目的研究依達拉奉影響肝臟缺血再灌注過程中TNF-α的表達情況，探討依達拉奉對肝臟缺血再灌注損傷的逆轉作用。 方法將80只Wistar大鼠編號，根據計算機產生隨機數字，前40為一組，后40為一組，分為實驗組和對照組2組，建立常溫下部分肝缺血再灌注損傷動物模型。 在肝臟缺血再灌注損傷開始前1 h和開始時對實驗組大鼠給予依達拉奉注射液10 ml，對照組則給予同等容量的生理鹽水。分別于再灌注后0、1、2及4 h測定肝臟脂質過氧化物酶（LPO）和肝臟谷草轉氨酶(AST) 濃度； 應用RT-PCR法檢測肝組織TNF-α mRNA含量，并測定肝組織和血清中TNF-α水平； 應用TUNEL染色法檢測缺血肝組織的細胞凋亡情況。結果再灌注后1、2及4 h，實驗組大鼠肝臟LPO及AST濃度均明顯低于對照組（Plt;0.001）； 實驗組再灌注后1 h時肝組織TNF-α mRNA表達量、肝組織和血清TNF-α含量均明顯升高且達峰值，但均明顯低于對照組（Plt;0.05）； 再灌注后各時相實驗組肝細胞凋亡率明顯升高，但均明顯低于對照組（Plt;0.05）。 結論依達拉奉能抑制氧化應激反應，從而降低肝缺血再灌注損傷； 并顯著減少炎性細胞因子TNF-α的產生，抑制炎性反應的發生，減少肝細胞的凋亡。
Objective To study the effect of Kupffer cell on the liver ischemia/reperfusion injury.Methods The literature in recent years on the liver ischemia/reperfusin injury were reviewed.Results The activated kupffer cell can generate and release a variety of soluble toxic mediators, affect the liver microcirculation directly or indirectly. Conclusion Kupffer cell have important effect on liver ischemia/reperfusion injury.
To investigate the protective effect of propofol on ischemia/reperfusion induced spinal cord injury in rabbits and its influence on excitatory amino acid (EAA). Methods Sixty New Zealand white rabbits weighing 2.0-2.5 kg, half males and half females, were selected. The infrarenal circumaortic clamping model was used. And 6 mL/kg different fluids were continuously infused through a catheter into the aorta distal to the clamping site at a speed of 12 mL/(kg?h) during the 30 minutes ischemia period. According to the different infusing l iquids, the rabbits were randomized into 6 groups(n=10 per group): group A, normal sal ine; group B, 10% intral ipid; group C, propofol 30 mg/kg; group D, propofol 40 mg/kg; group E, propofol 50 mg/kg; group F, propofol 60 mg/kg. At 0, 6, 24, and 48 hours after reperfusion, neurologic outcomes were scored on a Tarlov scale system. At 48 hours after reperfusion, the number of normal neurons in the anterior spinal cord was counted, and concentration of EAA in the lumbar spinal cord was measured by high performance l iquid chromatography. Results The neuroethological score was better in groups C, D, E and F than that of groups A and B (P lt; 0.05), the score of group E was the highest (P lt; 0.05), and there was no significant difference between group A and group B (P gt; 0.05). The number of normal neurons in the anterior spinal cord of groups C, D, E and F was greater than that of groups A and B (P lt; 0.05), and group E was greater than groups C, D and F (P lt; 0.05). The concentration of EAA in groups A, B, C, D, E and F was greater than that of normal tissue, the group E was the lowest (P lt; 0.05), the groups A and B were the highest (P lt; 0.05), and there was no significant difference between group A and group B (P gt; 0.05). Concentrations of glutamate and aspartic acid were negatively correlated to normal neuron numbers in the anterior spinal cord and neuroethological scores 48 hours after reperfusion, and the corresponding correlation coefficient was — 0.613, — 0.536, — 0.874 and — 0.813, respectively (P lt; 0.01). Conclusion Propofol can significantly inhibit the accumulation of EAA in spinal cord and provide a protective effect against the ischemia/reperfusion injury induced spinal cord in rabbits.
