Rat Model Of Preeclampsia Research Articles

Metformin inhibits activation of NLRP3 inflammasome and inflammatory response in preeclamptic rats

Backgroud: It is widely acknowledged that Metformin (MET), an established medication for managing type 2 diabetes, possesses diverse pharmacological effects. This study aims to investigate the protective effects of MET against Nω-Nitro-L-arginine methyl ester (L-NAME)-induced preeclampsia (PE). MethodsSprague Dawley (SD) rats were exposed to 200 mg/kg L-NAME with or without prior MET treatment. Histopathological analysis was performed using Hematoxylin and Eosin staining. Serum levels of inflammatory, antiangiogenic, and angiogenic factors were quantified using ELISA kits. Immunohistochemistry (IHC) staining was employed to observe NLRP3 and IL-1β expressions in placental tissues. Western blot and Quantitative Real-Time PCR (q-PCR) analyses were conducted to assess protein and mRNA expressions of NLRP3, caspase-1, ASC, and IL-1β. ResultsWe found that MET could mitigate placental histopathological deterioration and improve pregnancy outcomes in L-NAME-induced PE rat models. MET not only suppressed L-NAME-induced elevation of antiangiogenic factors but also stimulated the production of pro-angiogenic factors. Additionally, MET treatment reversed the excessive inflammatory response induced by L-NAME. Furthermore, MET inhibited the activation of the NLRP3 inflammasome triggered by L-NAME, evidenced by the downregulation of NLRP3 expression, caspase-1, and IL-1β. ConclusionsMET demonstrates a protective effect against L-NAME-induced PE rats, potentially mediated through inhibition of the inflammatory response, downregulation of NLRP3 inflammasome expression in the placenta, and regulation of the balance between anti-angiogenic and pro-angiogenic factors.

Thrombin in Pregnancy and Preeclampsia: Expression, Localization, and Vasoactivity in Brain and Microvessels From Rats.

Thrombin is a coagulation factor increased in pregnancy and further increased in preeclampsia (PE), a hypertensive disorder. Thrombin is also expressed in the brain and may have a nonhemostatic role. We characterized thrombin expression and vasoactivity in brain cerebral parenchymal arterioles (PAs) in rat models of pregnancy and PE. PAs were isolated and pressurized from nonpregnant (NP) and late-pregnant (LP) rats and rats with experimental preeclampsia (ePE). Reactivity to thrombin (1-50 U/mL) was measured in the absence and presence of inhibition of cyclooxygenase and nitric oxide synthase. Plasma levels of prothrombin, thrombin-antithrombin (TAT), tissue plasminogen activator, and plasminogen activator inhibitor-1 (PAI-1) and cerebrospinal fluid levels of TAT were compared using enzyme-linked immunosorbent assay. Expression of protease-activated receptor types 1 and 2 in PAs were measured by Western blot and immunohistochemistry. Neuronal thrombin expression was quantified in brains from all groups by immunohistochemistry. Prothrombin and TAT were elevated in ePE plasma compared with NP and LP. TAT was detected in cerebrospinal fluid from all groups and significantly elevated in LP (NP: 0.137 ± 0.014 ng/mL, LP: 0.241 ± 0.015 ng/mL, ePE: 0.192 ± 0.028 ng/mL; P < 0.05). Thrombin caused modest vasoconstriction in PAs from all groups regardless of cyclooxygenase or nitric oxide synthase inhibition. PAR1 and PAR2 were found in PAs from all groups colocalized to smooth muscle. Thrombin expression in central neurons was decreased in both LP and ePE groups compared with NP. These findings suggest a role for thrombin and other hemostatic changes during pregnancy and PE beyond coagulation.

