Objective To investigate the effect of S-adenosylmethionine (SAM) on mitochondrial injury that was induced by ischemia-reperfusion in rat liver. Methods Fifty-four rats were randomly divided equally into 3 groups: control group, ischemia-reperfusion group (I/R group), and SAM-treated group (SAM group). Hepatic ischemia had been only lasted for 30 min by obstructing the blood stream of hepatic portal vena (the portal vena was only separated but not obstructed in control group). The rats of SAM group received SAM intraperitoneally 2 h prior to ischemia. Blood samples of each group were collected from the inferior cava vena at 0, 1 and 6 h after reperfusion and the serum levels of AST and ALT were detected. Mitochondrial super oxidedismutase (SOD), malondialdehyde (MDA), adenosine triphosphate (ATP) and energy charge (EC) in samples of liver tissue were detected, and the mitochondrial ultrastructure was observed with electronmicroscope. Results The serum levels of AST, ALT and mitochondrial MDA at 0, 1 and 6 h after reperfusion in the I/R group were significantly higher than those in the control group, whereas the levels of mitochondrial SOD, ATP and EC were significantly lower than those in the control group (P＜0.01). Except the value of 0 h, when it comes to SAM group, the levels of AST, ALT and mitochondrial MDA were significantly lower (P＜0.05) and the levels of mitochondrial SOD, ATP and EC were significantly higher (P＜0.05, P＜0.01) than those in the I/R group, respectively. The mitochondrial ultrastructure was injured obviously in I/R group when compared with that in control group. The number of mitochondria decreased and the mitochondria swelled, making the crista became obscure and the density of matrix became lower. The above changes in SAM group were less obvious when compared with those in I/R group. Conclusion SAM may protect mitochondrion against hepatic ischemia injury, since it may prevent mitochondrial lipid peroxidation, increase ATP, and eventually improve energy metabolism after ischemia-reperfusion.
Objective To investigate the effect of S-adenosyl-l-methionine (SAM) on oxidative stress and alveolar septal cell apoptosis in mice with emphysema after smoking cessation. Methods Twenty-two male SPF C57BL/6J mice aged 6 - 8 weeks were randomly divided into 4 groups, ie. a healthy control group, an emphysema group, a smoking cessation group, and a SAM intervention for 8 weeks after smoking cessation group, with 8 mice in each group. The mice model of emphysema was established by intraperitoneal injection of cigarette smoke extract (CSE) combined with cigarette smoke exposure. Smoking cessation started after the emphysema model was successfully constructed and lasted for 8 weeks. After smoking cessation, the mice in SAM intervention groups were intraperitoneally injected with SAM mg·kg–1·d–1 for 8 weeks. The right lung sections of the mice were taken for hematoxylin-eosin staining to observe pathological changes, and the mean linea rintercept (MLI) and mean alveola rnumber (MAN) of lungs were measured. The concentrations of malondialdehyde (MDA), superoxide-dismutase (SOD) and glutathione (GSH) in alveolar lavage fluid of left lung were detected by spectrophotometry. Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) technique was carried out to detect the alveolar septal cells apoptosis. Results MLI, apoptosis index of alveolar septal cell and MDA concentration in bronchoalveolar lavage fluid (BALF) increased significantly in the emphysema group compared with healthy controls, increased significantly in the smoking cessation group compared with the emphysema group, and decreased in the SAM intervention group compared with the smoking cessation group (all P<0.05). GSH concentration and SOD activity in BALF and MAN was significantly lower in the emphysema group compared with the healthy control group, significantly lower in the smoking cessation group compared with the emphysema group, and significantly higher in the SAM intervention group compared with the smoking cessation group (all P<0.05). Conclusions Lung histopathology and apoptosis of alveolar septal cells in emphysema mice progress continuously after smoking cessation. SAM may reduce oxidative stress and improve apoptosis of alveolar septal cells, so as to protect emphysema mice after smoking cessation.
