Objective To investigate the effect of adiponectin on proliferation of airway smooth muscle cells( ASMCs) , and explore its possible mechanism. Methods ASMCs were derived fromrat airway tissue and were cultured in vitro. RT-PCR was used to verify the expression of adiponectin receptors on ASMCs. Then ASMCs were treated with adiponectin at different concentrations( 5, 10, 20, 40, 80 μg/mL) for different periods of time( 1, 12, 24, 48, 72 hours) , respectively. The absorbsence ratios of adiponectin at different concentrations were determined by MTT assay. The adenosine monophosphate-activated protein kinase( AMPK) and phosphorylated AMPK( pho-AMPK) in ASMCs were quantified by Western blot after being treated with adiponectin at different concentrations ( 5, 10, 20, 40 μg/mL) for 48 hours. ResultsThe inhibition of adiponectin on ASMCs was showed in dose-dependent manner( r = 0. 324, P lt; 0. 01) and time-dependent manner( r = 0. 607, P lt; 0. 05) . Western blot indicated that the expression of pho-AMPK increased with the increased concentrations of adiponectin( r =0. 607, P lt; 0. 01) . The ratio of pho-AMPK/AMPK were ( 27. 66 ±1. 03) % , ( 31. 91 ±0. 86 ) %, ( 75. 52 ±2. 67) % , and ( 84. 50 ±1. 05) % ,respectively, with significant differences between each concentrations of adiponectin( P lt; 0. 05) . There was no expression of pho-AMPK in the control group. Conclusion Adiponectin can significantly inhibit ASMCs’proliferation by activating AMPK.
Objective To investigate the effects of sodium valproate (VPA) in inhibiting Erastin-induced ferroptosis in bone marrow mesenchymal stem cells (BMSCs) and its underlying mechanisms. Methods BMSCs were isolated from bone marrow of 8-week-old Spragur Dawley rats and identified [cell surface antigens CD90, CD44, and CD45 were analyzed by flow cytometry, and osteogenic and adipogenic differentiation abilities were assessed by alizarin red S (ARS) and oil red O staining, respectively]. Cells of passage 3 were used for the Erastin-induced ferroptosis model, with different concentrations of VPA for intervention. The optimal drug concentration was determined using the cell counting kit 8 assay. The experiment was divided into 4 groups: group A, cells were cultured in osteogenic induction medium for 24 hours; group B, cells were cultured in osteogenic induction medium containing optimal concentration Erastin for 24 hours; group C, cells were cultured in osteogenic induction medium containing optimal concentration Erastin and VPA for 24 hours; group D, cells were cultured in osteogenic induction medium containing optimal concentration Erastin and VPA, and 8 μmol/L EX527 for 24 hours. The mitochondrial state of the cells was evaluated, including the levels of malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species (ROS). Osteogenic capacity was assessed by alkaline phosphatase (ALP) activity and ARS staining. Western blot analysis was performed to detect the expressions of osteogenic-related proteins [Runt-related transcription factor 2 (RUNX2) and osteopontin (OPN)], ferroptosis-related proteins [glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), and solute carrier family 7 member 11 (SLC7A11)], and pathway-related proteins [adenosine monophosphate-activated protein kinase (AMPK) and Sirtuin 1 (SIRT1)]. Results The cultured cells were identified as BMSCs. VPA inhibited Erastin-induced ferroptosis and the decline of osteogenic ability in BMSCs, acting through the activation of the AMPK/SIRT1 pathway. VPA significantly reduced the levels of ROS and MDA in Erastin-treated BMSCs and significantly increased GSH levels. Additionally, the expression levels of ferroptosis-related proteins (GPX4, FTH1, and SLC7A11) significantly decreased. VPA also upregulated the expressions of osteogenic-related proteins (RUNX2 and OPN), enhanced mineralization and osteogenic differentiation, and increased the expressions of pathway-related proteins (AMPK and SIRT1). These effects could be reversed by the SIRT1 inhibitor EX527. ConclusionVPA inhibits ferroptosis in BMSCs through the AMPK/SIRT1 axis and promotes osteogenesis.
Objective
To evaluate the inhibiting effect of adenosine on rat retinal ganglion cells (RGC) death induced by P2X7 and N-methyl-D-aspartate (NMDA) receptor.
