ObjectiveTo study the changes of levels of α subunits of stimulatory (Gsα) and inhibitory guanine nucleotide binding protein (Giα) in newborn guinea pig (0 2 days old) myocardium undergoing global ischemic reperfusion, and influences on the changes by St.Thomas Ⅱ and cold blood cardioplegic solution.MethodsThirty newborn guinea pigs were randomly assigned to three groups. GroupⅠ ( n = 10): the newborn hearts suffered by hypothermic global ischemia; group Ⅱ( n =10): the newborn hearts arrested by St. Thomas Ⅱ , and group Ⅲ ( n = 10): the newborn hearts arrested by cold blood cardioplegic solution. Levels of Gsα and Giα were investigated with Western blot analysis.ResultsNo differences of levels of Gsα and Giα were found in three groups before ischemia ( P gt;0.05). The level of Gsα after ischemia was significantly decreased than before ischemia in groupⅠand group Ⅱ ( P lt; 0 01), whereas no pronounced changes in group Ⅲ ( P gt;0.05) were noted after ischemia. The level of Gsα in group Ⅲ was not significantly changed after reperfusion compared with before ischemia( P gt;0 05), and it was much higher than those in groupⅠand group Ⅱ ( P lt; 0 01). Level of Giα was found not markedly changed in group Ⅲ after reperfusion compared with that before ischemia, but was notable higher in groupⅠand group Ⅱ( P lt;0.01). ConclusionsSignificant decrease of level of Gsα, whereas marked increase of level of Giα are found in myocardium of newborn guinea pig undergoing hypothermic (20℃) ischemic reperfusion. No impact of St. Thomas Ⅱ on these changes is verified, but recovery to the level of Gsα and Giα before ischemia is achieved by cold blood cardioplegic solution after ischemia and reperfusion. Unbalance between Gsα and Giα is the one of the mechanisms of ischemic reperfusion injury for immature myocardium.
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 establish an rat model of the Anterior Isc hemic Optic Neuropathy (rAION), and identify its reliability by observing the fundus, fluorescein fundus angiography (FFA),optical coherence tomography (OCT), v isually evoked potential (VEP) and histopathology. Methods Thirty male Sprague-Dawley rats were randomly divided into group Naive with 5 rats, group Laser with 5 rats, group hematoporphyrin derivative(HPD) with 5 rats, group rAION with 15 rats. All of the right eyes were the experimental eyes and the left ones were the control. after administration of HPD in rats` vena caudalis. The rats in group Laser were treated with a krypton red 647nm/2/3disc spot laser for 120 seconds, the rats in group HPD were treated by administration of HPD in rats` vena caudalis, and the rats in group Na?ve were not treated. Results From 1 day to 6 day s after rAION induction, the ON was pale and swollen in the superior part. The ON at 90 days after induction was pale and shrunken.30 minutes after rAION induction, hyperfluoresc ence appeared in the superior part of the optic disc, and the hypofluorescence in the 23rd day. In early FFA, hypofluorescence appeared at the ischemic area of the optic disc, and in midst and later stage the ischemic area revealed hyperflu orescence in the 1st day after rAION induction, the hypofluorescence in midst and later stage in the sixth day after r-AION model. The latent period of F-VEP expanded. The amplitude cut down in the 1-2 days after r-AION induction and did not changed in 35nd day. The surface of optic disc showed higher and rougher tha n the surface of retina in the 6th day after r-AION induction in OCT. After fixation and hematoxylineosin staining of 6-mu;m sections, in high power field the o pt ic disc showed edema with the displacement of retina surrounding the disc 1 day after treatment. Rarefaction and degeneration in the nerve fiber of retina and r eduction of the number of nuclei of ganglion cells in the 23st day after the mod el induction, and the thinning of nerve fiber of the optic disc and its surround ings. In contrast, there was no change in group Na?ve, group Laser and group HPD. Conclusions The r-AION model is like the human AION in fundus, FFA, OCT, VEP and histopathology. The rAION model provides the ischemic changes of occurrence of AION, and is helpful for the fundamental study of the AION. (Chin J Ocul Fundus Dis,2008,24:90-94)