Ghrelin alleviates placental dysfunction by down-regulating NF-κB phosphorylation in LPS-induced rat model of preeclampsia

In our previous study, we uncovered that ghrelin promotes angiogenesis in human umbilical vein endothelial cells (HUVECs) in vitro by activating the Jagged1/Notch2/VEGF pathway in preeclampsia (PE). However, the regulatory effects of ghrelin on placental dysfunction in PE are unclear. Therefore, we applied Normal pregnant Sprague-Dawley (SD) rats, treated with lipopolysaccharide (LPS), to establish a PE-like rat model. The hematoxylin-eosin (HE) staining method and immunohistochemistry (IHC) technology were used to detect morphological features of the placenta. IHC and Western blot were applied to examine Bax and Bcl-2 expression levels. The concentrations of serum soluble fms-like tyrosine kinase-1 (sFlt1) and placental growth factor (PIGF) were assessed by enzyme-linked immunosorbent assay (ELISA) kit. In addition, the apoptosis rates of JEG-3 and HTR-8/SVneo trophoblast cells were determined by Annexin V-FITC/PI apoptosis detection kit. Cell migratory capacities were assessed by scratch-wound assay, and RNA-sequencing assay was used to determine the mechanism of ghrelin in regulating trophoblast apoptosis. It has been found that ghrelin significantly reduced blood pressure, urinary protein, and urine creatinine in rats with PE, at the meanwhile, ameliorated placental and fetal injuries. Second, ghrelin clearly inhibited placental Bax expression and circulating sFlt-1 as well as elevated placental Bcl-2 expression and circulating PIGF, restored apoptosis and invasion deficiency of trophoblast cells caused by LPS in vitro. Finally, transcriptomics indicated that nuclear factor kappa B (NF-κB) was the potential downstream pathway of ghrelin. Our findings illustrated that ghrelin supplementation significantly improved LPS-induced PE-like symptoms and adverse pregnancy outcomes in rats by alleviating placental apoptosis and promoting trophoblast migration.

Protective Effects of Aspirin Supplemented With Quercetin in L-NAME-Induced Preeclampsia-Like Rats.

Aspirin supplemented with quercetin was reported to enhance the therapeutic effects of aspirin in a rat model of preeclampsia. In this study, the underlying mechanisms were further explored. Preeclampsia was induced by L-NAME (50 mg/kg/day) via oral gavage from gestation day (GD)14 to GD19. Aspirin (1.5 mg/kg/day) administration was performed using aspirin mixed with rodent dough from GD0 to GD19. The administration of quercetin (2 mg/kg/day) was performed by intraperitoneal infusion from GD0 to GD19. Protein levels were evaluated using ELISA or Western blot, and microRNA (miRNA) level was evaluated by RT-PCR. Aspirin supplemented with quercetin ameliorated the increase of systolic blood pressure (SBP), proteinuria, tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels, and improved the pregnancy outcomes in preeclampsia rats. Aspirin supplemented with quercetin inhibited miR-155 expression in preeclampsia rats. The decreased miR-155 level in placenta further increased the protein level of SOCS1 and inhibited the phosphorylation of p65. In this study, we demonstrated that aspirin supplemented with quercetin enhanced the effects of aspirin for the treatment of preeclampsia.

Improvement in Clinical Features of L-NAME-Induced Preeclampsia-like Rats through Reduced SERPINA5 Expression.

Pre-eclampsia (PE) is a severe pregnancy disorder that poses a significant health risk to both mother and fetus, with no preventive or therapeutic measures. Our previous research suggested an association between elevated SERPINA5 levels and PE features. This study investigated whether SERPINA5 could be a potential therapeutic target for PE. We established PE-like features in pregnant rats using L-NAME (75 mg/kg/d) treatment. Adenoviruses carrying overexpressed or suppressed SERPINA5 genes were intravenously injected into these PE rats on the fifth and seventh days of pregnancy. We evaluated the rats' systolic blood pressure, urine protein concentration, and placental and fetal metrics and histology. Placental gene expression following SERPINA5 overexpression was evaluated using mRNA sequencing. The L-NAME-induced PE rat model observed a significant increase in placental and peripheral SERPINA5 levels. The overexpression of SERPINA5 exacerbated L-NAME-induced hypertension and proteinuria in pregnant rats. A histology examination revealed a smaller placental junctional zone in L-NAME + overexpressing rats. Placental gene expression analysis in the L-NAME + overexpressing group indicated increased coagulation activation. L-NAME-induced hypertension and proteinuria were mitigated when SERPINA5 expression was suppressed. Additionally, placental development was improved in the SERPINA5-suppressed group. Our findings suggested that SERPINA5 may worsen L-NAME-induced PE-like features by promoting the activation of the coagulation cascade. Therefore, reducing SERPINA5 expression could potentially serve as a therapeutic strategy for PE.

