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. Author manuscript; available in PMC: 2011 Jan 26.
Published in final edited form as: Circulation. 2010 Jan 11;121(3):436–444. doi: 10.1161/CIRCULATIONAHA.109.902890

Figure 4. TNF-α is the serum factor in autoantibody-injected pregnant mice that is responsible for induction of sEng production by endothelial cells.

Figure 4

(A) HUVEC cells were treated with Ang II, IgG from preeclamptic or normotensive women and the concentration of sEng in cell culture supernatant was determined. Data are expressed as mean (± SEM) of ≥3 experiments performed in duplicate (n=12–14 patient’s IgG for each group). (B) HUVEC cells treated with serum from control pregnant mice, pregnant mice injected with IgG from normotensive (NT) or preeclamptic women (PE) or coinjected with preeclamptic IgG and losartan (PE+Losartan), 7-aa epitope peptide (PE+7-aa) at day 5 following the initial injections. HUEVC cells also treated with anti-TNF-α, Enbrel or hemin for 30 minitues before adding serum from pregnant mouse injected with IgG from women with preeclampsia (mouse serum (PE)). After 24 hour the sEng concentration in cell culture supernatants were measured. * P < 0.05 versus cells treated with serum from mouse injected with normotensive IgG. ** P < 0.05 versus cells treated with serum from mouse injected with preeclamptic IgG. Data are expressed as mean ± SEM. N =8 mice’s serum for each group. (C) HUVEC cells were treated with TNF-α in the presence of TNF-α neutralizing antibody, Enbrel or hemin for 24 hours. Concentration of sEng in cell culture media was determined by ELISA. * P < 0.01 versus control untreated cells. ** P < 0.05 versus cells treated TNF-α alone. Data are expressed as mean (± SEM) of ≥3 experiments performed in duplicate (n=9 for each group). (D) IgG from normotensive or preeclamptic pregnant women were introduced into pregnant mice at gestation days 13 and 14. Mouse serum was collected at gestation days 18 and levels of TNF-α were measured by ELISA. Data are expressed as mean ± SEM, n=5–8 for each group. * P < 0.05 versus mice treated with normotensive IgG. ** P < 0.05 versus mice treated with preeclamptic IgG alone. (E) Placentas were collected (n=8–10 for each group) at gestation day 18 (5 days following the initial IgG injection). Real-time PCR was performed to quantify the TNF-α mRNA abundance. Data are expressed as mean ± SEM. * P < 0.001 versus normotensive IgG injection. ** P < 0.01 versus preeclamptic IgG injection..