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Objective To evaluate the role of autophagy in hydrogen-induced inhibition of apoptosis in hippocampal neurons in a rat model of orthotopic liver transplantation (OLT).Methods Fifty-six pathogen-free healthy adult male Sprague-Dawley rats,aged 8-10 weeks,weighing 220-250 g,were used in the study.Thirty-two rats were selected and assigned into 4 groups (n =8 each) using a random number table:sham operation group (group S),OLT group,hydrogen-rich saline group (group HS) and chloroquine group (group CQ).The other 24 rats severed as the donors.In group S,laparotomy was performed,and the related blood vessels were isolated.The model of OLT was established in OLT,HS and CQ groups.In group OLT,normal saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group HS,hydrogen-rich saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group CQ,autophagy inhibitor chloroquine 60 mg/kg was injected intraperitoneally at 1 h before establishment of the model,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,the rats were sacrificed and hippocampi were isolated for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity,for pathological examination (with light microscope),and for detection of cell apoptosis (by TUNEL staining) and expression of autophagy-and apoptosis-related proteins caspase-3,cytochrome c (Cyt c),microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p53 in hippocampal tissues (by Western blot analysis).Apoptosis index (AI) was calculated.Results Compared with group S,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3,Cyt c,LC3 Ⅱ,Beclin-1 and p53 was up-regulated in OLT,HS and CQ groups (P＜0.05).Compared with group OLT,the MDA content and AI were significantly decreased,the SOD activity was increased,the expression of caspase-3 and Cyt c was down-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was up-regulated in group HS (P＜0.05).Compared with group HS,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3 and Cyt c was up-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was down-regulated in group CQ (P＜0.05).Conclusion The mechanism by which hydrogen inhibits apoptosis in hippocampal neurons is related to promotion of autophagy in a rat model of OLT.
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Objective To investigate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) / heme oxygenase-1 (HO-1) signaling pathway in reduction of acute kidney injury following orthotopic liver transplantation (OLT) by hydrogen-rich saline in rats.Methods Thirty-two healthy adult male SpragueDawley rats,weighing 220-250 g,were randomly assigned into 4 groups (n =8 each) using a random number table:sham operation group (S group),OLT group,hydrogen-rich saline group (HS group),and all-trans retinoic acid (ATRA) group.Laparotomy was performed,and the related blood vessels were isolated in group S.The model of orthotopic autologous liver transplantation was established in OLT,HS and ATRA groups.Normal saline and hydrogen-rich saline 6 ml/kg were injected through the inferior vena cava at 5 min before the portal vein was clamped in OLT and HS groups,respectively.In group ATRA,Nrf2 inhibitor ATRA 7 mg/kg was injected intraperitoneally once a day for 2 consecutive days,the model of orthotopic autologous liver transplantation was established at 16 h after the last injection of ATRA,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,blood samples were collected for determination of serum blood urea nitrogen (BUN),creatinine (Cr),interleukin10 (1L-10) and tumor necrosis factor-alpha (TNF-α) concentrations.After blood sampling,the lungs were removed for determination of malondialdehyde (MDA) content,superoxide dismutase (SOD) activity,expression of HO-1,Bcl-2 and Bax mRNA (by using real-time reverse transcriptase polymerase chain reaction),and HO-1 protein expression in lung tissues (by Western blot) and for microscopic examination.The damage to the renal tubules was scored.Results Compared with group S,the serum BUN,Cr and TNF-α concentrations were significantly increased,the serum IL-10 concentrations were decreased,the MDA content and renal tubular damage score were increased,the SOD activity was decreased,and the expression of HO-1 protein and mRNA,and Bcl-2 and Bax mRNA was up-regulated in group OLT (P＜ 0.05).Compared with group OLT,the serum BUN,Cr and TNF-α concentrations were significantly decreased,the serum IL-10 concentrations were increased,the MDA content and renal tubular damage score were decreased,the SOD activity was increased,the expression of HO-1 protein and mRNA and Bcl-2 mR-NA was up-regulated,and the expression of Bax mRNA was down-regulated in group HS (P＜0.05).Compared with group HS,the serum BUN,Cr and TNF-α concentrations were significantly increased,the serum IL-10 concentrations were decreased,the MDA content and renal tubular damage score were increased,the SOD activity was decreased,the expression of HO-1 protein and mRNA and Bcl-2 mRNA was down-regulated,and the expression of Bax mRNA was up-regulated in group ATRA (P＜0.05).Conclusion The mechanism by which hydrogen-rich saline reduces acute kidney injury following OLT is probably associated with activation of Nrf2/HO-1 signaling pathway in rats.
