Background White matter injury (WMI) induced by intrauterine inflammation can cause adverse neurological outcomes. Fibrinogen-like protein 2 (FGL2)/fibroleukin is an important trigger of inflammatory responses and is involved in some cerebral diseases. However, the role of FGL2 in intrauterine inflammation-induced WMI remains unclear. Methods Lipopolysaccharide (LPS) was intraperitoneally injected into wild-type and FGL2 knockout mice to induce intrauterine inflammation. Body weight and brain weight of offspring were monitored. Major basic protein (MBP) expression was evaluated to demonstrate the myelination of offspring. To investigate the regulatory mechanism of FGL2, cytokine expression, microglial polarization, and the activation of mitogen-activated protein kinase (MAPK) signaling pathway in the offspring were analyzed. Results Upon LPS exposure, FGL2 knockout offspring showed a significant increase in body weight loss. MBP reduction induced by LPS was prevented in FGL2 knockout offspring. Expression levels of proinflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α, and M1 marker CD86 were suppressed, while the expression levels of anti-inflammatory cytokines IL-10 and M2 marker CD206 were increased. FGL2 deficiency significantly inhibited the phosphorylation of p38MAPK and c-Jun N-terminal kinase (JNK) protein. Conclusions FGL2 deficiency can ameliorate WMI induced by intrauterine inflammation, reducing inflammatory cascade and improving hypomyelination, through the regulation of microglial polarization and MAPK signaling pathways. Impact Intrauterine inflammation induces WMI leading to severe neurological sequelae. FGL2 plays an important role in the progression of WMI induced by intrauterine inflammation.FGL2 deficiency can protect against WMI by inhibiting p38 MAPK and JNK phosphorylation, regulating microglia polarization, and reducing inflammation response.FGL2 could be a novel molecular target for protecting against WMI induced by intrauterine inflammation.

Background: Congenital nephrogenic diabetes insipidus (CNDI) is a rare inherited disease that is caused by mutations in arginine vasopressin receptor 2 (AVPR2) or aquaporin 2 (AQP2). Functional analysis of the mutated receptor is necessary to verify the impact of the mutation on receptor function and suggest some possible therapeutic strategies for specific functional defects.Methods: Family history and clinical information were collected. Whole-exome sequencing and sanger sequencing were performed to determine the potential genetic cause of diabetes insipidus. The identified variant was classified according to the American College of Medical Genetics (ACMG) criteria. Bioinformatic analysis was performed to predict the function of the identified variation. Moreover, wild-type and mutated AVPR2 vectors were constructed and transfection to HEK-293T cells. Immunofluorescence experiments were performed to investigate the expression and localization of the mutated protein and cAMP parameter assays were used to measure its activity in response to AVP.Results: The heights of the adult members affected with polyuria and polydipsia were normal, but all affected children had growth retardation. Next-generation sequencing identified a novel mutation in AVPR2 gene (c.530T > A) in this family. Bioinformatic analysis indicated that the mutation in AVPR2 changed the hydropathic characteristic of the protein and was probably deleterious. Although immunofluorescence showed that the mutated AVPR2 was normally expressed in the cell surface, the intracellular cAMP concentration stimulated by AVP was significantly lower in cells transfected with mutated AVPR2 than cells transfected with wild-type AVPR2. Based on the ACMG criteria, the novel c.530T > A variant of the AVPR2 gene was likely pathogenic and the affected family members were diagnosed as CNDI. After the confirmation of the diagnosis, the proband was treated with compound amiloride hydrochloride and rhGH, the symptoms of polyuria, polydipsia and growth retardation were all improved.Conclusion: These findings suggested that the novel mutation in AVPR2 (c.530T > A) was a true disease-causing variant with mild effects, which could be classified as a type III mutant receptor. Moreover, investigations of the function of growth hormone axis could be important for the pediatric CNDI patients with extreme short stature, and rhGH treatment might improve the final adult heights in these patients.

Background: To explore whether maternal obesity inhibits placental angiogenesis through downregulation of Sirtuin 1/Peroxisome proliferator-activated receptor-γ coactivator-1α (SIRT1/PGC-1α) signaling pathway.Methods: In a rat model of pre-pregnancy obesity, rats were sacrificed at embryonic day (E)18.5. Maternal characteristics were measured. Placentas were collected to observe the pathological changes and angiogenesis using hematoxylin-eosin (HE) staining and platelet endothelial cell adhesion molecule-1 [PECAM-1/CD31 (CD31)] immunohistochemical (IHC) staining, and the expression of the SIRT1/PGC-1α signaling pathway was also analyzed using western blotting and quantitative real-time polymerase chain reaction (qPCR). In in vitro experiments, human umbilical vein endothelial cells (HUVECs) were incubated under high fat conditions. We activated and inhibited the SIRT1/PGC-1α signaling pathway to determine the proliferation, angiogenic tube formation, and migration capacity of endothelial cells. Cell counting kit-8 (CCK-8) assays, tubule formation assays, and scratch wound-healing migration assays were also performed. Results:In vivo results showed that compared with the control group, the high-fat diet (HFD) group were heavier and their plasma triglyceride and total cholesterol contents were higher. The ratio of fetal weight to placental weight was reduced in the HFD group compared to the control group. In the HFD group, placental angiogenesis was decreased, and the SIRT1/PGC-1α signaling pathway was down-regulated compared with that in the control group. The results of in vitro experiments showed that SRT1720 reduced SIRT1/PGC-1α and vascular endothelial growth factor (VEGFA) expression inhibition induced by high fat stress, while EX-527 increased SIRT1/PGC-1α and VEGFA expression inhibition. Compared with the control group, maternal obesity impaired placental angiogenesis and reduced the proliferation and migration of HUVECs. Conclusions:The results suggest that maternal obesity impairs placental angiogenesis. They also provide experimental evidence that activation of the SIRT1/PGC-1α signaling pathway improves angiogenesis in vitro.