OBJECTIVE: To investigate the injury on isolated testes induced by ischemia/reperfusion(I/R), and the protective effect of Yisheng injection on the injury. METHODS: Twenty-six isolated cadaver testes contributed by 13 persons were preserved with 4 degrees C 250 ml hypertonic citrate alloxuric (HCA) solution and then reperfused with 37 degrees C 500 ml HCA. Solution of experimental group contained 500 micrograms/ml Yisheng injection. In simple cold preservation test, involving in 8 experimental and 8 control testes, a series of time points (6, 12, 18, 24, 36, 48, 60, 72 hours) were set to harvest. 10 testes (1 testis respectively on 6, 12, 18, 24 and 36 hours in experimental and control groups) were reperfused with 37 degrees C HCA for 6 and 12 hours. Histological and histochemical changes were observed. RESULTS: In the experimental testes, 4 degrees C cold preservation in 24 hours could not induce obvious pathologic changes. After 24 hours, changes such as swelling, vacuolar degeneration or detachment of endothelial cells (ECs), separation between basement membrane and seminiferous epithelium, mal-alignment of spermatogenous cell and edema of mesenchyme could be observed. In the testes preserved for 12 hours, the activity of lactic dehydrogenase(LDH) and succinic dehydrogenase (SDH) increased, then fallen after 24 hours. The activity of Nitric oxide synthetase(NOS) decreased after 18 hours. All changes were more obvious after following 37 degrees C reperfusion. In the control testes, swelling and vacuolar degeneration of ECs occurred on 12 hours cold preservation, and injury was worse along with the prolongation of cold preservation time. Pathologic changes of ECs, seminiferous epithelium and mesenchyme were serious after 37 degrees C reperfusion. CONCLUSION: 4 degrees C cold preservation in 24 hours can only cause mild ECs’ injury, and obvious abnormal testes’ histological profile can be observed beyond 24 hours. 37 degrees C reperfusion will make injury worse. Yisheng injection can keep isolated testes histologic structure well in 24 hours cold preservation, and it has protective effect on I/R injury.
Objective To investigate the extent of hepatic ischemia reperfusion (HIR) injury in rat cirrhotic liver under different ischemic time，and find the time limit under which the rat with cirrhotic liver could tolerate. Methods At first，the cirrhosis of the rat were induced by carbon tetrachloride（CCl4）injected subcutaneously. Then these rats were randomly divided into four groups. Group A(n=6) was made by sham operation, group B, C, D(n=16) were respectively given 20, 30, 40min hepatic warm ischemia. The 7day survival rate, AST, ALT, TNF and liver, pulmonary pathology were observed. Results The 7-day survival rate was decreased with the increase of hepatic ischemic time. The survival rate of group B, C, D were respectively 100%, 60%, 40%. Between group C, D and group B there were significant differences（P＜0.05）. The level of AST and ALT in group D were （2 448.4±942.3）u/L and （1 189.0±403.4）u/L respectively, and those in group C were （2 185.1±1 732.9）u/L and （1 183.5±707.2）u/L respectively, which were higher than those in group B and A significantly（P＜0.01）. The level of TNF was increased significantly 4hr after reperfusion, as compared with that before operation 〔(0.177±0.139）u/ml〕, P＜0.01. TNF of group B, C, D were （0.399±0.216）u/ml, （0.671±0.351）u/ml and （0.789±0.371）u/ml respectively. At the same time the level of TNF in group C, D was higher than that in group B, A significantly（P＜0.01）. Liver and lung pathology showed increased damage with increasing ischemia. Conclusion Hepatic injury is induced by HIR in rats with cirrhotic liver, and its severity increases with the increase of ischemic time. There is a certain hepatic ischemic time between 20min and 30min, which can be tolerated by the rats with cirrhotic liver. TNF may be used as an indicator，showing the degree of HIR injury and foreseeing the result of injury.
