Placental Tissues Of Preeclampsia Patients Research Articles

Protective role of complement factor H against the development of preeclampsia.

Pregnancy is an immunologically regulated, complex process. A tightly controlled complement system plays a crucial role in the successful establishment of pregnancy and parturition. Complement inhibitors at the feto-maternal interface are likely to prevent inappropriate complement activation to protect the fetus. In the present study, we aimed to understand the role of Factor H (FH), a negative regulator of complement activation, in normal pregnancy and in a model of pathological pregnancy, i.e. preeclampsia (PE). The distribution and expression of FH was investigated in placental tissues, various placental cells, and in the sera of healthy (CTRL) or PE pregnant women via immunohistochemistry, RT-qPCR, ELISA, and Western blot. Our results showed a differential expression of FH among the placental cell types, decidual stromal cells (DSCs), decidual endothelial cells (DECs), and extravillous trophoblasts (EVTs). Interestingly, FH was found to be considerably less expressed in the placental tissues of PE patients compared to normal placental tissue both at mRNA and protein levels. Similar results were obtained by measuring circulating FH levels in the sera of third trimester CTRL and PE mothers. Syncytiotrophoblast microvesicles, isolated from the placental tissues of PE and CTRL women, downregulated FH expression by DECs. The present study appears to suggest that FH is ubiquitously present in the normal placenta and plays a homeostatic role during pregnancy.

Preeclampsia-associated lncRNA FEZF1-AS1 regulates cell proliferation and apoptosis in placental trophoblast cells through the ELAVL1/NOC2L axis

BackgroundLncRNAs have been shown to be involved in and control the biological processes of multiple diseases, including preeclampsia (PE). The impairment of trophoblast cell proliferation is recognized as a significant anomaly contributing to the development of PE. LncRNA FEZF1-AS1 was found downregulated in placental tissues of PE patients. However, the precise regulatory mechanism of FEZF1-AS1 in placental trophoblast proliferation and apoptosis remains unclear.ResultsIn this study, we conducted an investigation into the expression levels of FEZF1-AS1 and NOC2L in placental tissues obtained from patients diagnosed with PE. Subsequently, we employed CCK-8 and EdU assays to quantify cell proliferation, while TUNEL staining and western blot for apoptosis-related protein detection to assess apoptosis. Furthermore, the interactions between FEZF1-AS1 and ELAVL1, as well as NOC2L and ELAVL1, were confirmed through the implementation of RIP and RNA pull-down assays. We found a downregulation of lncRNA FEZF1-AS1 and NOC2L in placental tissues of PE patients. Overexpression of FEZF1-AS1 or NOC2L resulted in increased cell proliferation and inhibition of apoptosis, whereas knockdown of FEZF1-AS1 or NOC2L had the opposite effect. In addition, lncRNA FEZF1-AS1 stabilized NOC2L mRNA expression by interacting with ELAVL1. Moreover, partial reversal of the effects of FEZF1-AS1 overexpression on cell proliferation and apoptosis was observed upon suppression of ELAVL1 or NOC2L.ConclusionsPE related lncRNA FEZF1-AS1 could regulate apoptosis and proliferation of placental trophoblast cells through the ELAVL1/NOC2L axis.

Upregulation of SEMP1 Contributes to Improving the Biological Functions of Trophoblast via the PI3K/AKT Pathway in Preeclampsia.

Disturbance of extravillous trophoblast infiltration is associated with preeclampsia (PE), a severe condition of pregnancy characterized by hypertension and proteinuria. Senescence-associated epithelial membrane protein 1 (SEMP1), an integral membrane protein, is a vital component of tight junction strands in epithelial or endothelial cells, with no clear function reported in PE. Gene Expression Omnibus (GEO) datasets showed that SEMP1 expression was downregulated in the placental tissues of PE patients, which was confirmed by assessing SEMP1 levels in placental samples collected in our hospital. Furthermore, less SEMP1 was detected in cytokeratin 7 positive trophoblast cells in the spiral arteries of rat placentas post L-arginine methyl ester hydrochloride (L-NAME) treatment. Trophoblast cells acquired robust ability of proliferation, migration, and invasion when SEMP1 was overexpressed. Such capability was weakened in SEMP1-silenced cells. Trophoblast cells overexpressing SEMP1 secreted more vascular endothelial growth factor A (VEGFA), which facilitated the tube formation of human umbilical vein endothelial cells. Blockade of PI3K/AKT signaling transduction with LY294002 dampened the effects of SEMP1 on trophoblast cells. Collectively, we firstly indicated that SEMP1 inhibition is a potential driver for PE, which may be associated with the deactivation of the PI3K/AKT pathway.

