YKL-40 promotes proliferation and invasion of HTR-8/SVneo cells by activating akt/MMP9 signalling in placenta accreta spectrum disorders.

YKL-40 is a secreted glycoprotein that can promote invasion, angiogenesis and inhibit apoptosis, and was highly expressed in a variety of tumours. In this paper, we investigated the impacts of YKL-40 on proliferation and invasion in HTR-8/SVneo cells during placenta accreta spectrum disorders (PAS) development. The levels of YKL-40 protein in late-pregnant placental tissue were detected using immunohistochemistry and Western blotting, and gene expression using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The proliferation, migration, invasion and apoptosis abilities of HTR-8/SVneo cells were detected by cell counting kit-8 (CCK-8), Transwell, scratch assay, and flow cytometry, respectively. Our current results showed that YKL-40 was significantly increased in the PAS group compared to the normal control group (P < 0.01). Biological function experiments showed that YKL-40 significantly promoted the proliferation, migration and invasion of HTR-8/SVneo cells, and inhibited cell apoptosis. Knockdown of YKL-40 inhibited the activation of Akt/MMP9 signalling in trophoblast cells. These data suggested that YKL-40 might be involved in the progression of PAS, which may be attributed to the regulation of Akt/MMP9 signalling pathway.

What is already known on this subject? YKL-40 is a secretory glycoprotein that can promote invasion, angiogenesis, and inhibit apoptosis and was highly expressed in a variety of tumours. Trophoblast cells resemble tumour cells in their migration and invasion.What the results of this study add? YKL-40 expression was significantly up-regulated in PAS. CCK-8 assays showed that YKL-40 remarkably enhanced the viability of HTR-8/SVneo cells. Scratch and Transwell assays demonstrated that YKL-40 significantly promoted the migration and invasion of HTR-8/SVneo cells. Additionally, YKL-40 attenuated apoptosis in HTR-8/SVneo cells.What the implications are of these findings for clinical practice and/or further research? Akt/MMP9 was involved in the regulation of YKL-40 on trophoblast invasion, which may provide theoretical basis and new ideas for the drug blocking intervention of placenta accreta.

Effects of blood glucose on vaspin secretion in patients with gestational diabetes mellitus.

OBJECTIVE: This study aimed to observe the changes in serum vaspin levels in pregnant women after glucose load during oral glucose tolerance test (OGTT) and analyze the effect of blood glucose levels on serum vaspin secretion in pregnant women with gestational diabetes mellitus (GDM). METHODS: It included 30 patients with GDM and 30 age-matched pregnant women with normal glucose tolerance (NGT, control). The blood glucose, insulin, total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL), and serum vaspin levels were measured at 24-28 weeks of gestation. The homeostasis model assessment of insulin resistance (HOMA-IR) values were calculated. RESULTS: 1. The levels of fasting plasma glucose (FPG), 1-h PG, 2-h PG, fasting insulin (FINS), HOMA-IR, baseline vaspin, 1-h vaspin, and 2-h vaspin were higher in the GDM group than in the NGT group. 2. The 1-h and 2-h vaspin levels were significantly higher than the baseline levels in the GDM group. The change in the vaspin level after 1 h in the GDM group during the OGTT was significantly greater than that in the NGT group. 3. The vaspin levels in the GDM group positively correlated with 1-h PG and TG, including the baseline vaspin (r = 0.494, p = .006; r = 0.385, p = .036), 1-h vaspin (r = 0.577, p = .001; r = 0.466, p = .010), and 2-h vaspin (r = 0.557, p = .001; r = 0.488, p = .006). CONCLUSION: Vaspin might be regulated by the hyperglycemia level and involved in the pathogenesis of GDM and lipid metabolism.

[Role of visfatin in the pathogenesis of gestational diabetes mellitus and its relationship with insulin resistance].