【摘要】 目的 通過建立活體大鼠心肌缺血再灌注模型，模擬人類冠心病，研究聚合血紅蛋白（PolyHb）在心肌缺血再灌注中的保護作用，探究PolyHb在冠心病領域中的保護和治療作用。 方法 將45只Sprague-Dawley（SD）大鼠隨機分成3組:實驗組（15只）、對照組（15只）、假手術組（15只），建立大鼠心肌缺血模型。實驗組建立動物模型后，結扎冠狀動脈35 min，并于結扎后5 min，通過SD大鼠尾靜脈按1 mL/min的速度注射1 mL（100 g/L）的PolyHb溶液。缺血完成后開放灌注120 min。對照組建立動物模型，結扎冠狀動脈35 min，并于結扎后5 min，通過SD大鼠尾靜脈按1 mL/min的速度注射1 mL生理鹽水。缺血完成后開放灌注120 min。假手術組僅建立動物模型，但不結扎冠狀動脈，也不再灌注。比較3組SD大鼠的血流動力學參數左室內壓最大上升和下降速率、心肌酶（血清肌酸激酶、乳酸脫氫酶）及病理學檢查（梗死心肌占總心肌面積的百分比），來衡量PolyHb的作用。 結果 與對照組比較，用PolyHb處理的實驗組可增強再灌注時左室內壓最大上升和下降速率（Plt;0.05），減少血液中血清肌酸激酶和乳酸脫氫酶的含量（Plt;0.05），并明顯減少心肌梗死面積百分比（Plt;0.05）。 結論 在心肌缺血的SD大鼠中，PolyHb可以有效的降低缺血再灌注損傷，從而達到心肌保護作用。【Abstract】 Objective To investigate the protective effect of polymerized hemoglobin (PolyHb) for myocardial ischemia-reperfusion and explore the field of polymerized hemoglobin in the protection and treatment of human coronary heart disease, by simulating human coronary heart disease and establishing myocardial ischemia-reperfusion model in living rats. Methods Forty-five Sprague-Dawley (SD) rats were randomly divided into 3 groups: experimental group (n=15), control group (n=15), and sham operation (SHAM) group (n=15). Rat models of myocardial ischemia-reperfusion were established. For the rats in the experimental group, we ligated their left coronary artery for 35 minutes, and injected 1 mL (100 g/L) PolyHb solution into their body at a speed of 1 mL/min 5 minutes later. After ischemia was completed, reperfusion was performed for 120 minutes. The procedures carried out for the rats in the control group were exactly the same except that the PolyHB solution was replaced by 1 mL saline solution. Ligation of the artery and reperfusion were not performed on the rats in the SHAM group. Hemodynamic parameters (maximal rising and falling rates of left ventricular pressure), enzymes (serum creatine kinase, lactate dehydrogenase) and results of histopathologic examinations (percentage of myocardial infarction area over the total myocardial area) were measured and compared among the three groups to evaluate the function of PolyHb. Results Compared with the control group, the experimental group treated with PolyHb had higher maximum rising and falling rates of left ventricular pressure (Plt;0.05), lower blood levels of creatine kinase and lactate dehydrogenase (Plt;0.05), and lower percentage of myocardial infarction area over the total myocardial area (Plt;0.05). Conclusion Polymerized hemoglobin can effectively reduce the ischemia-reperfusion injury and achieve myocardial protection in SD rats with myocardial ischemia.
Objective To study the effect of dexamethasone to protect flaps from an ischemia-reperfusion injury and elucidate its mechanism of regulating the death course of the neutrophils.Methods The rats were randomly divided into 3 groups.The vein of the rat was clamped for 8 h after the flap had formed. Group A: the normal flap; Group B: the saline control flap; Group C: the treatment flap with dexamethasone. The survival area of the flaps was measured at 7 days; the apoptotic and necrotic neutrophils,tumor necrosis factor α （TNF-α）, and interleukin 10 （IL-10） concentrations were measured. Results The flap survival areas in Groups A and C were larger than those in Group B. The apoptotic neutrophils in Group B were fewer than those in Groups A and C on the 1st and 3rd days after operation; however, they were more in number in Group B than in groups A andC on the 6th day. The necrotic cells in Group B were more in number than those in Groups A and C. In Group B, the plasma TNF-α concentration reached the maximum level at 1 h,while the IL-10 level reached the lowest 3 h after the reperfusion. In Group C, the TNF-α concentration was lower than that in Group B and decreased dramatically at 6 h. The IL-10 concentration was the lowest at 1 h, and increased rapidly at 3 h. Thus, ischemia reperfusion could injure the flaps, probably through the abnormal action of the neutrophils, such as the disordered secretion of the cytokines and abnormal death course of the neutrophils. Conclusion Dexamethasone can protect the flap from an ischemia-reperfusion injury by its regulation for the neutrophil function.