Long non-coding RNA TLR8-AS1 induces preeclampsia through increasing TLR8/STAT1 axis.

Our current study tried to assay the role of long noncoding RNAs (lncRNAs) TLR8-AS1 in regulating preeclampsia. TLR8-AS1 expression was examined in the clinical placental tissues of preeclampsia patients and the trophoblast cells induced by lipopolysaccharide (LPS). Then, different lentivirus was infected into trophoblast cells to study the role of TLR8-AS1 in cell functions. Furthermore, interactions among TLR8-AS1, signal transducer and activator of transcription 1 (STAT1) and toll-like receptor 8 (TLR8) were determined. A rat model of preeclampsia induced by N(omega)-nitro-L-arginine methyl ester was developed to validate the in-vitro findings. High expression of TLR8-AS1 was detected in placental tissues of preeclampsia patients and LPS-induced trophoblast cells. In addition, overexpression of TLR8-AS1 arrested the proliferation, migration and invasion of trophoblast cells, which was related to the upregulation of TLR8 expression. Mechanistically, TLR8-AS1 recruited STAT1 to bind to the TLR8 promoter region, and thus promoted the transcription of TLR8. Meanwhile, overexpression of TLR8-AS1 was shown to aggravate preeclampsia by elevating TLR8 in vivo . Our study confirmed that TLR8-AS1 aggravated the progression of preeclampsia through increasing the expression of STAT1 and TLR8.

The endocannabinoidome in human placenta: Possible contribution to the pathogenesis of preeclampsia.

Preeclampsia (PE) was first reported thousands of years ago, yet there is still a shortage of biomarkers to determine the severity and type of PE. The importance of the expanded endocannabinoid system, or endocannabinoidome (eCBome), has emerged recently in placental physiology and pathology, though the potential alterations of the eCBome in PE have not been fully explored. Analysis by qRT-PCR using placental samples of normotensive and PE women demonstrate for the first time the presence of ABHD4, GDE1, and DAGLβ in both normotensive and PE placental tissues. Interestingly, NAPE-PLD, FAAH-1, DAGLα, MAGL, and ABHD6 mRNA levels were increased in the placental tissues of PE patients. Quantification in plasma and placental tissues showed a decrease for anandamide (AEA), N-oleoylethanolamine (OEA), and N-docosahexaenoylethanolamine (DHEA) in the placenta, accompanied only by a decrease in plasma levels of AEA. In addition, a strong negative correlation was obtained between OEA and the biomarker of PE, soluble fms-like tyrosine kinase-1. Given the inflammatory nature of PE and the anti-inflammatory role of OEA and DHEA, the decrease in the local levels of these mediators may underlie the inflammatory component of this pathology. Additionally, lower AEA levels in both placenta and plasma may contribute to the atypical alterations of the spiral arteries in PE due to the vasorelaxation effects of AEA. These results add new information to the role of the eCBome members in placental development, while also pointing to a potential role as biomarkers of PE.

Integrated Proteomic and N-Glycoproteomic Profiling of Placental Tissues of Patients with Preeclampsia.

Preeclampsia (PE) is a multi-system disorder of pregnancy that poses a serious threat to maternal and perinatal health worldwide. This study aims to evaluate the global alterations of protein expression and N-glycosylations that are crucial for PE pathogenesis. Here, tandem mass tag labeling combined with LC-MS/MS was employed to determine the global expression of all proteins and intact glycopeptide in placentas from three healthy pregnant women, three patients with early-onset severe PE, and three patients with late-onset severe PE. A total of 2260 proteins were quantified across 9 placental tissues, of which 37 and 23 were differentially expressed in the early-onset and late-onset PE groups, compared to the controls. A total of 789 glycopeptides were accurately quantified, which were derived from 204 glycosylated sites in 159 glycoproteins and were modified by 59 N-Linked glycans. A total of 123 differently expressed glycopeptides, which were from 47 glycoproteins were identified among three groups. Through a combined analysis of proteomic and glycoproteomic data, it was found that the changes in 10 glycoproteins were caused by the difference in glycosylation level but not in the protein abundance level. This is the first study to conduct an integrated proteomic and glycoproteomic characterization of placental tissues of PE patients. Our findings suggest that glycosylation modification may affect the molecular function of proteins through changes in the glycosylation structure or the occupancy of glycosylation, which will provide new insights to help elucidating the pathogenic mechanism of PE.

