Effects Of Different FimA Variants Of P.gingivalis-rFimA On The Function Of Human Umbilical Artery Smooth Muscle Cells

Objective: ?n order to explore the effects of P.gingivalis–rFimA possessing ?,? fimA genotypegenotype Porphyromonas gingivalis on the function of human umbilical artery smooth muscle cells,including ability of proliferation,migration and phenotype transformation and the effects on the development of atherosclerosis.Methods: 1.?n vitro culture HUASMCs were seeded into 96-well plates using CCK-8 to detect different concentrations of P.gingivalis-rFimA(0.5 g/ml,1 g/ml,2 g/ml,5 g/ml,10g/ml)in 2h,8h,24 h,48h on the proliferation of smooth muscle cells,where the measured OD value at 450nm;2.HUASMCs were stimulated by 10?g/ml P.gingivalis-rFimA in subsequent experiment.Transwell migration assay and Ultrastructure was detected by transmission electron microscopy.The levels of Cell Retinol-Binding Protein-1 and smooth muscle Myosin heavy chain were determined by flow cytometry(FCM).Results: 1.At different time points,? type P.gingivalis-rFimA and ? type P.gingivalis-rFimA group HUASMCs proliferation effect than the negative control group,the difference was statistically significant(P<0.05).With the increase of the concentration of proliferation of HUASMCs ? type P.gingivalis-rFimA and ? type P.gingivalis-rFimA group with enhanced(P<0.05).At the same concentration and function time points,? type fimA type P.gingivalis-rFimA stimulated proliferation of HUASMCs was stronger than ? fimA P.gingivalis-rFimA(P<0.05).At different time points,? P.gingivalis-rFimA and HUASMCs ? type P.gingivalis-rFimA group migration cell number were higher than the negative control group,the difference was statistically significant(P<0.05).2h,? P.gingivalis-rFimA HUASMCs migration cell number than ? type P.gingivalis-rFimA group,but the difference was not statistically significant(P<0.05);in 8h,24 h and 48 h,? P.gingivalis-rFimA HUASMCs the number of the migrated cells than ? P.gingivalis-rFimA group,the differences were statistically significant(P<0.05).With the extension of time,the negative control group,? P.gingivalis-rFimA and HUASMCs ? P.gingivalis-rFimA group the number of the migrated cells were increased(P<0.05).HUASMCs were stimulated by ?,? fimA genotype P.gingivalis-rFimA.After 2h,8h,24 h,48h,the HUASMCs were observed by transmission electron microscope(TEM).By electron microscopy,?,? P.gingivalis-rFimA 2H HUASMCs stimulation group,obvious rough endoplasmic reticulum,mitochondria,glycogen 8h,increased phagosomes and lysosomes,organelles have different degrees of swelling;type ? in P.gingivalis-rFimA stimulation group at each observation point were observed in the lipid droplets and increased;? type fimA type P.gingivalis-rFimA stimulation group in 24 h and 48 h,cells begin to dissolve,and most of the cell structure damage.4.At different time points,? P.gingivalis-rFimA and ? P.gingivalis-rFimA after stimulation were observed in the synthesis of type HUASMCs up-regulated the expression of CRBP-1 molecular markers,marker expression contraction type molecule SM-MHC,? and P.gingivalis-rFimA were upregulated the expression of CRBP-1 was stronger than in the ? group P.gingivalis-rFimA,24 h and 48 h in ? P.gingivalis-rFimA strong expression of SM-MHC was down regulated in ? P.gingivalis-rFimA group(P < 0.05)Conclusion:? type,? type P.gingivalis-rFimA can stimulate the smooth muscle cell phenotype from contractile phenotype into the more activity synthetic phenotype.and promote the proliferation and migration,suggesting that ?t is suggested that infection and invasion of type ?,type ? P.gingivalis-rFimA may induce the proliferation and migration of smooth muscle cells,thereby forming the components of atherosclerotic plaques,?n addition,the infection and invasion of type ? and type ? P.gingivalis-rFimA may lead to the dysfunction of smooth muscle cells and play a role in the occurrence and development of AS and different P.gingivalis-rFimA on smooth muscle cell proliferation,there are some differences between the effects of migration and phenotype conversion.