Objective To summarize recent research advancement on gene therapy for hepatic ischemia-reperfusion injury (IRI). Methods Relevant references about basic and clinical researches of hepatic IRI were collected and reviewed. Results Recent experimental researches indicated that the expression of several genes and cytokines could protect hepatic cells by suppressing cell apoptosis, decreasing the production of oxyradical, remaining and improving portal venous flow, promoting bilifaction, self immunoloregulation and decreasing inflammatory reaction, so that it could decrease IRI. Conclusion IRI could be decreased by regulating the expressing of target genes or transducing relative genes in vivo, but the path of gene transfer and the selection and optimization of gene carrier still need more basic and clinical researches to prove.
Objective To compare the effect of two types of intermittent pressure on formation of pressure ulcer in rabbit hind l imbs and to investigate the mechanism of gradually changed intermittent pressure produced by waves bed in the prevention of pressure ulcer. Methods Gracil is (3 cm2) in both hind l imbs of 12 adult Japanese white rabbits were randomlyloaded with gradually changed intermittent pressure (50-160 mm Hg, 1 mm Hg=0.133 kPa) and sustained pressure (100 mmHg) serving as the experimental group and the control group, respectively. The experiment was terminated after 4 cycles, and a single cycle included 2 hours of compression and 30 minutes of compression-release. Blood velocity of hind l imbs and blood perfusion of wound were detected by bidirectional doppler blood flow detector and laser doppler perfusion imaging detection system before compression and at every 10 minutes in compression-release period of each cycle (0, 10, 20 and 30 minutes). After the termination, gross observation of the wound was conducted, pathomorphological changes of tissues from compressed area were observed by HE staining, and contents of NO, malondialdehyde (MDA), and superoxide dismutase (SOD) in muscle tissue were measured using colorimetry method. Results No significant difference was evident between two groups in terms of blood flow velocity before compression (P gt; 0.05); the blood flow velocity of two groups decreased significantly at 0 minute in every compressionrelease period of each cycle, and no significant differences were noted between two groups (P gt; 0.05); the blood flow velocity of theexperimental group was higher than that of the control group at 10, 20 and 30 minutes (P lt; 0.05). No significant difference was noted between two groups in terms of wound blood perfusion before compression (P gt; 0.05); the wound blood perfusion of two groups decreased significantly at 0 minute in every compression-release period of each cycle, and no significant differences were noted between two groups (P gt; 0.05); the difference between two groups was not significant at 10 minutes in the first cycle (P gt; 0.05), and the experimental group was higher than the control group at 20 and 30 minutes in the first cycle (P lt; 0.05). In the following 3 cycles, the recovery of perfusion in the experimental group was faster than that of the control group (P lt; 0.05). Gross observation showed the experimental group had less effusion than the control group. The experimental group had intact cutaneous appendage, less inflammatory cell infiltration, and no obvious ulcer formation, whereas the control group had obvious skin ulcer, depletion of cutaneous appendage, and more inflammatory cells infiltration. Significant differences were noted between two groups in terms of NO, MDA, and SOD content (P lt; 0.05). Conclusion Gradually changed intermittent pressure can maintain the blood perfusion of tissue, reduce ischemia-reperfusion injury and cell apoptosis, and prevent the formation of pressure ulcer.
目的 探討丙泊酚-瑞芬太尼對肝臟缺血再灌注損傷的保護作用以及作用機制。 方法 2009年6月－2011年12月選擇擇期需阻斷肝門的肝臟手術患者40例，隨機分為丙泊酚-瑞芬太尼組（P組）和異氟醚組（I組），每組20例。在術前（T0）和肝門阻斷開放后30 min（T1）、60 min（T2）、6 h（T3）、24 h（T4）、72 h（T5）分別抽取動脈血，測定天冬氨酸氨基轉移酶（AST）、丙氨酸氨基轉移酶（ALT）和腫瘤壞死因子α（TNF-α）的含量。 結果 兩組AST、ALT、TNF-α較術前均有增高，差異有統計學意義（P＜0.05）；P組增高幅度明顯低于I組，差異有統計學意義（P＜0.05）。 結論 丙泊酚-瑞芬太尼對肝臟缺血再灌注損傷具有保護作用，抑制TNF-α的產生可能為其作用機制之一。