Downregulation of LINC00221 promotes angiopoiesis in HUVEC and inhibits recruitment of macrophages by augmenting miR-542-3p in trophoblast cells.

To investigate the effects of long intergenic non-protein coding RNA 221 (LINC00221) on preeclampsia (PE) and its mechanism. The expression of LINC00221 was detected in placental tissues from PE patients and normal pregnant women (non-PE). Next, the effects of LINC00221 silencing on trophoblast cells (HTR-8/SVneo and JEG-3) and co-cultured HUVECs or macrophages were evaluated. Afterwards, miR-542-3p was confirmed to bind to LINC00221 directly, and miR-542-3p mimics and inhibitors were transfected into trophoblast cells. Next, a rescue experiment was performed to examine the effect of LINC00221/miR-542-3p axis. Finally, the effect of LINC00221 was also verified in vivo in rat PE models. The expression of LINC00221 was higher in placental tissues of PE patients than those of non-PE. LINC00221 silencing significantly reduced MCP1 level and increased the VEGF level in trophoblast cells. LINC00221 knockdown in trophoblast cells remarkably enhanced VEGFR expression and the angiopoiesis of HUVECs, and decreased the migration and invasion of macrophages and reduced TNF-α level. Besides, LINC00221 knockdown decreased CHOP, p-IREα, p-PERK, and iNOS expression and increased Trx expression. Notably, LINC00221 negatively regulated miR-542-3p expression. MiR-542-3p overexpression had an effect to that of LINC00221 knockdown, while miR-542-3p inhibition had the opposite effect. Treatment with miR-542-3p inhibitors partially reversed the protective effect of LINC00221 silencing. PE rat model results were consistent with those of in vitro experiments. Downregulation of LINC00221 might reduce dysfunction, inflammatory responses, endoplasmic reticulum stress, and oxidative stress, and thereby protect against PE by augmenting miR-542-3p.

HDAC5 inactivates CYR61-regulated CD31/mTOR axis to prevent the occurrence of preeclampsia.

Our study was to pinpoint the significance of histone deacetylase 5 (HDAC5) affecting the pathogenesis of preeclampsia (PE) via CD31/mammalian target of rapamycin (mTOR) axis by regulating cysteine-rich angiogenic inducer 61 (CYR61). Expression of HDAC5, CYR61, and CD31/mTOR in placental tissues of patients with PE and trophoblast cells HTR-8/SVneo cells was determined first followed by their interaction analysis. Following different transfection, the significance of HDAC5 in cell functions was assayed in relation to CYR61 and CD31/mTOR. An in vivo PE mouse model was constructed for further validation. The clinical tissue and in vitro cell experimentations discovered that HDAC5 was downregulated in placental tissues of PE patients and trophoblast cells, while CYR61, CD31, mTOR, and p-mTOR displayed upregulation. After overexpression of HDAC5, trophoblast cell functions were enhanced. HDAC5 reduced the acetylation enrichment of H3K27 to inhibit the expression of CYR61. Furthermore, CYR61 promoted the activation of CD31/mTOR axis, thereby inhibiting HTR-8/SVneo cell functions. The in vivo rat model confirmed the above alterations. Taken together, HDAC5 contributes to downregulation of CYR61 through histone deacetylation, inactivating CD31/mTOR axis, which prevents the occurrence and development of PE.