OBJECTIVE: To investigate the role of visfatin in the pathogenesis of gestational diabetes mellitus (GDM) and its correlation with insulin resistance. METHODS: The study recruited 58 pregnant women of 24 to 28 gestational weeks in People's Hospital of Hebei Province from January to June 2013. Among them, 30 were patients with GDM (GDM group), 28 had normal oral glucose tolerance test and was referred as healthy pregnancy group (NGT group). Fourteen age-matched female who were first-degree relatives (FDR1) of type 2 diabetes mellitus patients, and 27 healthy nonpregnant women with normal oral glucose tolerance test were referred as high-risk group and normal controls (NC), respectively. The fasting plasma glucose (FPG), 1 hour and 2 hours postprandial glucose levels were measured by glucose oxidase method. The fasting insulin (FIN) levels were measured by radioimmunoassay and the homeostatic model assessment-insulin resistance index (HOMA- IR) was calculated. The levels of total cholesterol (TC), triglycerdes (TG), high density lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (LDL) were determined. The visfatin levels were measured by ELISA. RESULTS: (1)The levels of FPG were significantly higher in GDM, FDR1 and NC group [(5.5 ± 0.7), (5.1 ±0.6), (5.2 ± 0.4)mmol/L] than that in NGT group [(4.5 ± 0.3) mmol/L], respectively (P < 0.05). (2) The levels of INS [(14 ± 6)mU/L], HOMA- IR (4.0 ± 2.0), 1 hour [(10.9 ± 1.8) mmol/L] and 2 hours [(8.6 ± 1.8) mmol/L] postprandial glucose levels of GDM group were significantly higher than those in NGT group [(12 ± 4) mU/L, 2.0 ± 1.0, (7. 4 ± 1.3) and (6.2 ± 0.9) mmol/L], respectively (P < 0.05). (3) The levels of TC, TG, HDL and LDL levels in GDM group were (5.5 ± 0.9), (2.8 ± 0.8), (1.8 ± 0.4) and (3.3 ± 0.8) mmol/L, and were(5.9 ± 0.8), (2.5 ± 0.7), (1.9 ± 0.4) and (3.4 ± 0.6) mmol/L in NGT group. The levels of lipid in the two groups were significantly higher than those in FDR1 or NC group, respectively(P < 0.05).(4)The levels of visfatin in GDM group and NGT group [(43 ± 10), (45 ± 12) µg/L] were significantly higher than that in FDR1 or NC group [(29 ± 9), (36 ± 7) µg/L], respectively (P < 0.05), but the visfatin levels in FDR1 group were significantly lower than that in NC group (P < 0.05). The visfatin levels in GDM group were slightly lower than that in NGT group, but the difference was not statistically significant (P > 0.05). (5)The visfatin levels in NGT group were negatively correlated to the levels of FPG, HOMA-IR and TC (r = -0.38, -0.44, -0.47, respectively, P < 0.05). But the visfatin levels in GDM group were not correlated with the levels of FPG, HOMA-IR, TC (r = -0.16, -0.01, 0.33, respectively, P > 0.05). While in NC group, the levels of visfatin were negatively correlated with FPG and 2 hours postprandial glucose(r = -0.48, -0.42, respectively, P < 0.05). CONCLUSION: Visfatin may be an important adipokine that involved in the carbohydrate and lipid metabolism in GDM, and is related to the pathogensis of GDM and insulin resistance.

Serum Levels and Placental Expression of NGAL in Gestational Diabetes Mellitus.

OBJECTIVES: The aim was to investigate neutrophil gelatinase-associated lipocalin (NGAL) levels in the serum and term placentas and its potential role in gestational diabetes mellitus (GDM). METHODS: A total of 49 GDM subjects and 39 age-matched women with normal pregnancies were recruited. We examined serum concentrations of NGAL and tumor necrosis factor-α (TNF-α) in maternal blood and cord blood and their expression levels in the term placentas and umbilical cord. RESULTS: Serum NGAL levels were significantly higher in GDM patients than in normal pregnant controls both in the maternal blood (4.80 ± 1.99 vs. 3.66 ± 1.13, P=0.001) and the cord blood (4.70 ± 2.08 vs. 3.85 ± 1.44, P=0.027). Moreover, serum NGAL levels exhibited a positive correlation with various parameters of insulin resistance. Maternal serum NGAL levels positively correlated with the NGAL levels found in the cord blood of the control (r = 0.399, P=0.012) and the GDM subjects (r = 0.349, P=0.014). Finally, the expression of NGAL protein levels in the placenta (1.22 ± 0.39 vs. 0.65 ± 0.23, P < 0.001) and umbilical cord (0.65 ± 0.23 vs. 0.25 ± 0.10, P < 0.001) were higher in GDM women than those noted in the control subjects. In the GDM group, maternal serum NGAL levels exhibited a positive correlation with placental NGAL mRNA and protein levels (r = 0.848, P=0.008; r = 0.636, P=0.011, respectively). CONCLUSIONS: NGAL may be an important adipokine involved in GDM and fetal development. The oversecretion of NGAL from the placenta may contribute to the elevated levels of serum NGAL in gestational diabetes mellitus.

