Down-regulated placental miR-21 contributes to preeclampsia through targeting RASA1.

Human placenta was obtained from early onset preeclampsia, late onset preeclampsia, and their gestational age-matched normal pregnancy. Using RT-qPCR, western blot, and immunohistochemistry, it was demonstrated that miR-21 expressions were significantly decreased in preeclampsia while RASA1 were increased. Suppression of miR-21 in placental HTR-8/SVneo cells, remarkably upregulated RASA1, decreased proliferation, inhibited invasion, and promoted apoptosis of trophoblast cells, while overexpression of miR-21 alleviated these effects. Dual-luciferase reporter assays revealed RASA1 to be a direct target of miR-21 in trophoblast cells. miR-21 may serve key roles in the development of preeclampsia by targeting RASA1.

Identification of Diagnostic CpG Signatures in Patients with Gestational Diabetes Mellitus via Epigenome-Wide Association Study Integrated with Machine Learning.

BACKGROUND: Gestational diabetes mellitus (GDM) is the most prevalent metabolic disease during pregnancy, but the diagnosis is controversial and lagging partly due to the lack of useful biomarkers. CpG methylation is involved in the development of GDM. However, the specific CpG methylation sites serving as diagnostic biomarkers of GDM remain unclear. Here, we aimed to explore CpG signatures and establish the predicting model for the GDM diagnosis. METHODS: DNA methylation data of GSE88929 and GSE102177 were obtained from the GEO database, followed by the epigenome-wide association study (EWAS). GO and KEGG pathway analyses were performed by using the clusterProfiler package of R. The PPI network was constructed in the STRING database and Cytoscape software. The SVM model was established, in which the ß-values of selected CpG sites were the predictor variable and the occurrence of GDM was the outcome variable. RESULTS: We identified 62 significant CpG methylation sites in the GDM samples compared with the control samples. GO and KEGG analyses based on the 62 CpG sites demonstrated that several essential cellular processes and signaling pathways were enriched in the system. A total of 12 hub genes related to the identified CpG sites were found in the PPI network. The SVM model based on the selected CpGs within the promoter region, including cg00922748, cg05216211, cg05376185, cg06617468, cg17097119, and cg22385669, was established, and the AUC values of the training set and testing set in the model were 0.8138 and 0.7576. The AUC value of the independent validation set of GSE102177 was 0.6667. CONCLUSION: We identified potential diagnostic CpG signatures by EWAS integrated with the SVM model. The SVM model based on the identified 6 CpG sites reliably predicted the GDM occurrence, contributing to the diagnosis of GDM. Our finding provides new insights into the cross-application of EWAS and machine learning in GDM investigation.

Naringenin prevents pregnancy-induced hypertension via suppression of JAK/STAT3 signalling pathway in mice.

BACKGROUND: Pregnancy-induced hypertension (PIH) is characterized by high blood pressure during pregnancy, which causes perinatal and maternal mortality. Inflammation, oxidative stress and the JAK2/STAT3 signalling pathway have been reported to play critical roles in the pathogenies of PIH. Due to the safety and side effects of current treatments for PIH, searching for new therapeutic agents is urgently needed. Naringenin is a flavonoid with anti-inflammation and anti-oxidation activities. In the current study, the effects of naringenin on PIH were investigated. METHODS: We established the PIH mouse model and administrated naringenin to these mice. The blood pressure, total urine protein, plasma levels of vasodilation converting enzyme (VCE), α-1A adrenergic receptor (α-ADR) and angiotensin, inflammatory cytokines, oxidative stress markers were measured. The protein levels of reactive oxygen species proto-oncogene 1 (ROS1), superoxide dismutase 2 (SOD2), signal transducer and activator of transcription 3 (STAT3), phospho-STAT3, Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP-1), Janus kinase 2 (JAK2) and phospho-JAK2, in vascular endothelium cells were detected by western blot. RESULTS: Administration of naringenin significantly decreased blood pressure, total urine protein level, plasma levels of VCE, α-ADR and angiotensin in PIH mice. Naringenin decreased serum levels of pro-inflammatory cytokines interleukin (IL)-2, IL-6 and tumour necrosis factor alpha (TNF-α), while increased IL-10. Naringenin decreased serum levels of ROS, endothelin while increased SOD and nitric oxide levels. Western blot analysis showed that naringenin inhibited ROS expression, while increased SOD expression in vascular endothelial cells of mice. In addition, western blot also showed that naringenin inhibited JAK2/STAT3 signalling by suppressing SHP-1 expression in vascular endothelial cells of mice. CONCLUSION: Naringenin suppressed the activation of JAK2/STAT3 signalling pathway and promoted SHP-1 expression, leading to ameliorated hypertension in pregnancy.

