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In vitro effects of sEng and TGF-β on human umbilical vein endothelial cells and trophoblasts.
AIM: The aim of this study was to evaluate the in vitro effects of sEng and TGF-β on human umbilical vein endothelial cells (HUVECs) and trophoblasts.
METHODS: HUVECs and trophoblasts were treated with 1, 5, 10, 15, 30, 50, 80 and 100 μg/L sEng, TGF-β or (sEng + TGF-β) for 6, 12, 24, 36, 48 and 72 h, respectively. sEng, TGF-β and NO levels in culture media of HUVECs were measured by ELISA. Survival rates of HUVECs and trophoblasts were detected by 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT) assay. Apoptotic rate and cell cycle distribution were detected by flow cytometry. eNOS, MMP-2 and MMP-9 expressions were detected.
RESULTS: The survival rate of 100 μg/L of sEng-treated HUVECs was significantly lower than those of other groups at the same time point (P < 0.05). The survival rate of the TGF-β group was significantly lower than that of the sEng group after treatment at each concentration for 36 and 48 h (P < 0.05). NO content significantly decreased with increasing sEng concentration (r = 0.89, P < 0.05) and with extended time at identical sEng concentration (r = 0.78, P < 0.05). The number of membrane-penetrating trophoblasts in the control group significantly surpassed that of sEng group (P < 0.05). The number of expressions in the TGF-β group significantly exceeded that of control group (P < 0.05), being significantly different from that of sEng group (P < 0.05). MMP-2 and MMP-9 expressions in sEng group were significantly lower than those of the control group (P < 0.05), and such expressions of the TGF-β group significantly surpassed those of other groups (P < 0.05).
CONCLUSION: sEng and TGF-β were closely associated with endothelial cell injury and abnormalities of trophoblast proliferation and infiltration in vitro.
METHODS: HUVECs and trophoblasts were treated with 1, 5, 10, 15, 30, 50, 80 and 100 μg/L sEng, TGF-β or (sEng + TGF-β) for 6, 12, 24, 36, 48 and 72 h, respectively. sEng, TGF-β and NO levels in culture media of HUVECs were measured by ELISA. Survival rates of HUVECs and trophoblasts were detected by 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT) assay. Apoptotic rate and cell cycle distribution were detected by flow cytometry. eNOS, MMP-2 and MMP-9 expressions were detected.
RESULTS: The survival rate of 100 μg/L of sEng-treated HUVECs was significantly lower than those of other groups at the same time point (P < 0.05). The survival rate of the TGF-β group was significantly lower than that of the sEng group after treatment at each concentration for 36 and 48 h (P < 0.05). NO content significantly decreased with increasing sEng concentration (r = 0.89, P < 0.05) and with extended time at identical sEng concentration (r = 0.78, P < 0.05). The number of membrane-penetrating trophoblasts in the control group significantly surpassed that of sEng group (P < 0.05). The number of expressions in the TGF-β group significantly exceeded that of control group (P < 0.05), being significantly different from that of sEng group (P < 0.05). MMP-2 and MMP-9 expressions in sEng group were significantly lower than those of the control group (P < 0.05), and such expressions of the TGF-β group significantly surpassed those of other groups (P < 0.05).
CONCLUSION: sEng and TGF-β were closely associated with endothelial cell injury and abnormalities of trophoblast proliferation and infiltration in vitro.
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