Methods
(1) Long-Evan neonatal rats were back labeled with aminostilbamidine to identify RGC. The viability of RGC affected by P2X7 excitomotor BzATP (50 mu;mol/L), glutamate receptor excitomotor NMDA (100 mu;mol/L) and adenosine (300 mu;mol/L) was detected. (2) RGC from the retinae of unlabeled neonatal rats were cultured in vitro. After labeled with Fura-2 methyl acetate, an intracellular calcium indicator, the effect of BzATP, NMDA and adenosine on intracellular Ca2+ level was detected byCa2+ imaging system.
Results
Both BzATP (50 mu;mol/L) and NMDA(100 mu;mol/L) could kill about 30% of the RGC. Cell death was prevented by adenosine (300 mu;mol/L) with the cell viability increased from (68.9plusmn;2.3)% and (69.9plusmn;3.2)% to (91.2plusmn;3.5)% (P＜0.001) and (102.1plusmn;3.9)% (P＜0.001), respectively. BzATP (50 mu;mol/L) led to a large, sustained increase of intracellular Ca2+ concentration to (1183plusmn;109) nmol/L. After the adenosine intervened, Ca2+ concentration increased slightly to (314plusmn;64) nmol/L (P＜0.001).
Conclusion
Adenosine may prevent RGC death and increase of intracellular Ca2+ concentration from P2X7and NMDA receptor stimulation.
(Chin J Ocul Fundus Dis, 2007, 23: 133-136)
The aim of this study is to assess ischemia/reperfusion injury in carbon tetrachloride induced cirrhotic liver as compared to normal liver in the rats. Results showed that in cirrhotic liver, instead of diminishing the hepatic vein nitric oxide level increased significantly after ischemia from 8.04 μmol/L to 11.52 μmol/L and remained high till 5 hrs after reperfusion. The hepatic adenosine triphosphate (ATP) contents decreased as that seen in normal rat but did not restore to normal till the end of 5 hrs after reperfusion. Based on these findings, it is postulated that in cirrhotic liver, ischemia/reperfusion injury is aggrvated as evidenced by of nitric oxide, and extended diminishing in ATP.
ObjectiveTo explore the clinical value of age/pleural fluid adenosine deaminase (age/ADA) ratio and serum lactate dehydrogenase/pleural fluid adenosine deaminase ratio (Cancer Ratio, CR) in the diagnosis of malignant pleural effusions (MPE). MethodsThe study collected 44 patients with MPE and 48 patients with benign pleural effusion (BPE) to compare the differences in age, gender, carcinoembryonic antigen (CEA), age/ADA ratio and CR between the groups. The receiver operating characteristic (ROC) curve of CEA, age/ADA and CR was constructed and the area under the ROC curve (AUC), sensitivity and specificity was calculated to identify the diagnostic performance of the three indicators alone or in combination in MPE. ResultsCEA, age/ADA and CR were significant higher in the MPE group than those in the BPE group (all P<0.05), the AUCs of CEA, age/ADA and CR were 0.768, 0.837 and 0.866, respectively; the sensitivity was 61.36%, 88.64% and 81.82%, the specificity was 85.42%, 75.00%, 83.33%, respectively. The AUCs of CEA combined with age/ADA, CEA combined with CR, age/ADA combined with CR, CEA combined with age/ADA and CR were respectively 0.892, 0.911, 0.837 and 0.907; the sensitivity was 81.82%, 86.36%, 88.64% and 90.91%, the specificity was 79.17%, 79.17%, 75.00% and 77.08%, respectively. ConclusionsAge/ADA and CR demonstrated good diagnostic performance in MPE, moreover, the diagnostic performance can be further improved when combined with the traditional tumor marker CEA, and more research about its diagnostic value is needed in the future.