Suppression of circular RNA serum and glucocorticoid-induced kinase 1 elevates antioxidant molecules and angiogenesis in trophoblast cells to attenuate preeclampsia via microRNA-508-3p to target and restrain PUM homolog 1.

Preeclampsia (PE) is a pregnancy-specific syndrome characterized by hypertension and proteinuria. Recently, multiple circular RNAs (circRNAs) were considered latent clinical diagnostic markers or therapeutic targets. This study was to explore the impact of circRNA serum and glucocorticoid-induced kinase 1 (SGK1) on PE via influencing the microRNA (miR)-508-3p/PUM homolog 1 (PUM1) axis. Placental tissues of 34 pregnant women with PE and 34 normal pregnant women were collected to measure circRNA SGK1 (circSGK1), miR-508-3p, and PUM1. Human placental trophoblasts HTR-8/SVneo were transfected with plasmids, thereafter to observe proliferation, migration, invasion, and apoptosis, analyze antioxidant molecules Troxerutin (TXN), Glutamate-cysteine ligase catalytic subunit (GCLC), NAD (P) H-quinone oxidoreductase 1 (NQO1), and determine angiogenesis. After the construction of the PE rat model, antioxidant molecules TXN, GCLC, and NQO1, vascular-associated factor vascular endothelial growth factor A (VEGF-A), and histopathological conditions were tested. Examination of the binding of circSGK1 and miR-508-3p with PUM1 was performed. Our data showed that circSGK1 expression was elevated in the placenta of patients with PE. Silenced circSGK1 or elevated miR-508-3p promoted the growth and antioxidant molecules and angiogenesis in trophoblast cells; CircSGK1 combined with miR-508-3p, and miR-508-3p targeted PUM1. In summary, suppression of circSGK1 augments antioxidant molecules and angiogenesis in trophoblast cells to attenuate PE via miR-508-3p to target PUM1.

The effect of tempol, a potent synthetic antioxidant, on the limb teratogenicity in an experimental model of preeclampsia in rats

The effect of tempol, a potent synthetic antioxidant, on the limb teratogenicity in an experimental model of preeclampsia in rats

Abstract P208: Cardiovascular Effects Of Antihypertensive Agents In A Rat Model Of Preeclampsia.

Introduction: Hypertensive disorders of pregnancy are a leading cause of maternal and neonatal morbidity and mortality. The placental origin of preeclampsia, characterized by hypertension and proteinuria after 20 weeks of gestation, is currently on the test bench. A number of studies reported a cardiovascular origin for preeclampsia, with increased long-term cardiovascular risk for affected women. Despite new insights into pathophysiology, there is a lack of evidence supporting improvement in maternal and neonatal outcomes with current antihypertensive therapy. This study aims to investigate maternal hemodynamics and cardiovascular parameters in a preeclamptic rat model, focusing on changes induced by antihypertensive treatment. Purpose: This study challenges blood pressure as the sole target for antihypertensive therapy in preeclampsia, seeking new insights into the disease's pathophysiology through invasive procedures and assessment of cardiac morphology in a rat model for preeclampsia. Methods: Maternal hemodynamics were invasively measured, and cardiac compliance was assessed via echocardiography. Cardiac phenotyping was performed combining the evaluation of physiology and structural changes. Maternal hearts were investigated at the end of pregnancy and postpartum. Blood pressure was continuously monitored using telemetry. The antihypertensive agents used in the study were methyldopa, nifedipine, and labetalol, which represent standard therapy for women with preeclampsia. Results: In the transgenic rat model for preeclampsia (PE), treatment with methyldopa and nifedipine led to a reduction of blood pressure during gestation whereas labetalol had no effect on the blood pressure (mean arterial pressure: PE 160.80±3.85 mmHg; PE+methyldopa 145.01±3.71 mmHg; PE+nifedipine 122.08±5.62 mmHg; PE+labetalol 163.12±3.12 mmHg). Despite the antihypertensive effects of methyldopa and nifedipine, the maternal hemodynamics as well as the cardiac compliance were unimproved during and after pregnancy. Conclusion: The results showed that standard therapy for women with preeclampsia does not seem to be sufficient to prevent the cardiovascular changes and therefore to reduce the CDV risk.