Placenta expression of vitamin D and related genes in pregnant women with gestational diabetes mellitus.

OBJECTIVES AND METHODS: A total of 41 GDM and 40 normal glucose tolerance subjects were recruited. Through detecting the level of Serum vitamin D with electrochemical luminescence and vitamin D receptor (VDR) with Enzyme-linked immunosorbent assay (ELISA) in maternal and cord blood, the expression leves of CYP24A1, CYP27B1, VDR protein and mRNA in placenta and umbilical cord with western blotting and RT-PCR, and the DNA methylation levels of CYP24A1 and CYP27B1 gene in placenta with methylation-specific PCR (MSP) and direct bisulfite sequencing (BSP) analysis to explore the potential role of the vitamin D and its related genes in gestational diabetes mellitus (GDM). RESULTS: Serum vitamin D concentrations were significantly higher in normal pregnant than women with GDM in maternal blood (P < 0.01) and cord blood (P = 0.014). Compared to the control group, the expression levels of CYP24A1 protein (P < 0.01) and mRNA (P = 0.021) and VDR protein (P = 0.026) and mRNA (P = 0.023) in the GDM group were significantly higher in placenta and umbilical cord tissues (P = 0.015, P < 0.01, P = 0.028, P < 0.01, respectively), while that of CYP27B1 protein (P < 0.01) and mRNA (P = 0.042) was significantly lower (P = 0.022, P = 0.032, respectively). Moreover, partial DNA methylation of CYP24A1 and CYP27B1 genes was observed in both GDM and control groups. CONCLUSIONS: Vitamin D deficiency participates in the pathogenesis of GDM, and changes in the expression of genes related to the vitamin D metabolic pathway are closely related to vitamin D levels in the pregnancy and fetus. However, DNA methylation of CYP24A1 and CYP27B1 might not be involved in the pathogenesis of GDM.

Visfatin is regulated by interleukin­6 and affected by the PPAR­Î³ pathway in BeWo cells.

Visfatin, an adipocytokine and cytosolic enzyme with nicotinamide phosphoribosyltransferase (Nampt) activity, is involved in the pathogenesis of numerous metabolic disorders. In addition, the nuclear receptor peroxisome proliferator­activated receptor­Î³ (PPAR­Î³) serves important roles in anti­inflammatory reactions and regulates glucose and lipid metabolism. The aim of the present study was to investigate the effect of interleukin­6 (IL­6) on the expression and secretion of visfatin in BeWo cells, and to determine whether the PPAR­Î³ pathway is involved in the regulation of visfatin by IL­6. Therefore, BeWo cells were stimulated with serial concentrations of IL­6 or pioglitazone, and the expression levels of visfatin and PPAR­Î³ were determined by reverse transcription­quantitative polymerase chain reaction and western blotting. The results of the present study demonstrated that IL­6 downregulated the mRNA levels of visfatin and PPAR­Î³, which were strongly associated. Activation of PPAR­Î³ by pioglitazone resulted in significantly increased expression of visfatin, which abrogated the inhibitory effect of IL­6 on visfatin in BeWo cells. Furthermore, treatment using pioglitazone alone increased the expression and secretion of the visfatin protein, compared with the control or IL­6 alone group. In summary, the findings of the present study suggested that IL­6 inhibited the expression of visfatin and PPAR­Î³ at the transcriptional level; in addition, activation of PPAR­Î³ upregulated visfatin at the mRNA and protein expression levels. Therefore, the PPAR­Î³ signaling pathway may be involved in the regulation of visfatin by IL­6 in BeWo cells. These results may provide novel insight into the roles of visfatin in trophoblastic cells. Furthermore, thiazolidinedione pioglitazone, by upregulating visfatin expression, may promote the energy metabolism of trophoblastic cells, maintain the function of the placenta and improve the outcome of pregnancy.