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Objective To explore the influences of plumbagin on phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (Akt) signaling pathway in rats with carbon tetrachloride induced liver fibrosis.Methods Forty male SD rats were assigned to control group,model group,2 mg/kg plumbagin treated group and 3 mg/kg plumbagin treated group,with 10 rats in each group.Rats of the control group were injected with 0.9％ NaCl solution (2 mL/kg) intraperitoneally,while rats of the other three groups were injected with 60％ carbon tetrachloride/peanut oil (2 mL/kg,3 times a week for 6 weeks) intraperitoneally.Since the third week after modeling,rats of plumbagin treated groups were treated with intraperitoneal injection of plumbagin at a dose of 2 mg/kg and 3 mg/kg (twice a week for 4 weeks),respectively.Serum levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),albumin (Alb) were monitored routinely.Hyaluronic acid (HA) and laminin (LN) were measured by radioimmunoassay after 6 weeks; protein expression of liver PI3K,Akt and phosphorylated Akt (p-Akt) were evaluated by Western blotting and immunohistochemistry,respectively.Comparison of means among groups was performed by univariate analysis of variance.Results The hepatic fibrosis model was successfully established after 6 weeks.Serum levels of ALT,AST,HA and LN of model group were significantly higher than those of control group (all P＜0.05).Serum levels of ALT,AST,HA and LN of 2 mg/kg plumbagin treated group and 3 mg/kg plumbagin treated group were significantly lower than those of model group,and the differences were statistically significant (all P＜0.05).The protein expressions of PI3K and Akt in each group were comparable (all P＞0.05).The p-Akt protein was mainly expressed in nucleus of hepatocytes.The levels of p-Akt protein in control group,model group,2 mg/kg plumbagin treated group and 3 mg/kg plumbagin treated group were 0.0821±0.0003,0.7374±0.0037,0.3679 ±0.0332 and 0.1327±0.0561,respectively,and that in model group was significantly higher than control group (t =851.302,P＜0.05),but those in plumbagin treated groups were both lower than model group (t=71.858 and 28.363,all P＜0.05).Conclusions Plumbagin presents anti-fibrotic effects in the liver,by down-regulating the expression of p-Akt during the development of fibrosis,which might be one of the antifibrotic mechanisms.
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ObjectiveTo investigate the effects of 5(S),6(R),7-trihydroxyheptanoic acid methyl ester (BML-111) on pregnant mice with fetal growth restriction(FGR) induced by antenatal dexamethasone and its probable mechanism. MethodsThe mice were mated overnight,with day 1 of pregnancy designated as the day on which spermatozoa were presented in a vaginal smear.The pregnant mice were then randomly divided into control group,dexamethasone group and BML-111 group.From 9 to 14 days of pregnancy,pregnant ICR mice of control,dexamethasone and BML-111 group were treated separately with saline,dexamethasone(5 mg/kg) and dexamethasone at 8:00 am,and two hours later they were treated separately again with 1 mg/kg saline,saline and BML-111.On the day 18 of gestation,they were sacrificed after blood were collected from their eyeballs.The serum lipoxin A4 was measured with enzyme-linked immunosorbent assay. Fetuses were delivered by cesarean section; the placenta and uterus were immediately removed and frozen.Gene expressions of 11β-hydroxysteroid dehydrogenase 2 ( 11β-HSD2 ),interleukin-1β (IL-1β) in placenta and lipoxin A4 receptor-formyl peptide receptor 3 (FPR3)in uterine were detected by reverse transcriptionpolymerase chain reaction and compared with analysis of variance.The 11β-HSD2 protein in mice placenta was detected by immunohistochemistry. ResultsThe mean fetal weight of dexamethasone group was (0.823±0.054) g,lower than that of the control group and BML-111 group [(1.103±0.218) g and (0.992 ± 0.207) g] (t =- 4.108 and - 2.890,P ＜ 0.05 respectively).Protein expression of 11β-HSD2 in dexamethasone group (0.030±0.019) was weaker than that in control group (0.058±0.015,t=-3.107,P＜0.05) or in BML-111 group (0.049±0.026,t=-2.211,P＜0.05).The expression of 11β-HSD2 mRNA in dexamethasone group (0.457±0.062) was lower than that in control group (0.943±0.057,t=-9.418,P＜0.05) or in BML-111 group (0.698±0.071,t=-4.617,P＜0.05).Expression of IL-1β mRNA in dexamethasone group (0.543±0.103)was less than that in control group (0.710± 0.085,t=-3.736,P＜0.05) but more than that in BML-111 group (0.229 ±0.031,t=7.025,P＜0.05). The expression of FPR3 mRNA in dexamethasone group (0.323 ± 0.019) was less than that in control group (0.857 ± 0.057,t =-14.630,P＜0.05) or in BML-111 group (0.499 ±0.050,t=-4.822,P＜0.05).The serum concentration of lipoxin A4 in dexamethasone group was lower than that in control group [(64.463±22.144) pg/ml vs (101.610±24.916) pg/ml,t=3.152,P＜0.05].ConclusionsBML-111 regulate the expression of 11β-HSD2 and then protect against FGR resulted from too much prenatal application of dexamethasone.
