Metformin Corrects Glucose Metabolism Reprogramming and NLRP3 Inflammasome-Induced Pyroptosis via Inhibiting the TLR4/NF-κB/PFKFB3 Signaling in Trophoblasts: Implication for a Potential Therapy of Preeclampsia

Figure 6

Pharmacological concentration of MET reduced the transcription of PFKFB3 via blocking transcription factor NF-κB1 directly binding on PFKFB3 promoter. HTR-8/SVneo cells were transfected with NF-κB1 plasmid for overexpression; 200 ng/ml LPS and 10 μM MET were subsequently incubated. (a) Representative images of NF-κB1 immunostaining in HTR-8/SVneo cells in different groups. At least 6 images per condition were analyzed. Scale bar: 50 μm. (b) Consensus binding motif of NF-κB1 from the JASPAR database and the NF-κB1 peak in the PFKFB3 promoter were found using the USCS database. (c) In the ChIP assay, PCR products of PFKFB3 in different groups are shown. DNA ChIP-ed with nonspecific IgG was used as a negative control. (d) Dual-luciferase reporter assays were performed to determine the PFKFB3 promoter activity in HTR8/SVneo cells in different groups. (e) qRT-PCR analysis of PFKFB3 mRNA levels in HTR-8/SVneo cells in different groups. Data are shown as the from three independent experiments. , , and by Student’s -test. SD: standard deviation; NF-κB1-OE: NF-κB1 overexpression plasmid.

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