Efficacy of aspirin combined with labetalol on gestational hypertension and effect on serum PAPP-A, APN and HMGB1.

OBJECTIVE: To determine the clinical efficacy of aspirin combined with labetalol on gestational hypertension (GH) and its influence on serum pregnancy associated plasma protein-A (PAPP-A), adiponectin (APN), and high mobility group box-1 (HMGB1). METHODS: A total of 146 patients with GH admitted to the Zibo Central Hospital between April 2018 and January 2020 were analyzed retrospectively. The control group (Con group, n=71) was treated by labetalol monotherapy, and on this basis, the research group (Res group, n=75) was additionally given aspirin. The following criteria of the 2 groups were evaluated: total effectiveness rate, incidence of adverse reactions, blood pressure-associated indices, coagulation function-associated indices, changes in serum PAPP-A, APN, and HMGB1 levels, adverse maternal and infant outcomes, and neonatal Apgar score. RESULTS: After therapy, the Res group showed a notably higher total effective rate than the Con group, with no remarkable difference in the incidence of adverse reactions. Additionally, compared to the Con group, the Res group showed notably lower systolic blood pressure (SBP) and diastolic blood pressure (DBP), greatly improved coagulation function, significantly lower levels of serum PAPP-A and HMGB1, and significantly higher level of serum APN. Moreover, the incidence of adverse maternal and infant outcomes in the Res group was much lower than that in the Con group after therapy, and the one- and five-min Apgar scores of neonates in the Res group were both notably higher than those in the Con group after delivery. CONCLUSION: For patients with GH, aspirin combined with labetalol can effectively control blood pressure, improve coagulation function and clinical efficacy, lower serum PAPP-A and HMGB1, and increase serum APN, and ameliorate maternal and infant outcome.

The bioflavonoid troxerutin prevents gestational hypertension in mice by inhibiting STAT3 signaling.

Gestational hypertension is a high-risk disease for women, and the current treatments have limited efficacies. Here, we aimed to evaluate troxerutin, which is a natural monomer of flavone, in the treatment of gestational hypertension. Pregnant mice with or without pregnancy-induced hypertension (PIH) were treated with troxerutin (20 and 40 mg/kg) or vehicle. Blood pressure and proteinuria were monitored during treatment. The expression of vasodilation converting enzyme (VCE), angiotensin, TNFα, IL-6, IL-1ß and IL-10 was measured by enzyme-linked immunosorbent assay (ELISA). Oxidative stress was assessed by measuring the reactive oxygen species (ROS) levels and antioxidant enzyme concentrations. Western blot analysis was used to assess the expression of p-STAT3, STAT3, SHP-1, and RNF6. Troxerutin reduced blood pressure and the expression of VCE, angiotensin, urinary protein and pro-inflammatory cytokines in a dose-dependent manner while increasing the expression of anti-inflammatory cytokines. The levels of ROS were decreased, and the levels of antioxidant enzymes were increased. Troxerutin treatment significantly suppressed STAT3/RNF6 signaling. Overexpression of RNF6 attenuated the effects of troxerutin in ameliorating inflammation and oxidative stress. Our data support the use of troxerutin for reducing gestational hypertension due to the role of troxerutin in reducing inflammation and oxidative stress.

Regulative effect of maternal serum fatty acid-binding protein 4 on insulin resistance and the development of gestational diabetes mellitus.

Fatty acid binding protein 4 (FABP4) was found to be closely correlated with gestational diabetes mellitus (GDM), a severe pregnancy syndrome. However, safe and efficient treatment for GDM is limited. We aimed to investigate whether inhibition of FABP4 could ameliorate GDM and the underlying mechanism. An evaluation of blood samples from a total of 109 patients showed significantly positive correlations between serum FABP4 and biochemical parameters known to associate with GDM. This correlation was subsequently explored in vitro. FABP4 inhibition was achieved using BMS309403 in GDM mice. GDM related symptoms, including insulin resistance and macrophage infiltration in the adipose tissues, were measured. Lipid metabolism in 3T3-L1 adipocytes was tested. We firstly confirmed the positive correlations between serum FABP4, insulin resistance and inflammation cytokines, including tumor necrosis factor-α (TNF) and interleukin-6 (IL-6), in GDM patients. Surprisingly, inhibition of FABP4 by BMS309403 resulted in significant alleviation of GDM symptoms in GDM mouse model. BMS309403 improved glucose and insulin tolerance and transcriptionally repressed the levels of TNF-α and IL-6, suggesting a role of FABP4 in inflammation. Furthermore, macrophage infiltration into the adipose tissues was dramatically decreased in the BMS309403-treated GDM mice compared to untreated GDM mice. Interestingly, incubation of 3T3-L1 adipocytes with FABP4 protein decreased the mRNA and protein levels of peroxisome proliferator-activated receptor γ (PPARγ), which was absent when BMS309403 was used. However, lipid accumulation was promoted in FABP4-treated 3T3-L1 adipocytes which showed no change in the presence of BMS309403. In conclusion, inhibition of FABP4 by BMS309403 could be an effective treatment to alleviate GDM.