PO3_05. Maternal b cell depletion reduces blood pressure and improves birth weights in a rat model of preeclampsia

PO3_05. Maternal b cell depletion reduces blood pressure and improves birth weights in a rat model of preeclampsia

Abstract P200: Tnf-α Blockade During Pregnancy Does Not Impact Offspring Development In A Rat Model Of Preeclampsia

Preeclampsia (PE), new onset hypertension during pregnancy, is associated with increased tumor necrosis factor alpha (TNF-α), agonistic autoantibodies to the angiotensin II type 1 receptor (AT1-AA), and low birth weight babies. We have previously shown that Etanercept ( E ), a TNF-α inhibitor, improves PE features in the reduced uterine perfusion pressure (RUPP) rat. However, the long-term effect of E on offspring health and immune competency has not been studied. Therefore, we hypothesize that E will improve maternal health without affecting offspring outcomes in a rat model of PE. To test this hypothesis, E (0.4 mg/kg) was injected on gestation day 18 into sham and RUPP dams which were allowed to deliver. Weights were recorded at birth and 16 weeks of age when mean arterial pressure (MAP) was measured. Whole blood was collected to measure immune cells by flow cytometry. AT1-AA was measured using a cardiomyocyte bioassay. A student’s t-test was used to compare differences among male and female RUPP offspring with and without E . RUPP male and female offspring were smaller (5.21±0.19 g, n=7; 5.24±0.10 g, n=7) than sham offspring (5.94±0.14 g, n=7, p&lt;0.05; 5.86±0.16 g, n=9, p&lt;0.01) while RUPP male offspring were larger and female offspring were unchanged following perinatal E (6.01±0.29 g, n=9, p&lt;0.05; 5.28±0.14, n=9). MAP was comparable between sham (133±4 mmHg) and RUPP (132±4 mmHg) male offspring with no change seen following perinatal E (133±4 mmHg). Perinatal E did not impact adult male circulating B cells. Circulating AT1-AA was elevated in male RUPP offspring (16±2 ΔBPM, p&lt;0.01) compared to sham and was not improved with E (13±2 ΔBPM). MAP was comparable in sham (131±5 mmHg) and RUPP (130±5 mmHg) female offspring but trended downward with perinatal E (123±4 mmHg). RUPP female offspring had reduced circulating B cells (4.24±0.81 % gated, p=0.066) compared to sham females (9.56±2.07 % gated) which was normalized following perinatal E (9.04±1.42 % gated, p&lt;0.05). AT1-AA trended lower following perinatal E (7±2 ΔBPM, p=0.0888) compared to RUPP female offspring (16±3 ΔBPM). Our findings indicate that E does not worsen fetal outcomes. Moreover, E improves male birth weight and reduces adult female AT1-AA without compromising offspring immune competency.

Abstract P420: Changes In Cardiac Oxidative Stress Is Protected In Intrauterine Growth Restricted Females Vs Males Before The Development Of Hypertension And Cardiovascular Diseases

