Effect Of Conditioned Medium From TNF-? Pretreatment HUCMSC On Proliferation And Apoptosis Of HUVEC

Objective To investigate effect of conditioned medium(CM)from 100ng/mL TNF-? pretreatment human umbilical cord mesenchymal stem cells(hUCMSCs)on HUVEC injury and apoptosis induced by TNF-?.Methods HUVEC were randomly divided into 3 groups: control group,CM group and pretreatment-CM group.After 100 ng/m L TNF-? stimulating HUVEC 12 h,media in control groupwas replaced for the original medium,and conditioned medium and preconditioning medium were replaced for the CM and TNF-? pretreatment-CM,respectively.Cell proliferation was detected by MTT.Cell cycle was detected by flow cytometry.Cell apoptosis was assessed by AO-EB staining and flow cytometry.Statistic analyzed by t-test.Results At 24,48 and 72 h post treatment,the optical density values of control group were 0.51,043 and 0.32,and those in CM and pretreatment-CM groups were 0.56,0.51,0.45 ? 0.59,0.56,0.52,respectively.At the same time point post treatment,the proliferation numbers in CM and TNF-? pretreatment-CM groups were markedly higher than that in control group,as well as that in TNF-? pretreatment-CM group was the highest in three groups.Meanwhile,the result of cell cycle,which was assayed by flow cytometry,showed similar tendency with MTT result.At 24 h,48h and 72 h post treatment,the G2/M+S ratio of control group were(22.34±1.03),(10.66±1.07)and(9.58±0.82),and those in CM and TNF-? pretreatment-CM groups were(27.95±1.85),(18.59±1.48),(13.02±1.91)and(31.17±1.29),(22.44±2.28),(16.28±1.11),respectively.At the same time point post treatment,the G2/M+S ratio in CM and TNF-? pretreatment-CM groups were markedly higher than that in control group,as well as that in TNF-? pretreatment-CM group was the highest in three groups(F=58.793,P<0.05).Furthermore,the results of flow cytometry were showed that cell apoptosis ratesof control group were(18.95±1.01),(16.66±1.51)and(11.37±1.01),and those in CM and TNF-? pretreatment-CM groups were(12.04±1.13),(10.88±1.39),(6.88±1.63)and(8.16±1.44),(6.97±1.34),(2.97±1.44)at.24 h,48h and 72 h post treatment.At the same time point post treatment,the apoptosis ratios in CM and TNF-? pretreatment-CM groups were markedly lower than that in control group,as well as that in TNF-? pretreatment-CM group was the lowest in three groups(F=119.372,P<0.05).The results of flow cytometry were similar with those of AO-EB.Conclution The conditioned medium from 100ng/m L TNF-? pretreatment hUCMSCs significantly increased HUVEC proliferation activities and deceased cell apoptosis numbers and apoptosis rates,which were induced by TNF-?.