ObjectiveTo investigate the effects of overexpression of alpha/beta hydrolase domain-containing protein 5 (ABHD5) on the invasion and migration of human colon cancer cell line HCT116 and the pathway of adenosine monophosphate-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR).MethodsThe expression of ABHD5 in colon cancer tissues and its relationship with clinicopathological features was analyzed by UALCAN database. HCT116 cells were cultured in vitro and transfected with ABHD5 recombinant plasmid, then they were divided into control group, negative transfection group and ABHD5 transfection group. Real time quantitative PCR (qRT-PCR) was used to detect the expression of ABHD5 mRNA in HCT116 cells. The proliferation of HCT116 cells was detected by CCK-8 method. Transwell assay was used to detect the invasion and migration of HCT116 cells. The expression of matrix metalloprotein 9 (MMP-9), E-cadherin, Snail, and AMPK/mTOR pathway proteins p-AMPK, AMPK, p-mTOR and mTOR were detected by Western blot.ResultsThe results of the UALCAN showed that compared with normal colon tissues, the expression of ABHD5 mRNA in colon cancer tissues was decreased (P<0.05), and which in the adenocarcinoma and the N1 stage was lower than that of the mucinous adenocarcinoma (P<0.05) and N0 stage (P<0.05), respectively. Compared with the control group and the negative transfection group, the expression of ABHD5 mRNA in the ABHD5 transfection group was increased (P<0.05), the proliferation inhibition rate of HCT116 cells in the ABHD5 transfection group was increased (P<0.05), the numbers of migration and invasion cells in the ABHD5 transfection group were decreased (P<0.05), the expressions of MMP-9, Snail, p-mTOR and mTOR were reduced, and the expressions of E-cadherin, p-AMPK and AMPK were increased (P<0.05).ConclusionsThe overexpression of ABHD5 can inhibit the invasion and migration of colon cancer HCT116 cells, activate AMPK, and inhibit the expression of mTOR. It suggests that ABHD5 may play a role in inhibiting colon cancer by affecting AMPK/mTOR pathway.
目的:评价国产腺苷临床应用安全性及腺苷负荷心肌灌注显像对冠心病的诊断价值。方法:对51例临床疑诊冠心病患者,分别静脉注射腺苷,注射3min末,静脉注射核素显像剂99TcmMIBI 925 MBq,1～1.5h行心肌灌注断层显像,分析患者在腺苷注入后的血流动力学改变、副作用发生情况。经半年以上随访排除或确诊冠心病,评价腺苷负荷心肌灌注显像对冠心病的诊断价值。结果:腺苷输注后,86.3%(44/51) 患者出现各种副作用,停药后均很快缓解。腺苷负荷试验心肌灌注显像诊断冠心病的敏感性90.9%,特异性71.4%,准确性88.2%,阳性预测值95.2%,阴性预测值55.6%。结论:国产腺苷可安全地应用于负荷心肌灌注显像。腺苷负荷心肌灌注显像诊断冠心病敏感性较高,具有重要的临床应用价值。
Objective To investigate the protective effects of ischemic postconditioning (IPo) on ischemiareperfusion (I/R) myocardium and the relationship with mitochondrial adenosine triphosphate (ATP) sensitive K+ channels (mitoKATP) and provide evidences to the development of druginduced postconditioning. Methods Langendorff models were established in 40 Wistar rats which were divided into 5 groups by random number table with 8 rats in each group. Normal control group(NC group): the rat hearts were continuously reperfused by KrebsHenseleit bicarbonate buffer (K-HB) for 100 min without any other treatment; I/R group: the rat hearts underwent a 40-min global ischemia followed by a 60-min reperfusion; IPo group: after a 40-min global ischemia, the process of 10-second reperfusion followed by a 10-second ischemia was repeated 6 times, then there was a