The isoflavonoid calycosin inhibits inflammation and enhances beta cell function in gestational diabetes mellitus by suppressing RNF38 expression.

BACKGROUND: Gestational diabetes mellitus (GDM) is a medical complication and metabolic disorder associated with pregnancy. Calycosin is a traditional Chinese herbal medicine that is used for the treatment of multiple diseases. This study focused on exploring the effects and underlying mechanisms of Calycosin on GDM. METHODS: The db/+ diabetic mice model of GDM was used to evaluate the effects of calycosin administration on the symptoms of GDM mice. Blood glucose, cytokine production (interleukin 6, IL-6; tumor necrosis factor-α, TNF-α), and insulin levels were measured by ELISA assay. The expression level of signal transducer and activator of transcription 3 (STAT3), ring finger protein 38 (RNF38), and SH2-containing protein tyrosine phosphatase 1 (SHP-1) were determined by Western Blot assay. Beta cell proliferation was assessed by CCK-8 assay. RESULTS: Our data indicated that administration of calycosin significantly improved the GDM symptoms in pregnant db/+ mice as demonstrated by reduced blood glucose, TNF-a, and IL-6 levels as well as increased insulin level, and body weight. Furthermore, we revealed that RNF38/SHP-1/STAT3 signaling should play a critical role in calycosin-promoted beta cell function, and forced expression of RNF38 attenuated the positive effects of calycosin on beta cells. CONCLUSION: Our study implied that calycosin exerts favorable effects on GDM mice via rebalancing insulin sensitivity and inflammatory response.

Serum Levels of TNF-α and IL-6 Are Associated With Pregnancy-Induced Hypertension.

Tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) are proinflammatory cytokines and known to be involved in many pathological processes. However, the association between serum levels of TNF-α, IL-6, and pregnancy-induced hypertension (PIH) is unclear. The aim of the present study was to determine the serum levels of TNF-α and IL-6 and to investigate their potential correlation with PIH. In this study, the serum concentrations of TNF-α and IL-6 in pregnant women who developed PIH and normal pregnant women were measured. We found that the serum concentrations of TNF-α and IL-6 were significantly increased in the patients with PIH compared to the normal pregnant women. In addition, elevated TNF-α and IL-6 concentrations were associated with pathological complications. Moreover, in a hypoxia-induced PIH mice model, animals from the PIH group demonstrated higher TNF-α and IL-6 levels when compared to control, and serum TNF-α and IL-6 levels were positively correlated with right ventricular systolic blood pressure. Furthermore, TNF-α and IL-6 levels were decreased when the PIH mice were treated with remodulin compared to control group. In conclusion, our results suggested that high serum TNF-α and IL-6 levels are associated with PIH, and TNF-α and IL-6 might be potential predictors in the prognosis of PIH.

Serum Beta-Trace Protein as a Novel Predictor of Pregnancy-Induced Hypertension.

Beta-trace protein (BTP) has emerged as a novel biomarker of cardiovascular risk. However, the level of circulating BTP in pregnancy-induced hypertension (PIH) is still unknown. The aim of this study was to determine the concentration of serum BTP in healthy pregnant women and patients with PIH. No significant difference was found in the serum concentration of BTP in patients with a normal pregnancy. In contrast, serum BTP levels in women with PIH (n=46) were significantly higher than those in women with normal pregnancy (n=57). Receiver operating characteristic analysis revealed that using a serum BTP value of 321.3 ng/mL as a cutoff produced a sensitivity of 91.3% and a specificity of 89.5%. Taken together, these findings suggest that a higher serum BTP concentration in PIH patients compared with those with normal pregnancy and serum BTP might be a novel biomarker in the diagnosis of PIH.