Inconformity of CXCL3 Plasma Level and Placenta Expression in Preeclampsia and Its Effect on Trophoblast Viability and Invasion

<div><p>As a member of the chemokine family, CXCL3 was previously known to participate in many pathophysiological events. However, whether CXCL3 stimulates trophoblast invasion as a key process of preeclampsia pathogenesis remains largely unknown. Therefore, the aim of this study was to investigate this hypothesis and determine the effect of CXCL3 on the first trimester trophoblast. Seventy gravidas were included in this study. ELISA was used to detect CXCL3 plasma levels on preeclampsia and normal pregnant groups. CXCL3 protein and mRNA levels were detected via Western blot and real-time quantitative PCR analysis after immunolocalized in human placenta. Moreover, the CXCL3 function in HTR-8/Svneo was analyzed via WST-1 assay, flow cytometry and invasion test. The plasma CXCL3 level in preeclampsia was significantly higher than that in normal pregnancy. CXCL3 expression was observed in the cytoplasm of placental trophoblasts and vascular endothelium in all groups without significant difference between maternal and fetal sides. In addition, placenta CXCL3 expression in severe preeclampsia was significantly lower than those in normal and mild PE groups. Moreover, exogenous CXCL3 can promote the proliferation and invasion of HTR-8/Svneo; however, its effect on apoptosis remains unclear. In summary, a significant abnormality of plasma CXCL3 level and placental CXCL3 expression was discovered in severe preeclampsia; CXCL3 had a function in trophoblast invasion, which indicated its participation in shallow implantation. Therefore CXCL3 might be involved in severe preeclampsia pathogenesis.</p></div

Plasma CXCL3 concentration.

<p>A:Plasma CXCL3 levels of normal pregnant women, mild preeclamptic pregnant women and severe preeclamptic pregnant women. Middle line: median; Whisker: standard deviation. B: The correlation between plasma CXCL3 levels of the severe PE group participants with 24-hour urinary protein.</p

A: Invasion trophoblast cells stained by eosin under microscope (×400).

<p>A: 0 ng/ml rhCXCL3; B: 1 ng/ml rhCXCL3; C: 10 ng/ml rhCXCL3; D: 50 ng/ml rhCXCL3; E: 100 ng/ml rhCXCL3; F: 200 ng/ml rhCXCL3. G: Effect of rhCXCL3 on the invasiveness of trophoblast cells. Bar: mean; Whisker: standard deviation. a: p<0.05, versus 0 ng/ml; b: p<0.05, versus 1 ng/ml.</p

Clinical characteristics of study participants.

<p>Data are presented as mean ± standard error of the mean.</p><p>☆:Full term pregnancy compared with preterm pregnancy in mild PE group, P<0.05;</p><p>△:Severe PE group compared with normal pregnant group, P<0.05;</p><p>▴:Severe PE group compared with mild PE group, P<0.05.</p><p>Clinical characteristics of study participants.</p

Effect of rhCXCL3 on the viability of trophoblast cells.

<p>Bar: mean; Whisker: standard deviation. A: HTR-8/SVneo cell viability in different rhCXCL3 concentration. B: Cell viability at different time intervals (12, 24, 48, and 72 h) . a: p<0.05, versus 12 h; b: p<0.05, versus 24 h. C: Cell viability in different rhCXCL3 concentration. a: a: p<0.05, versus 0 ng/ml; b: p<0.05, versus 1 ng/ml; c: p<0.05, versus 10 ng/ml; d: p<0.05, versus 50 ng/ml.</p

CXCL3 expression in placentae of three groups (Immunohistochemistry, SP, ×400).

<p>(S: syncytiotrophoblast; V: vascular endothelial cells) A: CXCL3 expressed on maternal surface of normal placentae; B: CXCL3 expressed on maternal surface of mild preeclamptic placentae; C: CXCL3 expressed on maternal surface of severe preeclamptic placentae; D: CXCL3 expressed on fetal surface of normal placentae; E: CXCL3 expressed on fetal surface of mild preeclamptic placentae; F: CXCL3 expressed on fetal surface of severe preeclamptic placentae; G: negative control; H: human colon adenocarcinoma tissue as positive control.</p

Additional file 1 of Cerebral venous congestion alters CNS homeostatic plasticity, evoking tinnitus-like behavior

Supplementary Material

Cell Membrane-Anchored DNA Nanoinhibitor for Inhibition of Receptor Tyrosine Kinase Signaling Pathways via Steric Hindrance and Lysosome-Induced Protein Degradation

Receptor tyrosine kinase (RTK) plays a crucial role in cancer progression, and it has been identified as a key drug target for cancer targeted therapy. Although traditional RTK-targeting drugs are effective, there are some limitations that potentially hinder the further development of RTK-targeting drugs. Therefore, it is urgently needed to develop novel, simple, and general RTK-targeting inhibitors with a new mechanism of action for cancer targeted therapy. Here, a cell membrane-anchored RTK-targeting DNA nanoinhibitor is developed to inhibit RTK function. By using a DNA tetrahedron as a framework, RTK-specific aptamers as the recognition elements, and cholesterol as anchoring molecules, this DNA nanoinhibitor could rapidly anchor on the cell membrane and specifically bind to RTK. Compared with traditional RTK-targeting inhibitors, this DNA nanoinhibitor does not need to bind at a limited domain on RTK, which increases the possibilities of developing RTK inhibitors. With the cellular-mesenchymal to epithelial transition factor (c-Met) as a target RTK, the DNA nanoinhibitor can not only induce steric hindrance effects to inhibit c-Met activation but also reduce the c-Met level via lysosome-mediated protein degradation and thus inhibition of c-Met signaling pathways and related cell behaviors. Moreover, the DNA nanoinhibitor is feasible for other RTKs by just replacing aptamers. This work may provide a novel, simple, and general RTK-targeting nanoinhibitor and possess great value in RTK-targeted cancer therapy

Expressions of OPG protein in different groups (Gray Value).

<p>Expressions of OPG protein in different groups (Gray Value).</p

The OPG mRNA level in human placental tissues.

<p>The OPG mRNA level in severe preeclamptic group was much higher than those of the mild preeclamptic and normal control group.</p