The Effect Of Peripheral Serotonin On Osteoblasts Of The Isolated Sprague-Dawley Rats

Objective Objective to study the dose-related relationship between serotonin and osteoblasts(Osteoblasts,OBs)in SD rats.Methods(1)Isolate and culture the neonatal skull osteoblasts of the SD rats by adherent culture,and observe the cell morphology and determine the nature by using fluorescence stain.(2)Add serotonin intervention of different concentrations(10-5 mol/L?10-6 mol/L?10-7 mol/L?10-8 mol/L?10-9 mol/L?0mol/L)to cell culture fluid of the P3 generation.(3)On the fifth day,reflect the effect on the proliferation of different interventions concentration by MTT.(4)On the 10 th day,respectively,measure alkaline phosphatase(ALP)by using micro enzyme labeled methods and detect the type I collagen(Col-?)by immunofluorescence respectively,which was used to reflect the effect of different intervention concentration on the differentiation.(5)On the 15 th day,ELISA were used to measure the osteocalcin(OCN)level,which reflects the effect of different levels of intervention concentrations on mineralization.Statistical method To collect the data and analyze it by SPSS20.0 statistical software.Statistical data were showed by averages ± standard deviation,using single-factor variance to analyze the comparison of among the different groups,P <0.05 indicates that the difference has the statistical significance.Results the average optical density values of the intervention of serotonin of different concentrations(10-5 mol / L,10-6 mol / L,10-7 mol / L,10-8 mol / L,10-9 mol / L,0 mol / L)detected by MTT were 0.400 ± 0.028,0.379 ± 0.023,0.379 ± 0.047,0.401 ± 0.038,0.420 ± 0.038,0.618 ± 0.064,respectively.The difference has statistical significance(P<0.05).The average levels of ALP detected by micro enzyme labeled methods were 0.046 ± 0.002,0.017 ± 0.002,0.026 ± 0.002,0.046 ± 0.002,0.051 ± 0.002,0.073 ± 0.003.The difference between any group and blank control group has statistical significance(P<0.05).The average levels of Col-?detected by immunofluorescence method were 45.581 ± 6.378,33.110 ± 1.161,23.327 ± 0.784,15.731 ± 2.243,27.331 ± 2.430 and 33.592 ± 2.684 respectively,the difference has statistical significance(P <0.05)between any group and blank control group(P <0.05)except for 10-6 mol /l.The mean levels of OCN detected by ELISA were 3.002 ± 1.639,1.711 ± 0.370,2.193 ± 0.551,2.439 ± 1.469 and 2.703 ± 0.351 respectively.The difference has statistical significance(P <0.05)between any group and the blank control group.Conclusion Serotonin on osteoblasts into a negative effect,this negative effect and serotonin concentration was dose-dependent,with its concentration increased first strong and weak after the relationship.