Different Times Of High Glucose On The Expression Of Visfatin And Collagen TypeⅠin Neonatal Rat Cardiac Fibroblast

Objective:This study is to clear whether the expression of visfatin andcollagen type I in cardiac fibroblasts by observing high glucose stimulaion, atdifferent times,on the expression of visfatin and collagen type I in vitrocultured neonatal rat cardiac fibroblasts;Then, explore the role of visfatin andcollagen typeⅠin the formation of myocardial fibrosis,and provide new ideasfor diagnosis,prevention and treatment of diabetes-induced cardiovasculardisease.Method:Primary cultures cardiac fibroblas from the hearts of2-3days’neonatal Sprague-Dawley(SD) rats aseptically,regardless of sex.Then,weshould add high concentration of glucose DMEM medium containing10%fetal bovine serum,and culture myocardial fibroblasts in5%CO2incubators at37°C for2-3days. Next,passage these cells at1:2,when cells grow to theintegrated state.And this experiment uses2-3generation cells.After48hours’culture with serum and24hours’culture without serum,cells in goodconditions are selected and put into6groups randomly.①low concentration ofglucose control group (the NG group,5.5mmol/L,).Different time of highconcentration of glucose stimulation groups(the HG groups,30mmol/L),5groups:②6hours group,③12hours group,④24hours group,⑤48hoursgroup,⑥the hypertonic group (the OC group). After the same concentration ofhigh glucose (30mmol/L) for different time,use GAPDH as the internalcontrol,then detect the expression of mRNA in visfatin and procollagen typeⅠwith real time-PCR and the expression of protein of visfatin and procollagentypeⅠin medium with ELISA.Results:1the effect of different times’ high glucose (30mmol/L) on the expression ofvisfatin-mRNA in cardiac fibroblas:12hours group,24hours group,and48hours group,the expression of visfatin-mRNA,are all higher than theexpression of low concentration of glucose control group’,among the groups,the differences are statistically significant(P<0.05).the highestexpression of visfatin-mRNA is24hours group’;in12hours group,24hoursgroup,and48hours group,the expression of visfatin-mRNA is increasedgradually,and the difference between each group is statistically significant(P<0.05); the expression of visfatin-mRNA in48hours group is slightly below24hours group, but no statistically significant (P>0.05); the expression ofvisfatin-mRNA in6hours group and the hypertonic group are slightly higherthan low concentration of glucose control group,but no statistically significant(P>0.05).2the effect of different times’ high glucose (30mmol/L) on the expression ofprocollagen typeⅠ’mRNA in cardiac fibroblas:12hours group,24hoursgroup,and48hours group,the expression of procollagen typeⅠ’ mRNA,are allhigher than the expression of low concentration of glucose controlgroup’,among the groups,the differences are statisticallysignificant(P<0.05).the highest expression of procollagen typeⅠ’ mRNA is24hours group’;in12hours group,24hours group,and48hours group,theexpression of procollagen type Ⅰ’mRNA is increased gradually,and thedifference between each group is statistically significant (P<0.05); theexpression of procollagen typeⅠ’ mRNA in48hours group is slightly below24hours group, but no statistically significant (P>0.05); the expressions ofprocollagen typeⅠ’ mRNA in6hours group and the hypertonic group areslightly higher than low concentration of glucose control group,but nostatistically significant (P>0.05).3the effect of different times’ high glucose (30mmol/L) on the expression ofvisfatin-protein in medium:12hours group,24hours group,and48hoursgroup,the expression of visfatin-protein in medium,are all higher than theexpression of low concentration of glucose control group’,among thegroups,the differences are statistically significant(P<0.05).the highestexpression of visfatin-protein is24hours group’;in12hours group,24hoursgroup,and48hours group,the expression of visfatin-protein is increasedgradually,and the difference between each group is statistically significant (P<0.05); the expression of visfatin-protein in48hours group is slightlybelow24hours group, but no statistically significant (P>0.05); the expressionsof visfatin-protein in6hours group and the hypertonic group are slightlyhigher than low concentration of glucose control group,but no statisticallysignificant (P>0.05).4the effect of different times’ high glucose (30mmol/L) on the expression ofcollagen typeⅠ’ protein in medium:12hours group,24hours group,and48hours group,the expression of collagen typeⅠ’ protein are all higher than theexpression of low concentration of glucose control group’,among thegroups,the differences are statistically significant(P<0.05).the highestexpression of collagen typeⅠ’ protein in medium is24hours group’;in12hours group,24hours group,and48hours group,the expression of collagentypeⅠ’mRNA is increased gradually,and the difference between each group isstatistically significant (P<0.05); the expression of collagen typeⅠ’ protein in48hours group is slightly below24hours group, but no statisticallysignificant(P>0.05); the expression of collagen typeⅠ’protein in6hoursgroup and the hypertonic group is slightly higher than low concentration ofglucose control group,but no statistically significant (P>0.05).Conclusion: The cardiac fibroblasts could express visfatin and collagentypeⅠ.Within24hours, high concentrations of glucose could promote theexpression of visfatin and collagen typeⅠin vitro cultured cardiac fibroblasts.And with the time extended gradually, the expression levels of mRNA andprotein in visfatin and collagen type I are increased significantly. High glucosefor the expressions of visfatin and collagen type Ⅰin cardiac fibroblasts aredue to the glucose concentration, not the osmotic pressure caused.