MiR-200b-3p is upregulated in the placental tissues from patients with preeclampsia and promotes the development of preeclampsia via targeting profilin 2

ABSTRACT Preeclampsia is a serious pregnancy disorder affecting both maternal and fetal health. However, the pathogenesis of preeclampsia has not been fully understood. This study aimed to investigate the key microRNAs (miRNAs) in the development of preeclampsia. A high-throughput miRNA sequencing analysis for the placental tissues from patients with preeclampsia and healthy controls was conducted, followed by investigation of differentially expressed miRNAs (DEMs) and functional enrichment analysis. Moreover, the expression of a key DEM, named miR-200b-3p, in the preeclampsia patients was validated, and the effects of miR-200b-3p overexpression on the proliferation, migration, and apoptosis of HTR8 trophoblast cells were investigated in vitro. Furthermore, the target gene of miR-200b-3p was investigated based on gene expression profile GSE177049 and miRWalk 2.0 database. The target relationship between miR-200b-3p and profilin 2 (PFN2) was investigated in vitro. A total of 12 DEMs including miR-200b-3p were identified between preeclampsia placental tissues and control placental tissues, which were significantly enriched in several pathways, such as cell adhesion molecules (CAMs) and tight junction. Moreover, increased expression of miR-200b-3p was revealed in the placental tissues of preeclampsia patients, and overexpression of miR-200b-3p suppressed cell proliferation and migration but promoted apoptosis of trophoblast cells. Furthermore, PFN2 was confirmed as a target of miR-200b-3p, and overexpression of PFN2 reversed the inhibitory effects of miR-200b-3p overexpression on trophoblast cell migration. Our findings reveal that miR-200b-3p is upregulated in the placental tissues of patients with preeclampsia and promotes preeclampsia development via PFN2. miR-200b-3p may serve as a promising therapeutic target against preeclampsia.

MicroRNA-140-5p from human umbilical cord mesenchymal stem cells–released exosomes suppresses preeclampsia development

This work unraveled the action of human umbilical cord mesenchymal stem cells-released exosomes (huc-MSCs-EXO) transfer of miR-140-5p in preeclampsia (PE). miR-140-5p and follistatin-like 3 (FSTL3) expression in placental tissues of PE patients was tested. EXO were isolated from huc-MSCs. Hypoxic trophoblast cells were co-cultured with huc-MSCs-EXO. Cell biological functions, angiogenesis, and inflammation were evaluated. Suppressed miR-140-5p and induced FSTL3 levels were measured in PE. Huc-MSCs-EXO drove biological functions and angiogenesis while hindering inflammation in hypoxic trophoblast cells. Increasing miR-140-5p further improved the positive role of huc-MSCs-EXO for hypoxic trophoblast cells, but the miR-140-5p-mediated effect in hypoxic trophoblast cells was abrogated by overexpressing FSTL3. miR-140-5p from huc-MSCs-EXO suppresses PE through repressing FSTL3.

Circ_0085296 inhibits the biological functions of trophoblast cells to promote the progression of preeclampsia via the miR-942-5p/THBS2 network.

Insufficient invasion of trophoblast cells is one of the important causes of preeclampsia (PE). Circular RNA (circRNA) has been proven to regulate the biological functions of trophoblast cells and mediate the progression of PE. The expression of circ_0085296, microRNA (miR)-942-5p, and thrombospondin 2 (THBS2) was detected by quantitative real-time PCR. In addition, the interaction between miR-942-5p and circ_0085296 or THBS2 was confirmed by dual-luciferase reporter assay and RIP assay. Our data showed that circ_0085296 was upregulated in the placental tissues of PE patients. Silenced circ_0085296 could enhance the proliferation, migration, invasion, and angiogenesis of HTR-8/SVneo cells. Besides, circ_0085296 was found to act as miR-942-5p sponge. Function analysis results suggested that miR-942-5p inhibitor reversed the positive regulation of circ_0085296 knockdown on the biological functions of HTR-8/SVneo cells. Moreover, THBS2 was a target of miR-942-5p, and its overexpression also reversed the promotion effect of miR-942-5p on the proliferation, migration, invasion, and angiogenesis of HTR-8/SVneo cells. Also, circ_0085296 was discovered to positively regulate THBS2 by sponging miR-942-5p. To sum up, our results revealed that circ_0085296 could inhibit trophoblast cells proliferation, migration, invasion, and angiogenesis by regulating miR-942-5p/THBS2, confirming that circ_0085296 might be a potential therapeutic target for PE.

The Mechanism of Downregulation of Twist1 Inhibiting Trophoblast Invasion and Aggravating the Development of Preeclampsia.