Preeclampsia (PE) is a pregnancy complication characterized by hypertension (HTN) typically during the third trimester. PE is the leading cause for intrauterine growth restricted (IUGR) offspring (OFS). Women with PE and their OFS (IUGR) are at a higher risk for developing cardiovascular diseases (CVD) later in life. Two major factors that contribute to the development of CVDs are HTN and oxidative stress (OS). Studies show that human and animals adult IUGR OFS have a higher risk of developing HTN and CVDs compared to females later in life. The rationale for these sex differences is unknown; and we hypothesize that male IUGR rats will have elevated cardiac oxidative stress, while female IUGR will have lower cardiac OS .To test, we will examine sex differences between 12 week IUGR and control (CON) OFS that are not hypertensive, to determine if there are any change in cardiac OS before the development of HTN and CVD. Methods: Sprague Dawley rats were divided into 4 groups: IUGR male, IUGR female, and control (CON) male and female. IUGR rats were derived from the OFS reduced uterine perfusion pressure (RUPP) PE rat model. CON OFS were from normal pregnant rats. At 12wk, BP was measured via carotid catheterization. Heart tissue was collected to assess oxidative stress, via examining reactive oxygen species (ROS) Heat shock protein 1 (HSP-1) and hydrogen peroxide (H2O2). Antioxidants were measured via manganese superoxide dismutase (MnSOD), copper zinc superoxide dismutase (CuZnSOD) and antioxidant capacity (AC). There were no significant changes in HSP-1, CuZnSOD, and H2O2 between male and female OFS. However, female IUGR OFS had a 20% increase in antioxidant MnSOD (119.9 ± 6.133 vs 93.98 ± 5.197 %MnSOD U/protein/CON, p &lt;0.05) compared to CON in the heart. Male OFS had no significant changes for MnSOD. IUGR female AC had a 2-fold increase in activity (22.13 ± 1.019 vs 14.15 ± 1.852 mM Trolox/mg protein, p &lt;0.05) compared to its CON. Furthermore, IUGR male OFS had a slight decrease in AC compared to CON male (11.00 ± 1.420 vs 13.24 ± 0.4676 mM Trolox/mg protein, ns). The significant increase of AC and MnSOD in IUGR female OFS suggests that females may be more protected from OS, via mitochondria-specific antioxidants compared to males.

Abstract 062: Lower Middle Cerebral Artery Resistance In A Transgenic Rat Model Of Preeclampsia: A Platform For Studying Long-term Preeclampsia Induced Cerebrovascular Disease

Preeclampsia (PE) is a pregnancy disorder characterized by new onset hypertension and end-organ dysfunction. PE patients have increased risk of cerebrovascular disease (CVD) later in life; however, the mechanisms are unclear. We determined whether exposure to a PE-like syndrome in transgenic rat (TGA-PE: female TG for angiotensinogen mated to male TG for renin) alters left middle cerebral artery (LMCA) or left internal carotid (LICA) artery hemodynamics at 2 or 5 months post-pregnancy. The analyses of systolic (SBP) and diastolic (DBP) blood pressures and heart rate (HR) (by tail cuff; NIBP-8 Columbus Instruments), and LMCA and LICA pulsatility index (PI) (by transcranial Doppler (TCD); Vevo 2100), were performed at late gestation, and at 2 and 5 months post-pregnancy. SBP and DBP were elevated in TGA-PE vs. Sprague-Dawley (SD) in late pregnancy (n=5-6), and at 2 (n=4-6), and 5 months (n=2-3) post-pregnancy (p&lt;0.05). HR was elevated in TGA-PE vs. SD in late pregnancy (493.5±10.6 vs. 392.5±14.2, p&lt;0.05, n=5-6), but not at 2 or 5 months post-pregnancy. However, LMCA PI was lower in TGA-PE vs. SD at 2 months post-pregnancy (p&lt;0.05), and tended to decrease in late gestation and at 5 months post-pregnancy. LICA PI was lower in TGA-PE vs. SD during pregnancy (0.729 ± 0.086 vs 1.34 ± 0.095, p&lt;0.05, n=4-5). In summary, PE-like syndrome in TGA-PE resulted in higher SBP and lower LMCA resistance versus SD at 2 months post-pregnancy, suggesting a dysfunctional autoregulation of cerebral blood flow. Since women with history of PE have lower MCA resistance, we conclude that TGA-PE model may provide a suitable platform to investigate the mechanistic link between PE and future development of CVD.