continuous 58min reperfusion; 5-hydroxydecanoic acid(5-HD) group: after a 40min global ischemia, hearts with 5HD(100 μmol/L) K-HB were reperfused for 15min and then perfused without 5HD for 45min；IPo+5-HD group: after a 40-min global ischemia, the process that the isolated hearts with 5-HD(100 μmol/L) KHB were reperfused for 10second followed by a 10second ischemia was repeated 6 times, then the hearts with 5-HD(100 μmol/L) KHB were continuously [CM(159mm]perfused for 13-min followed by reperfusion without 5-HD(100 μmol/L) K-HB for 45-min. The cardiac function,coronary flow(CF), cardiac troponin I(cTnI) content in coronary effluent, the area of acute myocardial infarction (AMI) and myocardial ultrastructure were observed. Results Left ventricular developed pressure(74.3±3.3 mm Hg vs. 57.1±3.3 mm Hg,t=1300, P=0.000),+dp/dtmax(1 706.6±135.6 mm Hg/s vs. 1 313.3±96.2 mm Hg/s,t=6.28,P=0.000),－dp/dtmax(1 132.8±112.1 mm Hg/s vs. 575.7±67.7 mm Hg/s,t=13.48, P=0.000) and CF(6.49±0.30 ml/min vs. 3.70±0.24 ml/min,t=28.6,P=0.000) in IPo group were higher than those in I/R group. Left ventricular enddiastolic pressure(10.9±1.7mm Hg vs. 26.2±1.5 mm Hg,t=－19.21, P=0000)and cTnI content in coronary effluent (0.62±0.01 ng/ml vs. 0.71±0.01 ng/ml, t=-12.00,P=0.000) were lower than those in I/R group; the area of AMI decreased 20.8％ compared with that in I/R group (Plt;0.05). The myocardial protective effect in IPo+5HD group was similar with that in IPo group, but lower than that in IPo group. The electron microscope showed that IPo and IPo+5HD could reduce myocardial fiber damage and mitochondrial damage caused by I/R. Conclusion IPo can protect I/R myocardium, which is achieved mainly by activating mitoK-ATP channels.
【摘要】 目的 分析腺苷蛋氨酸治疗慢性乙型肝炎高胆红素血症的临床疗效。 方法 回顾性分析2010年1-12月28例接受腺苷蛋氨酸（2 000 mg静脉滴注，1次/d）治疗慢性乙型肝炎高胆红素血症患者的临床资料，并对腺苷蛋氨酸治疗慢性重症乙型肝炎高胆红素血症后症状、体征及实验室检测指标的改变情况进行总结，利用多因素logistic回归分析方法探索与疗效相关的预测因素。根据相关症状、体征和实验室结果的不同，将疗效分为显效、有效和无效3类。 结果 28例患者使用腺苷蛋氨酸治疗4周后，显效20例（71.4%），有效4例（14.3%），无效4例（14.3%）。多因素logistic回归分析提示病程短、并发症少是影响腺苷蛋氨酸疗效的独立预测因素。 结论 腺苷蛋氨酸是治疗慢性乙型肝炎高胆红素血症有效，发病时间短及并发症少的患者退黄效果更好。【Abstract】 Objective To investigate the curative efficacy of ademetionine in the treatment of hyperbilirubinemia for chronic hepatitis B patients. Methods The clinical data of 28 chronic hepatitis B patients with intrahepatic cholestasis receiving intravenous ademetionine treatment (2 000 mg per day) were retrospectively analyzed. Patients’ symptoms, body signs and laboratory examination results were summarized, and predictors for efficacy were investigated using multiple regression analysis. In this study, the curative efficacy was classified into remarkable efficacy, efficacy and inefficacy, according to the clinical data. Results After one-month treatment with ademetionine, the percentage for remarkable efficacy, efficacy and inefficacy was 71.4%, 14.3%, and 14.3% respectively. Multivariate logistic regression analysis showed that short disease duration and fewer complications were independent predictors for remarkable efficacy of ademetionine treatment. Conclusion Ademetionine is an effective agent for the treatment of hyperbilirubinemia in chronic hepatitis B patients, and the result is especially good for patients with short duration and fewer complications.