【Abstract】Objective To observe the synthesis of TLR2 protein and its mRNA expression in Kupffer cells (KCs) and sinusoidal endothelial cells(SECs).Methods Thirty-two BALB/c mice divided into two groups (operation group and false operation group) were used to prepare the model of partial hepatic ischemia/reperfusion (I/R) injury. After injury KCs and SECs were isolated with twosteps situ perfusion technique. And these cells were dyed by rat anti-mouse TLR2 IgG and anti-rat IgG2b labeled with flurescein isothiocyanate (FITC). The sysnthesis of TLR2 protein were determined by flow cytometric (FCM) analysis and real time reverse transcription polymerase chain reaction (Real-Time RT-PCR) analysis for gene expression.Results As for KCs: TLR2 expression was significant higher in operation group, compared with false operation group 〔protein expression: （9.19±1.07）% vs （1.52±0.21）%, P＜0.01; gene expression: 0.54±0.77 vs 2.62±2.19, P＜0.05〕. But there were no significant differences with expression in SECs. Conclusion Synthesis of TLR2 protein and its gene expression increased in KCs in the mouse partial hepatic ischemia-reperfusion injury.
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.
Objective To investigate the significance of sensory neuropeptides [calcitonin gene related peptide (CGRP) and substance P (SP)] in steroid-induced avascular necrosis of the femoral head (ANFH) by using a rabbit model. Methods Fifty-five adult female Japanese White rabbits (weighing 3 kg and aging 24 months) were randomly divided into experimental group (n=45) and control group (n=10). The rabbits in experimental group received a single intramuscularinjection of methylprednisolone at a dose of 4 mg/kg and then were sacrificed after 3 days (n=15), 1 week (n=15), and 2 weeks (n=15) of injection. The rabbits in control group were fed without any treatment. The necrosis of the femoral head was observed. And the expressions of the monoclonal antibodies CGRP and SP were observed with immunohistochemical staining. Also, the integrated absorbance (IA) value of the positive area was calculated. Results All rabbits survived to the end of the experiment. There was no necrosis of the bone or bone marrow in experimental group at 3 days; whereas ANFH was observed in 5 rabbits at 1 week (33%) and in 8 rabbits at 2 weeks (53%). There were significant differences in the rate of ANFH between 1 week, 2 weeks and 3 days (P lt; 0.05); but there was no significant difference between 1 week and 2 weeks (P gt; 0.05). The intensity of CGRP immunoreactivity increased and reached the peak at 1 week, and then decreased at 2 weeks in experimental group. The IA value of CGRP in experimental group at 1 week was significantly higher than that of control group and that of experimental group at 3 days (P lt; 0.05). The IA value of CGRP in experimental group at 2 weeks was significantly lower than those at 3 days and 1 week (P lt; 0.05). The intensity of SP immunoreactivity decreased and reached the lowest at 1 week, and then increased. The IA value of SP in experimental group at 1 week was significantly lower than that of control group and that of experimental group at 2 weeks (P lt; 0.05). Conclusion The sensory neuropeptides may be affected by the steroid, which may play a key role in the process of steroid-induced ANFH by imbalance of bone metabol ism, disturbance of the microcirculation of bone, and disorder of the protective pain-transmission.
Objective To investigate the therapeutic effects of the vacularized iliac graft for ischemic necrosis of the femoral head in Niger young patients with sickle cell disease. Methods From November 1998 to Apirl 2001, 12 patients (5 males and 7 females, aging 11-22 years) with sickle cell disease suffered from ischemic necrosis of the femora! head in 14 hips. The lesion was on one hip in 10 patients and on bilateral hip in 2 patients. Necrosis was classified as Ficat Stage Ⅲ-Ⅳ in all patients. Twelve hips in 12...
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.