Effect Of PTH_1-34 Reverse The Inhibit Of AGEs On Osteogenic Differentiation Of BMSCs

Objective:To investigate whether parathyroid hormone(PTH_1-34)can antagonize the effect of advanced glycosylation end products（AGEs）that inhibit osteogenic differentiation of bone marrow mesenchymal stem cell（BMSCs）by decreasing expression of transforming growth factor-βand selerstin in steocytes.Methods:1.BMSCs which were taked from femur and tibia of 4 weeks SD rats were isolated and cultured in vitro.2.Randomly divided into control group（BSA）、 experimental group,AGEs with 4concentration gradients（25ug/mL,50ug/mL,100ug/mL,200ug/mL）act on BMSCs72 hours,Cell proliferation was detected using MTT after being isolated and cultured,Then we choose 200ug/mL AGEs and the corresponding concentration of bovine serum albumin（BSA）act on BMSCs,measured 2-7 days cell proliferation.The mRNA expression of sost and TGF-βwas detected using real time quantitative after cultured in vitro 7days,Alizarin red staining was used to compare the status of mineralized nodule formation after being cultured in vitro 21 days.3.200ug/ml AGEs were added for 48 h after pretreatments of 10-8M PTH_1-34 for6 h,repeat above experience（LSD-t Test）.Results:1.Rat bone marrow mesenchymal stem cells at the third passage werefusiform,spindle-shaped and had high refraction.2.Treatment of the BMSCs with AGEs alone could inhibited cell proliferation（P <0.05）,PTH_1-34 could reversed the effect of advanced glycosylation end products（AGEs）that inhibit proliferative effect of bone marrow mesenchymal stem cell（BMSCs）.3.Treatment of the BMSCs with AGEs with 7 day significantly increased the expression of TGF-β/sostmRNA（P<0.05）,however,pretreatment of the cells with PTH_1-34 could inhibit the expression of TGF-β /sostmRNA.There were significant differences in expression rates of SOST/TGF-βmRNA（F=23.98,P=0.00;F=17.61,P=0.00）between AGEs group and PTH+AGEsgroup.4.Alizarin red staining shows that AGEs decress the status of mineralized nodule formation,intermittent treat the cell with PTH could partly reserve this detrimental effects.Conclution:Parathyroid hormone(PTH_1-34)can antagonize the effect of advanced glycosylation end products（AGEs）that inhibit osteogenic differentiation of bone marrow mesenchymal stem cell（BMSCs）by decreasing expression of SOST、TGF-βmRNA.