Abstract P203: Microvascular Imaging As Early Predictor For Cardiac Dysfunction After Preeclampsia

Preeclampsia (PE), the most common pregnancy disease, is the main cause for maternal and fetal mortality and morbidity worldwide. The disease is characterized by a sudden increase in blood pressure in combination with signs of end organ damage, e.g. proteinuria and the underlying mechanisms are poorly understood. Immune dysregulation and endothelial dysfunction are important pathomechanisms leading to PE, we hypothesize that anti-angiogenesis and dysregulation of microvascular perfusion postpartum lead to the increased cardiovascular risk in later life. In addition, we will test if the examination of retinal vessels can be used as a diagnostic window for early risk prediction of cardiovascular alterations. In a well-established transgenic rat model for PE, we performed speckle tracking echocardiography, immunostaining, RNA sequencing and small vessel imaging by micro-computed tomography of the heart and the eyes in former preeclamptic rats (PEpp) and healthy controls (WTpp). A reduction of cardiac vessels (AUC 0-200 μm: WTpp 77.19±4.64 vs. PEpp 62.59±2.72) in combination with an increase of small vessels in the retina (AUC 0-50 μm: WTpp 52.89±1.47 vs. PEpp 59.54±1.09) is shown postpartum. Data from echocardiography (Global longitudinal strain: WTpp -22.03±2.47 % vs. PEpp -14.50±1.80 %) and histological staining (CD31: WTpp 292.7±30.9 per fov vs. PEpp 219.0±10.5 per fov) support the hypothesis of persistent cardiovascular alterations after PE. Performing RNAseq, 65 genes were found to be permanently altered in the left ventricle of the heart of preeclamptic rats compared to 4 genes in controls. In the retinal tissue, 39 genes in PEpp and no genes in WTpp were identified postpartum compared to never pregnant animals (adjusted p-value &lt; 0.05,

Hesperidin improves physiological outcomes in an arginine vasopressin rat model of pre-eclampsia.

Hesperidin, a flavanone commonly found in citrus fruits and herbal formulations, has emerged as a potential new therapeutic agent for modulating several diseases. Since pre-eclampsia is a growing public health threat, it may negatively impact the economy and increase the disease burden of South Africa. Phytocompounds are easily accessible, demonstrate minimal side effects, and may confer novel medicinal options as a treatment and preventive preference. To investigate the physiological, biochemical, and hematological outcomes of hesperidin in an arginine vasopressin (AVP)-induced rodent model of pre-eclampsia. Female Sprague-Dawley rats were surgically implanted with mini-osmotic pumps to deliver AVP (200 ng/h) subcutaneously. Animals were treated with hesperidin at 200 mg/kg.b.w via oral gavage for 14 days. Systolic and diastolic blood pressures were measured on GD 7, 14, and 18 using a non-invasive tail-cuff method and were euthanized on GD 21. The findings showed that hesperidin administration significantly decreased blood pressure (P < 0.05) and urinary protein levels in pregnant rats (P < 0.001). Placental and individual pup weight also increased significantly in the pregnant hesperidin-treated groups compared to AVP untreated groups (P < 0.001). Biochemical and hematological markers such as white blood cell count and lymphocyte levels differed significantly (P < 0.05) in AVP groups treated with and without hesperidin. Our results suggest that hesperidin is an antihypertensive agent with modes of action associated with its diuretic and blood pressure lowering effects and reduction of proteinuria in AVP-induced pre-eclamptic rats.

HOXB3 promotes trophoblast cell proliferation, invasion, and migration to alleviate preeclampsia via mediating the Notch/Wnt/β-catenin pathway