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Objective To evaluate the role of autophagy on acute kidney injury after liver transplantation.Method Fifty-six healthy male Sprague-Dawley rats were randomly assigned into 4 groups:sham group,orthotopic liver transplantation (OLT) group,sirolimus pretreated (SRL) group and 3-methyladenine pretreated(3-MA) group.OLT model was established.Then the rats were sacrificed at 6 h after reperfusion.The renal function and the extent of oxidative stress relative proteins malondialdehyde (MDA) and superoxide dismutase (SOD) were observed.The levels of apoptosis relative genes caspase-3 and cyt c and the expression of autophagy relative proteins were detected.The pathological changes were microscopically examined in renal tissues.TUNEL staining was used to observe the apoptosis of tubular epithelial cells.Transmission electron microscopy was applied to observe the ultrastructure changes of tubular epithelial cells.Result As compared with sham group,OLT and 3-MA groups showed a serious renal injury including cellular vacuolization,loss of brush borders,and a significant rise in BUN,Cr and MDA,while a decrease in SOD activity.The levels of caspase-3 mRNA and cyt c rnRNA were increased significantly.Whereas compared to OLT and 3-MA groups,renal function and oxidative stress levels in SRL group ameliorated,and histopathologic damage and apoptosis alleviated after OLT.Simultaneously,the levels of caspase-3 mRNA and cyt c mRNA were decreased.The expression of beclin-1 and LC3-]Ⅱ was effectively upregulated.Conclusion Autophagy could alleviate acute kidney injury after liver transplantation through inhibiting oxidative stress and apoptosis.
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<p><b>OBJECTIVE</b>To study the relationship between the TCM Syndrome Differentiation-types of congestive heart failure (CHF) and thyroid hormones, including triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH), and atrial natriuretic peptide (ANP), as well as cardiac function parameters, including left ventricular ejection fraction (LVEF), mean velocity of circumferentid fiber shortening (mVcf) and A peak/E peak (A/E).</p><p><b>METHODS</b>One hundred patients with CHF were divided into 4 Syndrome Differentiation-type groups, their cardiac function parameters, ANP and thyroid hormones were determined and compared with those in the 23 subjects in the control group.</p><p><b>RESULTS</b>In CHF patients with edema and blood stasis Syndrome type, the level of plasma ANP was significantly higher than that in the control group (P < 0.05); level of T3 was significantly lower than that in the control group and in CHF patients of other three (Xin-qi deficiency, Yin-deficiency and blood stasis) Syndrome groups (P < 0.01, P < 0.01, P < 0.05 and P < 0.01); levels of LVEF and mVcf were significantly lower than those in the other three Syndrome groups (all P < 0.01). Level of T4 in other three Syndrome groups significantly increased than that in the edema and blood stasis Syndrome type. A/E value showed a higher level in patients of all TCM type than that in the control (P < 0.01). Correlation analysis showed that T3 was positively correlated with LVEF and T4 (r = 0.200, P < 0.05, and r = 0.293, P < 0.01), and negatively correlated with ANP (r = -0.263, P < 0.01); T4 was negatively correlated with A/E (r = -0.226, P < 0.05).</p><p><b>CONCLUSION</b>The lowering of T3 and T4 and increasing of ANP may be one of the important reasons for lowering of LVEF in CHF patients with edema and blood stasis Syndrome-type. The decrease of T4 may be one of the important reasons for elevation of A/E and aggravation of left ventricular diastolic dysfunction in CHF patients of all the 4 TCM Syndrome-types.</p>