To study the expression of under expressed transcription factor Twist1 in preeclampsia (PE) and its effect on the invasion of placental trophoblast cells and to explore its related mechanism on the development of PE by establishing a pregnant rat model. Methods: the villi were collected from the induced abortion in the first trimester (6–8 weeks), the normal placenta (18–20 weeks) induced by the second trimester, the term placenta tissue of normal pregnancy (37–40 weeks), and the placental tissue of patients with PE, to detect the expression of Twist1. Trophoblast cells were subjected to primary culture in placental tissues of normal pregnant women and placental tissues of PE patients. The invasion ability of the two groups of trophoblasts was detected, and the primary cultured trophoblasts were divided into two groups: an experimental group and a control group. Specific Twist1 siRNA was added to the experimental group, and no reagents were added to the control group. The above-mentioned cells were given different interventions. To explore the effect of Twist1 on trophoblast cell invasion, cells were cultivated for 72 h. The SD rats were conceived. After the pregnancy was stable, the SD rats in different groups were treated with different treatments (interference with Twist1), and the average systolic blood pressure and urine protein of the gestational mothers in the different treatment groups were measured at 1 week, 2 weeks, and full-term pregnancy. The expression of Twist1 in the placenta tissue of SD rats with different interventions at full-term pregnancy was detected. The results showed that Twist1 expression is down-regulated in PE, and the invasion ability of placental trophoblast cells in PE patients is weak. After inhibiting Twist1, the mean tail artery pressure and urine protein level of SD pregnant rats increase, showing a trend of PE. The mechanism may be related to the inhibition of the placenta by Twist1 Trophoblast cell invasion.

EID1 plays a protective role in early-onset pre-eclampsia via promoting proliferation and invasion in trophoblast cells

Pre-eclampsia is a pregnancy-specific syndrome, which is partly due to abnormal proliferation and invasion of trophoblast cells. EP300 interacting inhibitor of differentiation 1 (EID1) participates in cell proliferation and invasion. This study aims to investigate the roles of EID1 in trophoblast cells and pre-eclampsia. The expression of EID1 in placental tissues from 60 women with pre-eclampsia and 60 health pregnancies was detected by real-time PCR and immunohistochemical staining. EID1 was overexpressed or silenced by transfection of plasmid or siRNA in HTR-8/SVneo trophoblast cells, and then cell proliferation, cell cycle transition, migration, and invasion were determined by CCK-8 assay, flow cytometry, immunofluorescent staining, immunoblotting, and transwell assays. In addition, the activity of Akt/b-catenin signaling was measured by immunofluorescent staining and Western blot. EID1 mRNA level was decreased in placental tissues of pre-eclampsia patients, especially early-onset pre-eclampsia, accompanied by more severe clinical manifestation and a higher rate of fetal growth restriction (FGR). Gain- and loss-of-function experiments demonstrated that EID1 promoted proliferation and cell cycle transition, migration, and invasion in HTR-8/SVneo cells and its knockdown played opposite roles, suggesting that EID1 may be required for normal gestation. Akt/b-catenin signaling was activated after EID1 forced expression and deactivated after its silencing. EID1 promoted proliferation and invasion of cultured trophoblast cells with possible involvement of Akt/b-catenin signaling. These findings may provide novel insights for the diagnosis and treatment of early-onset pre-eclampsia in a clinic.

Melatonin regulates trophoblast pyroptosis, invasion and migration in preeclampsia by inhibiting HtrA1 transcription through the microRNA-520c-3p/SETD7 axis.

Melatonin has an inhibitory effect on preeclampsia (PE). This study was launched to explore the way that melatonin regulated trophoblast migration, invasion, and pyroptosis in PE and to provide new ideas for the diagnosis and treatment of PE. Expression levels of melatonin receptors (MT1 and MT2), microRNA (miR)-520c-3p, SETD7, and HtrA1 in placental tissues and HTR8/SVneo cells were measured by RT-qPCR and Western blot. Scratch, Transwell, and Western blot assays were performed to detect migration, invasion, and pyroptosis of hypoxia/reoxygenation (H/R)-treated HTR8/SVneo cells. Dual-luciferase reporter assay was utilized to verify the targeting relationship between miR-520c-3p and SETD7. ChIP experiment was conducted to detect the enrichment of H3K4me3 and SETD7 in HtrA1 promoter. Low expression of MT1, MT2, and miR-520c-3p and high expression of SETD7 and HtrA1 were observed in the placental tissues of PE patients and H/R-treated HTR8/Svneo cells. A high concentration of melatonin promoted migration and invasion and inhibited pyroptosis of PE cell models. Knockdown of miR-520c-3p, overexpression of SETD7, or overexpression of HtrA1 impaired migration and invasion and accelerated pyroptosis of H/R-treated HTR8/SVneo cells, but these outcomes could be reversed by treatment with 1000 μM melatonin. miR-520c-3p targeted SETD7 which promoted histone methylation in the promoter region of HtrA1. Melatonin may inhibit HtrA1 transcription through the miR-520c-3p/SETD7 axis to promote trophoblast invasion and migration and reduce trophoblast pyroptosis in PE.

