Lmwh Inhibits Tnf-Alpha And Il-6 In Placental Villous Explants And Its Effects Are Attenuated By Tlr-4/Nf-Kappa B P65 Blocking In Jeg-3 Cells

Preeclampsia (PE) leads to high perinatal mortality and morbidity due to hypertensive disorder and maternal inflammatory response of pregnancy. Studies demonstrated higher levels of TNF-alpha and IL-6 were observed in cases of PE. This research aims to explore the effects of low molecular weight heparin (LMWH) on TNF-alpha and IL-6 and its preliminary potential mechanism through TLR-4/NF-kappa B p65 pathway in placental villous explants and JEG-3 cells. Placental villous explants derived from patients with PE, spontaneous preterm labor and first trimester termination of pregnancy were cultured under normoxia and hypoxia conditions respectively. Then different concentrations of LMWH were added to explants in hypoxia group. In terms of gene target study, JEG-3 cells were transfected by SiRNA against TLR-4 and NF-kappa B p65 individually. Protein and mRNA levels of TNF-alpha and IL-6 were measured by western-blot and PCR. Immunohistochemistry staining was used to locate relative proteins in placental villous explants. Placental villous explants under hypoxia condition had similar upregulated levels of TNF-alpha and IL-6 with that in PE group, and this upregulation could be inhibited by LMWH with 2.5 mu g/ml as the most effective concentration. TLR-4 and NF-kappa B p65 were both higher expressed in PE group than those in control group. Blocking of TLR-4 or NF kappa B p65 in JEG-3 cells could both lead to attenuation of anti-inflammatory properties of LMWH. The results suggested LMWH could inhibit inflammatory response in placenta villous explants by inhibiting TNF-alpha and IL-6 and the effects were possibly via TLR-4/NF-kappa B p65 pathway.