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)
Abstract: Objective To observe the combined protective effects of U50 488H and hypothermia preservation on isolated rabbit hearts preconditioned. Methods Forty rabbits were randomly divided into five groups, 8 rabbits in each group. The perfusion model of isolated rabbit hearts was established by the Langendorff device. In the control group: the isolated rabbit hearts were preserved with the University of Wissconsin solution (UW ) for six hours; groupI : the isolated rabbit hearts were preconditioned with St. ThomasII cardioplegic solution containing U50 488H (1. 6mmo l/L ) and then preserved with hypothermic preservation for four hours; groupII ; the precondition was the same as group II , hypothermic preservat ion fo r six hours; group III : the precondit ion was the same as group I , hypothermic preservation for eight hours; group IV : the precondit on was the same as group I , hypothermic preservation for ten hours. The cardiac function, myocardial sarcoplasmic reticulum calcium ion adenosine triphosphatase (SRCa2+ -ATPase) act ivity and calcium ion concentrations in mitochondria were determined at thirty minutes after reperfusion. Results As the hypothermic preservation time increased from four to ten hours, the recovery rate of each index of cardiac function, coronary artery flow (Cf) and SRCa2+ -ATPase activity also decreased, but the calcium ion concentrations in the mitochondria increased. Cardiac function index recovery rates in group I and group II w ere higher than those in group III and groupIV respectively (P lt; 0. 05, 0. 01) ,meanwhile recovery rates of cardiac function index in group III were higher than that in group IV (P lt; 0. 05). Recovery rate of Cf in groupII ( 84. 56%±10. 38%)were higher than those in group III (79. 45%±9. 67% ) and group IV (68. 31%±6. 84% , P lt;0.01) , meanwhile the recovery rate of Cf in group III was higher than that in group IV (P lt; 0. 05). SRCa2+ -A Tpase activity in group II (4. 43±0. 41μmo l/m g?h)were higher than those in control group (3. 04±0. 22Lmo l/mg?h ) , group III (3. 26±0. 29Lmo l/m g?h) and group IV (2. 57±0. 63Lmo l/m g?h, P lt; 0. 05) , SRCa2+ -ATPase activity in group III was higher than that in group IV (P lt; 0. 01). The calcium ion concentrations in mitochondria in group II (38176±4. 30μmo l/g ?dw ) and in the control group (40. 23±3. 75μmol/g ?dw )were less than those in group III (43125±5116μmol/g?dw ) and groupIV (45. 78±3. 26μmol/g?dw , P lt; 0. 05, 0. 01) respect ively. Conclusion The hypothermic preservation time for isolated dono r’s hearts p re-treated with St. Thomas II cardioplegic solution containing U 50 488H should the kep tunder 8h. The myocardial protection effects of both UW solution and U50 488H- containing St. Thomas II cardioplegic solution on isolated dono r’s hearts appear to be the same at 6 hours.
【Abstract】ObjectiveTo investigate the effects of endogenous photodynamic therapy (PDT) on intracellular cAMP and cGMP concentrations of human colon carcinoma cell lines SW480. MethodsSW480 cells were divided into control group, light group, δaminolevulinic acid （ALA） group (ALA group) and endogenous PDT group (ALAPDT group). Intracellular cAMP and cGMP concentrations of each group were detected by radioimmunoassay at 30, 60, 90 and 120 min after irradiation. ResultsThere was a significant increase in intracellular cAMP concentration of ALAPDT group at 30 min after irradiation (P＜0.001) and sequent decrease, but intracellular cAMP concentrations of ALAPDT group at 60, 90 and 120 min after irradiation had no statistical difference than the other groups (Pgt;0.05). Intracellular cGMP concentration of different time point of each group was not significantly different. ConclusionThese results indicate that the cytoprotection of SW480 cell are produced by an instantaneous increase in the intracellular cAMP concentration while endogenous PDT is killing SW480 cell.