CircPAPPA Regulates the Proliferation, Migration, Invasion, Apoptosis, and Cell Cycle of Trophoblast Cells Through the miR-3127-5p/HOXA7 Axis.

Abnormal function of trophoblast cells is one of the important causes of preeclampsia (PE). Circular RNA (circRNA) is thought to be involved in the regulation of various diseases progression, including PE. However, the role of circRNA pregnancy-associated plasma protein A (circPAPPA) in PE is less studied. The expression levels of circPAPPA, miR-3127-5p, and homeobox A7 (HOXA7) were determined by quantitative real-time PCR. Cell proliferation was evaluated using MTT assay and colony formation assay. Besides, flow cytometry was used to detect cell apoptosis and cell cycle distribution. In addition, the interaction between miR-3127-5p and circPAPPA or HOXA7 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. CircPAPPA was lowly expressed in the placental tissues of PE patients. Knockdown of circPAPPA inhibited proliferation, migration, and invasion, while induced apoptosis and cell cycle arrest in trophoblast cells. MiR-3127-5p could be targeted by circPAPPA, and its inhibitor reversed the effect of circPAPPA silencing on the biological function of trophoblast cells. Moreover, HOXA7 was a target of miR-3127-5p. HOXA7 overexpression reversed the effect of miR-3127-5p on the biological function of trophoblast cells. Our research indicated that circPAPPA positively regulated the biological function of trophoblast cells to mediate the progression of PE by miR-3127-5p/HOXA7 axis, which suggested that circPAPPA might be a potential biomarker for PE.

MiR-133b regulates oxidative stress injury of trophoblasts in preeclampsia by mediating the JAK2/STAT3 signaling pathway.

Preeclampsia (PE) is a pregnancy-related syndrome. Aberrant placental microRNAs (miRNAs) expression might associate with PE, including miR-133b. However, its role in the pathogenesis of preeclampsia remains elusive. Therefore, this study explored the role of miR-133b in oxidative stress injury of trophoblasts in preeclampsia (PE) by mediating the JAK2/STAT3 signaling pathway. Placental tissues were collected from PE patients to detect the expression of miR-133b and JAK2/STAT3. Then, in vitro experiments were performed on human extravillous trophoblast-derived HTR-8/SVneo cells, which were divided into Normal, hypoxia/reoxygenation (H/R), H/R + miR-NC, H/R + miR-133b inhibitor, H/R + JAK2 siRNA and H/R + miR-133b inhibitor + JAK2 siRNA groups. Cell invasion and migration abilities were detected by Transwell and wound healing assays, while apoptosis was detected by flow cytometry. The intracellular oxidative stress levels were also measured. Furthermore, the expression of miR-133b and the JAK2/STAT3 pathway was determined by qRT-PCR and Western blotting. We found that miR-133b was up-regulated, with decreases in JAK2 and p-STAT3/STAT3 in placental tissues of PE patients. Additionally, HTR8/SVneo cells in the H/R group had decreased invasion and migration abilities with increased apoptotic rates and oxidative stress levels. Moreover, the expression of miR-133b was up-regulated with decreases in p-JAK2 and p-STAT3 in H/R-treated HTR8/SVneo cells. These indicators in the H/R + miR-133b inhibitor group were ameliorated in comparison with those in the H/R group but deteriorated in the H/R + JAK2 siRNA group. Moreover, JAK2 siRNA reversed the positive effect of the miR-133b inhibitor on the invasion and migration abilities of trophoblasts. In summary, inhibiting miR-133b may improve oxidative stress injury to promote the migration and invasion of trophoblasts and suppress apoptosis by activating the JAK2/STAT3 pathway.