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Objective To investigate the expression and clinical significance of lipoxin A4,leukotrienc C4,lipoxygenase-5 in peripheral blood of pregnant women with different types of severe preeclampsia.Methods Forty-five singleton pregnant women who accepted antenatal care and delivered in First Affiliated Hospital of Wenzhou Medical College were enrolled in this study from December 2010 to June 2011.All objects were divided into normal pregnancy group (n=20),early onset severe preeclampsia group (n=10) and late onset severe preeclampsia group (n=15).Enzymelinked immunosorbent assay was used to detect lipoxin A4 and leukotriene C4 levels in peripheral blood.The level of lipoxygenase-5 mRNA in white blood cells was detected by real time fluorescence quantitative reverse transcription-polymerase chain reaction.The differences of lipoxin A4,leukotriene C4 and lipoxygenase-5 mRNA among groups were compared by analysis of variance and LSD-t test;and correlations among their expressions were analyzed by linear regression.Results Lipoxin A4 level in early and late onset severe preeclampsia group was (355.3±116.0) pg/ml and (389.7±117.5) pg/ml,which were both significantly lower than that in normal pregnancy group [(555.0±139.8) pg/ml] (t=-4.03 and-3.77,P＜0.05 respectively).The leukotriene C4 level in early and lateonset severe preeclampsia and normal pregnaney group was (591.3±185.5) pg/ml,(510.3±197.1) pg/ml and (496.9 ± 158.8) pg/ml,no statistical difference were found (F=0.889,P＞0.05) ; neither did the expression of lipoxygenase 5 mRNA,which was 4.2± 1.9 in normal pregnancy group,4.8 ± 2.0 in early onset severe preeclampsia group and 4.4 ± 1.2 in late onset severe preeclampsia group (F=0.311,P＞0.05).There was no correlation among the levels of lipoxin A4,leukotriene C4 and lipoxygenase-5 mRNA in each group (P ＞ 0.05).Conclusions Early and remarkable decreasing of lipoxin A4 level might contribute to the development of early onset severe preeclampsia.
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Objective To explore lipoxinA4 (LXA4) expression in maternal serum of pregnant women and the protective effect and mechanism of LXA4 on trophoblastic cells from oxidative injury. Methods Trophoblastic cells were randomized into six groups: Control group; Lipopolysaccharides (LPS) group, cells were stimulated by 10 μg/ml LPS for 24 h; Intervention group, cells stimulated by LPS were treated with 100 nmol/L LXA4 for 24 h; LXA4 group, cells were treated with 100 nmol/L LXA4 for 24 h; Antagonistic group, cells stimulated by LPS were treated with 100 nmol/L LXA4 plus 100 μmol/L N-tert-butoxycarbonyl-2-pyrrolidine (BOC-2) for 24 h; BOC-2 group, trophoblastic cells stimulated by LPS were treated with 100 μmol/L BOC-2 for 24 h. The serum concentration of LXA4 in normal group and preeclampsia group was detected by ELISA. The intracellular formation of reactive oxygen species (ROS) was detected by 2,7-dichlorofluorescein diacetate (DCFH-DA) as a fluorescent probe. SOD mRNA was analyzed by RT-PCR. SOD and Nrf2 protein expressions were analyzed by Western blot. The levels of SOD in trophoblastic cells were detected by using detection kit. Results (1) The serum concentration of LXA4 was significantly lower in preeclampsia group (165.53±18.89) pg/L than in the control [(545.67±30.91) pg/L, P0.05). Conclusions LXA4 can significantly reduce the oxidative stress of placental trophoblastic cells stimulated by LPS. LXA4 can bind to lipoxin receptors and activate Nrf2-ARE signaling pathway playing a protective effect. So LXA4 in pregnant women can affect the oxidative stress of placenta.
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Objective To observe the effect of shock lymph drainage on multiple organ injury of rats with traumatic hemorrhagic shock (THS) and discuss the relating mechanism. Methods Male Wistar rats were divided into control group, lymph drainage group and lymph return group. The THS model was established in lymph drainage group and lymph return group, when the shock mesenteric lymph was drained in lymph drainage group. The change of the mean arterial pressure ( MAP), the biochemical indices of liver, kidney, myocardium and acid-base, the morphology, ATP contents and ATPase activities of lung, kidney, liver and myocardium were observed. Results The MAP at multiple time points after 80 minutes of infusion, the ATP contents and ATPase activities of multiple organs in lymph drainage group were higher than those in lymph return group. Multiple biochemical indices in lymph drainage group were superior to those in lymph return group, with statistical difference. The inflammation, congestion, degeneration and necrosis were found in organs of lymph return group, but only mild lesions could be seen in lymph drainage group. Conclusions The shock lymph drainage can alleviate multiple organ injury of THS rats, mechanism of which is correlated with improvement of the energy metabolism and maintenance of MAP and acid-base status.