Preeclampsia (PE) harms a significant number of pregnant women and fetuses. However, because of its complex pathological mechanisms, there is no cure except for delivery. This study identified the impact and mechanisms of action of HOXB3 in PE. The behaviors of HTR-8/SVneo cells were analyzed using a cell counting kit-8, EdU, and transwell assays. The interaction between HOXB3 and Notch1 was assessed using a luciferase reporter and chromatin immunoprecipitation assays. Expression was measured by quantitative real-time polymerase chain reaction, western blotting, and immunofluorescence assays. Additionally, the function of HOXB3 was evaluated in an established rat model of PE. We found that HOXB3 was upregulated in PE. HOXB3 overexpression facilitated trophoblast cell proliferation, migration, and invasion. HOXB3 transcriptionally regulated Notch1 by binding to its promoter. Notch1 knockdown abrogated the functions of HOXB3 and the-catenin pathway in trophoblasts. Suppression of the Wnt/β-catenin pathway abrogated the effects of HOXB3. Additionally, HOXB3 alleviated the symptoms in PE rats. In conclusion, HOXB3 transcriptionally activated Notch1 expression and the-catenin pathway, promoting trophoblast cell proliferation, invasion, and migration, thereby alleviating PE progression. This study provides a novel approach for PE therapy.

Altered offspring neurodevelopment in an L-NAME-induced preeclampsia rat model.

To investigate the mechanism underlying the increased risk of subsequent neurodevelopmental disorders in children born to mothers with preeclampsia, we evaluated the neurodevelopment of offspring of a preeclampsia rat model induced by the administration of N-nitro-L-arginine methyl ester (L-NAME) and identified unique protein signatures in the offspring cerebrospinal fluid. Pregnant rats received an intraperitoneal injection of L-NAME (250 mg/kg/day) during gestational days 15-20 to establish a preeclampsia model. Behavioral experiments (negative geotaxis, open-field, rotarod treadmill, and active avoidance tests), immunohistochemistry [anti-neuronal nuclei (NeuN) staining in the hippocampal dentate gyrus and cerebral cortex on postnatal day 70], and proteome analysis of the cerebrospinal fluid on postnatal day 5 were performed on male offspring. Offspring of the preeclampsia dam exhibited increased growth restriction at birth (52.5%), but showed postnatal catch-up growth on postnatal day 14. Several behavioral abnormalities including motor development and vestibular function (negative geotaxis test: p < 0.01) in the neonatal period; motor coordination and learning skills (rotarod treadmill test: p = 0.01); and memory skills (active avoidance test: p < 0.01) in the juvenile period were observed. NeuN-positive cells in preeclampsia rats were significantly reduced in both the hippocampal dentate gyrus and cerebral cortex (p < 0.01, p < 0.01, respectively). Among the 1270 proteins in the cerebrospinal fluid identified using liquid chromatography-tandem mass spectrometry, 32 were differentially expressed. Principal component analysis showed that most cerebrospinal fluid samples achieved clear separation between preeclampsia and control rats. Pathway analysis revealed that differentially expressed proteins were associated with endoplasmic reticulum translocation, Rab proteins, and ribosomal proteins, which are involved in various nervous system disorders including autism spectrum disorders, schizophrenia, and Alzheimer's disease. The offspring of the L-NAME-induced preeclampsia model rats exhibited key features of neurodevelopmental abnormalities on behavioral and pathological examinations similar to humans. We found altered cerebrospinal fluid protein profiling in this preeclampsia rat, and the unique protein signatures related to endoplasmic reticulum translocation, Rab proteins, and ribosomal proteins may be associated with subsequent adverse neurodevelopment in the offspring.

H3K27me3-modulated Hofbauer cell BMP2 signalling enhancement compensates for shallow trophoblast invasion in preeclampsia.