P–422 Acylglycerol Kinase is a new novel marker in preeclampsia

Abstract Study question What is the potential function of Acylglycerol Kinase (AGK) in the pathogenesis of preeclampsia(PE). Summary answer AGK plays an important role in the pathogenesis of PE by influencing the function of the trophoblast cells. What is known already PE is the leading cause of maternal and perinatal mortality and morbidity. The underlying mechanism is still not completely elucidated. Disorder migration, invasion and mitochondria function of trophoblast cells are one of mechanism in preeclampsia. AGK is a subunit of the mitochondrial channel protein complex TIM22, and maintain the stability of mitochondrial structure and function. Studies have shown that AGK is related to the development of various cancers by infecting the cells migration and invasion. It will be interesting to explore the potential function of AGK in the process of early trophoblast development and the pathogenesis of PE. Study design, size, duration Firstly, explore the expression of AGK in PE. Secondly, lentivirus systems was used to generate loss and gain of function models in trophoblast cell line-HTR8/Sneov to study the role of AGK in the pathogenesis of PE. Participants/materials, setting, methods We examined the expression of AGK both in placental tissues from PE patients and normal pregnant patients. Meanwhile, we generated AGK loss and gain of function models in HTR8/Sneov cells by using lentivirus systems. Transwell assays, scratch-wound assays, EDU and plate clone formation assays, cell apoptosis assays, cell cycle assays, ATP concentration, mtDNA level and transmission electron microscopy were used to examine the function of AGK in HTR8/Sneov cells model. Main results and the role of chance In this study, AGK was significantly decreased in the extra-villous trophoblast (EVT) cells in placental tissues of PE patients compared with that in control group by immunohistochemistry. And further confirmed the expression of AGK in placental tissues by QPCR, western blot. We demonstrated that knockdown of AGK in HTR8 dramatically decreased the cell proliferation, migration and invasion. It also showed the significantly lower plate clone formation rate in AGK knockdown -HTR8 cells compare with the WT HTR8 cells. While overexpression of AGK in HTR8 dramatically increased the cell proliferation, migration, invasion and higher plate clone formation rate. Further, we demonstrated that AGK regulated the ATP level and mtDNA level in HTR8 cells model. And we found that knockdown AGK decreased the number of mitochondria, and shown the mitochondrial crista disorder, mitochondrial swelling and dissolution and mitochondrial membrane fragmentation. While overexpression AGK increased the number of mitochondria. Limitations, reasons for caution Although we show that AGK played an important role in the function of HTR8, the study of working mechanism of AGK in PE is still very limited. More studies will be performed to explore its underline mechanism. Wider implications of the findings: This study was the first time to explore the role of AGK in PE. It will help us to better understand the pathogenesis of PE, which might be helpful in future application of novel therapeutic targets in PE. Trial registration number Not applicable

Circ_0011460 upregulates HTRA1 expression by sponging miR‐762 to suppress HTR8/SVneo cell growth, migration, and invasion

Aberrant expression of circular RNAs (circRNAs) during placental development could affect fetal growth and contribute to preeclampsia (PE). Circ_0011460 was found to be differentially expressed in placental samples of PE. However, the exact function and mechanism of circ_0011460 in PE process remain largely undefined. Levels of circ_0011460, microRNA (miR)-762, and high-temperature requirement-A serine peptidase 1 (HTRA1) were detected using quantitative real-time polymerase chain reaction and Western blot. In vitro experiments in HTR8/SVneo cells were conducted using cell counting kit-8, wound healing, transwell, flow cytometry and Western blot assays. The direct interactions between miR-762 and circ_0011460 or HTRA1 were verified using dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays. Circ_0011460 possessed a loop structure and was highly expressed in placental tissues of PE patients. Overexpression of circ_0011460 greatly suppressed HTR8/SVneo cell proliferation, migration, and invasion, and accelerated cell apoptosis. While circ_0011460 knockdown yielded the opposite trends on above biological behaviors. Mechanistically, we confirmed that circ_0011460 could up-regulate HTRA1 expression via serving as a sponge of miR-762. Further rescue studies demonstrated that circ_0011460 exerted its roles via targeting miR-762, and miR-762 promoted HTR8/SVneo cell growth, migration and invasion via regulating HTRA1. In all, circ_0011460 suppressed HTR8/SVneo cell growth, migration, and invasion via miR-762/HTRA1 axis, suggesting a new insight into the pathogenesis of PE.