Objective To investigate the effects of adenosine 2A receptor (A2AR) activation on oxidative stress in small-forsize liver transplantation. Methods A rat orthotopic liver transplantation model was performed using 40% graft, 18 recipients were given intravenously saline (control group), CGS21680 (A2AR agonist, CGS21680 group) or ZM241385 (A2AR antagonist, CGS21680+ZM241385 group) randomly. Aspartate aminotransferase (AST), enzymatic antioxidants 〔superoxide dismutase (SOD); catalase (CAT); glutathione peroxidase (GSH-Px)〕, non-enzymatic antioxidants 〔ascorbic acid (AA); glutathione (GSH); α-tocopherol (TOC)〕 and lipid oxidant metabolites malondialdehyde (MDA) were measured and analyzed at 6 h after reperfusion. Results Compared with the control group and CGS21680+ZM241385 group, A2AR activation increased the activities of SOD and GSHPx (Plt;0.05), reduced the productions of AST and MDA (Plt;0.05), increased the levels of AA, GSH and TOC (Plt;0.05) in CGS21680 group. But there was no significant change in CAT activity (Pgt;0.05) among 3 groups. Conclusions A2AR activation improves the antioxidant enzyme activities, promotes the production of antioxidants, and slowes down the increase in MDA level, depresses of the increase in AST activity. A2AR activation suppresses oxidative damage and increases the antioxidant capacity which in turn minimizes their harmful effects of ischemia-reperfusion in small-for-size liver transplantation.
ObjectiveTo systematically review the clinical efficacy and effects on pregnancy outcomes of S-adenosy-L-methionine combined with ursodesoxycholic acid in the treatment of intrahepatic cholestasis of pregnancy.
MethodsDatabases such as PubMed, The Cochrane Library, CNKI, VIP, WanFang Data were searched for the studies about the clinical efficacy and effects on pregnancy outcomes of S-adenosy-L-methionine combined with ursodesoxycholic acid in the treatment of intrahepatic cholestasis of pregnancy up to December 31st, 2013. Two reviewers independently screened literature, extracted data and evaluated methodological quality. Then meta-analysis was conducted using RevMan 5.0.24 software.
ResultsA total of 11 RCTs involving 776 patients were included. The results of meta-analysis showed that, combined medication reduced blood biochemical indexes inlcuding ALT (MD=3.63, 95%CI 0.63 to 6.64, P=0.02), TB (MD=3.70, 95%CI 1.45 to 5.96, P=0.001), and AST (MD=7.61, 95%CI 2.47 to 12.75, P=0.004). Combined therapy significantly decreased the rates of amniotic fluid contamination (OR=0.29, 95%CI 0.19 to 0.45, P=0.000 01), cesarean section (OR=0.53, 95%CI 0.36 to 0.79, P=0.002), postpartum hemorrhage (OR=0.32, 95%CI 0.12 to 0.90, P=0.03), preterm birth (OR=0.36, 95%CI 0.24 to 0.55, P < 0.000 01), fetal distress (OR=0.33, 95%CI 0.19 to 0.58, P=0.000 1) and neonates asphyxia (OR=0.30, 95%CI 0.19 to 0.47, P < 0.000 01). Combined therapy was also beneficial to improving pruritus symptoms (MD=0.20, 95%CI 0.08 to 0.31, P=0.000 08) and benefiting fetus growth (MD=0.45, 95%CI 0.23 to 0.66, P < 0.000 1).
ConclusionThe combination of S-adenosy-L-methionine and ursodesoxycholic acid is superior to ursodesoxycholic acid alone in improving clinical symptoms and pregnant outcomes of patients with intrahepatic cholestasis of pregnancy.
The Dacron grafts seeded with autologous venous fragments were implanted into IVC of 13 canines as seeded group and the control grafts (8 cases), which were only preclotted with fresh blood. The amounts of cAMP and cGMP in serum and within platelet were measured. All of the specimens explanted at exsaguination were observed morphologically. The results shown that the total patency rate were 61.5% in seeded group, but 25.0% in control one and new endothelial lining formed at two weeks after implantation of the seeded grafts. The amounts of cAMP in serum and within platelet were higher in seeded group, but the amounts of cGMP were lower in serum and within platelet. These were in accordance with the results that the endothelialization of the grafts were complete in seeded group but not complete in control one. The results indicate that seeding Dacron with autologous venous fragment makes new endothelium formed at two weeks after implantation, increases the amounts of cAMP in serum and within platelet, but reduces the amounts of cGMP and thus improves graft patency rate.
