ABSTRACT
Objective:To observe the effect of sodium fluoride (NaF) on the expression of phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signal and apoptosis in human osteosarcoma Saos-2 cells.Methods:Saos-2 cells were divided into 0 (control), 5, 10, 20 and 40 mg/L fluoride groups ( n= 3) according to the dose of NaF. The cells were collected after 24 and 48 h of culture in vitro. Western blotting was used to detect the expressions of PI3K/Akt signal and Forkhead transcription factor (FoxO) 1, and the apoptosis level of Saos-2 cells was detected by flow cytometry. Results:There were no significants differences in the expressions of PI3K and Akt in Saos-2 cells at 24 and 48 h ( P > 0.05). At 24 h, the expressions of phosphorylated PI3K (p-PI3K) in 5, 10, 20 and 40 mg/L fluoride groups (0.40 ± 0.06, 0.45 ± 0.02, 0.37 ± 0.06, 0.32 ± 0.06) were higher than that in the control group (0.28 ± 0.04, P < 0.05); at 48 h, the expressions of p-PI3K in 5 and 10 mg/L fluoride groups (0.46 ± 0.06, 0.58 ± 0.03) were higher than that in the control group (0.29 ± 0.04, P < 0.05), and that in the 40 mg/L fluoride group (0.21 ± 0.03) was lower than that in the control group ( P < 0.05). At 24 h, the expressions of phosphorylated Akt (p-Akt) in 5, 10 and 20 mg/L fluoride groups (0.27 ± 0.01, 0.30 ± 0.03, 0.27 ± 0.03) were higher than that in the control group (0.20 ± 0.02, P < 0.05); at 48 h, the expressions of p-Akt in 5 and 10 mg/L fluoride groups (0.42 ± 0.04, 0.60 ± 0.02) were higher than that in the control group (0.27 ± 0.01, P < 0.05), and that in the 40 mg/L fluoride group (0.18 ± 0.02) was lower than that in the control group ( P < 0.05). The expressions of FoxO1 in 10, 20 and 40 mg/L fluoride groups at 24 h (0.38 ± 0.07, 0.41 ± 0.06, 0.47 ± 0.08) were higher than that in the control group (0.34 ± 0.04, P < 0.05). At 48 h, the expressions of FoxO1 in 5, 10, 20 and 40 mg/L fluoride groups (0.36 ± 0.08, 0.37 ± 0.10, 0.54 ± 0.04, 0.73 ± 0.04) were higher than that in the control group (0.28 ± 0.04, P < 0.05). At 24 and 48 h, the apoptosis rates of control group and 5, 10, 20 and 40 mg/L fluoride groups were (2.867 ± 0.583)%, (3.647 ± 0.035)%, (3.773 ± 0.095)%, (5.440 ± 0.325)%, (7.203 ± 0.476)%; (3.707 ± 0.286)%, (4.023 ± 0.241)%, (4.970 ± 0.368)%, (12.473 ± 0.949)%, (19.387 ± 1.892)%, respectively. The apoptosis level of 40 mg/L fluoride group was higher than that of control group at 24 h ( P < 0.05), and that of 20 and 40 mg/L fluoride groups were higher than that of control group at 48 h ( P < 0.05). Conclusion:Fluoride can directly activate the PI3K/Akt signaling pathway in osteoblasts, and then has anti-apoptotic effect.
ABSTRACT
Objective To investigate the expression of growth arrest-specific protein 6 (Gas6) in the placenta and decidua tissues and its relationship with the pathogenesis of preeclampsia. Methods All the patients were recruited in Qilu Hospital of Shangdong University from October 2013 to June 2014. Among them, thirty-two women with early-onset severe preeclampsia who received cesarean section were assigned to the preeclampsia group, and thirty healthy pregnant women who received cesarean section were defined as the control group. Blood glucose, blood lipids, platelet count, D-dimer levels and other clinical indicators of the two groups were detected. Immunohistochemistry of SP was conducted to identify the localization of Gas6 protein in the placenta and decidua tissues. And reverse transcription (RT)-PCR was performed for quantitative analysis of Gas6 RNA expression in placentas. The correlations between placental Gas6 mRNA levels with clinical indicators were analyzed. Results (1) The gestational age at delivery, blood pressure, serum albumin, platelet count and birth weight of fetuses showed statistically significant differences between the two groups(P<0.05). (2) The Gas6 protein expressed in the cytoplasm and nucleus of the syncytiotrophoblasts and decidual cells in the placenta and decidual tissues of the two groups. (3) The Gas6 mRNA expression elevated significantly in the placenta of preeclampsia group (0.60 ± 0.38) when compared to that of the control group (0.34±0.22;P<0.05). (4) The expression of Gas6 mRNA was positively related with body mass index, diastolic blood pressure, systolic blood pressure, free fatty acids and creatinine (P<0.05), while it was negatively associated with serum albumin(P<0.05). Conclusion The high expression of Gas6 in the placenta and decidua tissues may be related to the pathogenesis of severe preeclampsia.
ABSTRACT
Objective To investigate the expression of interleukin-37 (IL-37) in placenta tissue and its relationship with the pathogenesis of severe preeclampsia. Methods All the patients were recruited in Qilu Hospital of Shangdong University from November 2012 to November 2013. Among them, thirty patients with severe preeclampsia were assigned to the preeclampsia group, and thirty-one healthy pregnant women were assigned to the control group. Immunohistochemistry of SP was used to detect the IL-37 protein expression in placenta tissue of the two groups. The expression level of IL-37 in placenta tissue of the two groups was detected by western blot. Besides, reverse transcription(RT)-PCR was used to detect the expression of IL-37 mRNA. The correlation between the expression of IL-37 mRNA and the delivery gestational age, body mass index(BMI)was analyzed. Results (1) IL-37 were detected in the placenta of both the preeclampsia group and the control group, and the expression site mainly located in the syncytiotrophoblast, with a small amount in cytotrophoblast.(2)The expression levels of IL-37 protein in the preeclampsia group and the control group were 0.59 ± 0.39 and 0.88 ± 0.22, respectively. The IL-37 mRNA levels in the preeclampsia group and the control group were 0.55 ± 0.17 and 1.11 ± 0.21, respectively. Both decreased significantly when compared to the control group (P0.05) was seen neither in the preeclampsia group nor in the control group(r=-0.053, P>0.05). In the severe preeclampsia group, the pregnant women′s BMI had no significant correlation with IL-37 mRNA expression of placenta tissue(r=0.102, P>0.05), neither did the control group(r=-0.115, P>0.05). Conclusion IL-37 expression is significantly lower in severe preeclampsia placenta tissue than that in the normal pregnant women, which may play a protective role in the course of severe preeclampsia.