Preeclampsia (PE) is a common hypertensive pregnancy disorder associated with shallow trophoblast invasion. Although bone morphogenetic protein 2 (BMP2) has been shown to promote trophoblast invasion invitro, its cellular origin and molecular regulation in placenta, as well as its potential role in PE, has yet to be established. Additionally, whether BMP2 and/or its downstream molecules could serve as potential diagnostic or therapeutic targets for PE has not been explored. Placentas and sera from PE and healthy pregnant women were subjected to multi-omics analyses, immunoblots, qPCR, and ELISA assays. Immortalized trophoblast cells, primary cultures of human trophoblasts, and first-trimester villous explants were used for invitro experiments. Adenovirus expressing sFlt-1 (Ad Flt1)-induced PE rat model was used for invivo studies. We find globally decreased H3K27me3 modifications and increased BMP2 signalling in preeclamptic placentas, which is negatively correlated with clinical manifestations. BMP2 is derived from Hofbauer cells and epigenetically regulated by H3K27me3 modification. BMP2 promotes trophoblast invasion and vascular mimicry by upregulating BMP6 via BMPR1A-SMAD2/3-SMAD4 signalling. BMP2 supplementation alleviates high blood pressure and fetal growth restriction phenotypes in Ad Flt1-induced rat PE model. Our findings demonstrate that epigenetically regulated Hofbauer cell-derived BMP2 signalling enhancement in late gestation could serve as a compensatory response for shallow trophoblast invasion in PE, suggesting opportunities for diagnostic marker and therapeutic target applications in PE clinical management. National Key Research and Development Program of China (2022YFC2702400), National Natural Science Foundation of China (82101784, 82171648, 31988101), and Natural Science Foundation of Shandong Province (ZR2020QH051, ZR2020MH039).

Use of Traditional and Proteomic Methods in the Assessment of a Preclinical Model of Preeclampsia.

Recent studies have demonstrated downregulation of breast cancer resistance protein (BCRP/ABCG2) in placenta obtained from women with preeclampsia (PE). BCRP is highly expressed in placenta and plays an important role in preventing xenobiotics from entering the fetal compartment. While PE is often therapeutically managed with drugs that are substrates of BCRP, there are limited studies on the impact of PE on fetal drug exposure. Due to ethical concerns, use of preclinical models is an important approach. Thus, by using proteomic and traditional methods, we characterized transporter changes in an immunological rat model of PE to determine its utility and predictive value for future drug disposition studies. PE was induced by daily administration of low-dose endotoxin (0.01-0.04 mg/kg) to rats on gestational days (GD) 13 to 16, urine was collected, and rats were sacrificed on GD17 or GD18. PE rats shared similar phenotype to PE patients, including proteinuria, and increased levels of TNF-α and IL-6. Transcript and protein levels of Bcrp were significantly downregulated in placenta of PE rats on GD18. Mdr1a, Mdr1b and Oatp2b1 mRNA were also decreased in PE. Proteomics revealed activation of various hallmarks of PE including immune activation, oxidative stress, endoplasmic reticulum stress and apoptosis. Overall, our results demonstrated that the immunological PE rat model exhibits numerous similarities to human PE along with dysregulation of placental transporters. Therefore, this model may be useful in examining the impact of PE on the maternal and fetal disposition of BCRP substrates. Significance Statement Fully characterizing preclinical models of disease is necessary in order to determine their validity to human conditions. Combining traditional and proteomic methods of model characterization, we identified numerous phenotypic similarities between our model of PE and human disease. The alignment with human pathophysiological changes allows for more confident use of this preclinical model.

Natural Flavonoid Quercetin Enhances the Anti-inflammatory Effects of Aspirin in a Preeclampsia-like Rat Model Induced by Lipopolysaccharide.

Preeclampsia is a disorder of hypertension and proteinuria accompanied by abnormal inflammatory responses. Both aspirin and quercetin possess anti-inflammatory and anti-hypertensive properties. A low dose of aspirin is recommended for the prevention of preeclampsia in patients with preeclampsia history. Whether quercetin can enhance the effect of aspirin on preeclampsia remains elusive. Female Sprague-Dawley pregnant rats were treated with daily administration of aspirin, quercetin, or a combination of aspirin and quercetin and subsequently received lipopolysaccharides (LPS) injection to induce preeclampsia-like symptoms. The systolic blood pressure and proteinuria from all groups of rats were assessed. Our results demonstrated that the combination of quercetin and aspirin exerted significantly stronger effects than aspirin alone on decreasing systolic blood pressure and proteinuria, reducing pro-inflammatory cytokine production, and inhibiting M1-type decidual macrophages polarization in an LPS-induced rat model of preeclampsia. This study suggested that quercetin may serve as an excellent supplement to aspirin in preventing or treating patients with preeclampsia.

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.