,12 h sampling.To detect the expression level of the mRNA of GRP78 and XBP1 s which is the endoplasmic reticulum reaction marker.Results:1?After induction of differentiation of 14 d,3T3-l1 cells could be seen in lipid droplets under the microscope,3T3-l1 cells were differentiated into adipocytes?2?After Pg-LPS added for 12 hours,in the control group,there was no obvious cell apoptosis except for a few dead cells.Compared with the control group,in the experimental group,there was a small area of cell apoptosis and aggregation.3?The relative expression level of GRP78 in the control group was maintained at a relatively stable baseline from 0-12 H,Early intervention with Pg-LPS,The expression level of GRP78 in each concentration groups in the experimental group appears to be significantly higher and reached the peak value and then returned to the baseline level.50ng/ml concentration group,the expression level gradually increased after 30 minutes and reached the peak value at 1hour,drops to near baseline level at 2hour time point;100ng/ml concentration group,the relative expression of GRP78 mRNA gradually increased during 0-1hour,rise to peak in a high-speed during 1hour-2hour,and drops to near baseline level at 4 hour.500ng/ml concentration group: reached the peak value at 30 minutes,and droped to near baseline level at 2hour.1000ng/ml concentration group:the relative expression of GRP78 mRNA gradually increased during 0-1hour,reached the peak value at 1hour,and drops to near baseline level at 2 hour.4?The relative expression level of XBP1 s in the control group was maintained at a relatively stable baseline from 0-12hour;Early intervention with Pg-LPS,the expression level of XBP1 s in each concentration groups in the experimental group appears to be significantly higher and reached the peak value and then returned to the baseline level.50ng/ml concentration group:the expression level gradually increased after 30 minutes and reached the peak value at 1hour,drops to near baseline level at 2hour time point;100ng/ml concentration group,the relative expression of XBP1 s mRNA gradually increased during 0-1hour,rise to peak in a high-speed during 1hour-2hour,and drops to near baseline level at 4 hour.500ng/ml concentration group: reached the peak value at 30 minutes,and droped to near baseline level at 1hour.1000ng/ml concentration group:the relative expression of XBP1 s mRNA gradually increased during 0-1hour,reached the peak value at 1hour,and drops to near baseline level at 2 hour.Conclusion:1?Pg-LPS can increase the level of endoplasmic reticulum stress response in adipocytes,the expression of GRP78 and XBP1 s mRNA was significantly increased in adipocytes induced by Pg-LPS;2?There was no correlation between the expression level of GRP78,XBP1 s mRNA and the concentration and time of Pg-LPS;3?Pg-LPS may activate the apoptosis mechanism of adipocytes through endoplasmic reticulum stress.