Hsa_circ_0001326 regulates proliferation, migration, invasion, and EMT of HTR-8/SVneo cells via increasing IL16 expression.

Preeclampsia (PE) is a serious pregnancy complication. It has been shown that insufficient infiltration of extravillous trophoblasts (EVTs) is related to the pathogenesis of PE. Circular hsa_circ_0001326 (circ_0001326) has been uncovered to be upregulated in PE. However, the influence of circ_0001326 on the infiltration of trophoblasts is indistinct. 48 pregnant women (25 PE patients and 23 healthy controls) were recruited for this study. Human trophoblasts HTR-8/Svneo were used for function analysis. Expression of circ_0001326 was evaluated by real-time quantitative polymerase chain reaction (RT-qPCR). The diagnostic value of circ_0001326 was analyzed by receiver operating characteristic (ROC) curve. Cell proliferation, migration, and invasion of HTR-8/Svneo cells were determined using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), wound-healing, and transwell assays. Protein levels were detected using Western blotting. The regulation mechanism of circ_0001326 was surveyed by bioinformatics analysis and dual-luciferase reporter assay. Circ_0001326 was highly expressed in plasma samples and placental tissues of PE patients. Moreover, plasma circ_0001326 could distinguish PE patients from healthy controls (area under the curve (AUC) =0.793). Functionally, circ_0001326 overexpression repressed HTR-8/Svneo cell proliferation, epithelial-mesenchymal transition (EMT), migration, and invasion, but circ_0001326 silencing had the opposing impact. Mechanically, circ_0001326 regulated interleukin-16 (IL16) expression via sponging microRNA (miR)-558. MiR-558 mimic or IL16knockdown overturned the inhibiting effect of circ_0001326 overexpression on HTR-8/Svneo cell proliferation, EMT, migration, and invasion. Circ_0001326 elevated IL16 expression via sponging miR-558, thus curbing HTR-8/Svneo cell proliferation, EMT, migration, and invasion, suggesting that circ_0001326 might be involved in the pathogenesis of PE.

Extracellular vesicle-derived microRNA-18b ameliorates preeclampsia by enhancing trophoblast proliferation and migration via Notch2/TIM3/mTORC1 axis.

Preeclampsia (PE), a common disorder of pregnancy, is characterized by insufficient trophoblast migration and inadequate vascular remodelling, such that promotion of trophoblast proliferation might ameliorate PE. In the current study, we sought to study the underlying mechanism of extracellular vesicle (EV)‐derived microRNA‐18 (miR‐18b) in PE. Human umbilical cord mesenchymal stem cells (HUCMSCs) isolated from placental tissues were verified through osteogenic, adipogenic and chondrogenic differentiation assays. Bioinformatics analyses and dual‐luciferase reporter gene assay were adopted to confirm the targeting relationship between miR‐18b and Notch2. The functional roles of EV‐derived miR‐18b and Notch2 in trophoblasts were determined using loss‐ and gain‐of‐function experiments, and trophoblast proliferation and migration were assayed using CCK‐8 and Transwell tests. In vivo experiments were conducted to determine the effect of EV‐derived miR‐18b, Notch2 and TIM3/mTORC1 in a rat model of PE, with monitoring of blood pressure and urine proteinuria. TUNEL staining was conducted to observe the cell apoptosis of placental tissues of PE rats. We found down‐regulated miR‐18b expression, and elevated Notch2, TIM3 and mTORC1 levels in the placental tissues of PE patients compared with normal placenta. miR‐18b was delivered to trophoblasts and targeted Notch2 and negatively its expression, whereas Notch2 positively mediated the expression of TIM3/mTORC1. EV‐derived miR‐18b or Notch2 down‐regulation enhanced trophoblast proliferation and migration in vitro and decreased blood pressure and 24 hours urinary protein in PE rats by deactivating the TIM3/mTORC1 axis in vivo. In summary, EV‐derived miR‐18b promoted trophoblast proliferation and migration via down‐regulation of Notch2‐dependent TIM3/mTORC1.