Objective: To explore the expression of hypoxia inducible factor-1α (HIF-1α) and the correlation between HIF-1α and apoptosis after traumatic brain injury. Methods: Using experimental traumatic brain injury in the ...Objective: To explore the expression of hypoxia inducible factor-1α (HIF-1α) and the correlation between HIF-1α and apoptosis after traumatic brain injury. Methods: Using experimental traumatic brain injury in the rats, the expression of HIF-1α was studied by immunohisto-chemistry in cerebral tissue, apoptotic cell death was evaluated with TUNEL (transferase-mediated X-dUTP nick end labeling), and double-labeled immunohistochemistry and TUNEL methods were used to investigate the relationship between HIF-1α and apoptosis. Results: There was remarkable difference in the expression of HIF-1α between the experimental groups and the control groups (P< 0.01), in the experimental groups, the expression of HIF-1α at 48 hours was highest; the evidence of apoptotic cell death after experimental traumatic brain injury was found by TUNEL; the apoptotic percentage increased or decreased according to the changes of the positive expression of HIF-1α (r= 0.99). Conclusions: The results suggest that secondary brain ischemia plays a crucial role in apoptotic cell death after traumatic brain injury; HIF-1α can prompt apoptotic cell death after experimental traumatic brain injury.展开更多
To explore the variant processes of cell apoptosis and the inhibiting effect of moderate hypothermia on cell apoptosis after diffuse brain injury. Methods: Models of diffuse brain injury were induced by the trauma dev...To explore the variant processes of cell apoptosis and the inhibiting effect of moderate hypothermia on cell apoptosis after diffuse brain injury. Methods: Models of diffuse brain injury were induced by the trauma device reported by Marmarou. 1 A total of 128 Wistar rats were divided into 4 groups: the uninjured group (Group A, n=8), the severely injured group (Group B, n=60), the mildly injured group (Group C, n=30) and the mild hypothermia group (Group D, n=30). In Group D, the severely injured rats were treated with moderate hypothermia to keep the rectal temperature at 32℃ (standard deviation for 0.1 ℃) for 6 hours. Then the morphosis, the characteristics and the quantity of apoptotic cells in the cerebral cortex and in the hippocampus regions after different severities of craniocerebral injuries were observed and compared under an electronic microscope, with terminal deoxynucleotidyl nick end labeling (TUNEL) in DNA fragmentation and with agarose gel electrophoresis. Results: TUNEL showed apoptotic cells increased according to the injury severity, and they peaked at 48 hours after injury and then declined. In Group C, apoptosis was located in the CA 2 and CA 3 areas of the hippocampus. And in Group B, apoptosis increased evidently, and located in the whole hippocampus and in the frontal and parietal cortex regions. The hypothermia treated rats had some apoptotic cells, too. However, even at 24, 48 and 72 hours after injury there were significantly fewer apoptotic cells in the cortex and in the hippocampus in Group D than that in the non treated groups. Electron microscopy showed that the apoptotic cells were round and shrunken in morphology and the nuclei were round and condensed at 24 and 48 hours after injury. And the apoptosis at 48 hours was more severe than that at 24 hours. The hypothermia treated rats had no apoptotic cells. Gel electrophoresis showed that characteristic DNA “ladders” were observed in the cortex and in the hippocampus at 48 hours after severe injury. But there was no DNA “ladder” at other time points in the severely injured group, in the mildly injured group and in the hypothermia treated group. Conclusions: It suggests that apoptosis occurs after diffuse brain injury and apoptotic cells increase with the injury severity. Moderate hypothermia has a specific inhibiting effect on cell apoptosis after diffuse brain injury in rats.展开更多
Objective： To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury（CIRI）...Objective： To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury（CIRI） rats. Methods： Sixty healthy male Sprague-Dawley（SD） rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion（MCAO） model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride（TTC） staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.Results： Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences（all P〈0.05）. After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3（all P〈0.05）, and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group. Conclusion： Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.展开更多
Objective： To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury （CIRI）, and the influence on protein expression levels of phosphor...Objective： To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury （CIRI）, and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK3/2 in the mitogen-activated protein kinase （MAPK）/extracellular regulated protein kinases （ERK） pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke. Methods： Ninety Sprague-Dawley （SD） rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion （MCAO） models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride （TTC） staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK3/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay. Results： Compared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK：1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant （P〈0.05 or P〈0.01）. Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphowlated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant （P〈0.05 or P〈0.01）. Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK3/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant （P〈0.05 or P〈0.01）. Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant （P〈0.01）. Conclusion： Acupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-l, MEK-2 and ERK：1/2 protein expression levels.展开更多
基金JiangsuProvinceTechnology AssociationFoundation (No .DJ995 0 1)
文摘Objective: To explore the expression of hypoxia inducible factor-1α (HIF-1α) and the correlation between HIF-1α and apoptosis after traumatic brain injury. Methods: Using experimental traumatic brain injury in the rats, the expression of HIF-1α was studied by immunohisto-chemistry in cerebral tissue, apoptotic cell death was evaluated with TUNEL (transferase-mediated X-dUTP nick end labeling), and double-labeled immunohistochemistry and TUNEL methods were used to investigate the relationship between HIF-1α and apoptosis. Results: There was remarkable difference in the expression of HIF-1α between the experimental groups and the control groups (P< 0.01), in the experimental groups, the expression of HIF-1α at 48 hours was highest; the evidence of apoptotic cell death after experimental traumatic brain injury was found by TUNEL; the apoptotic percentage increased or decreased according to the changes of the positive expression of HIF-1α (r= 0.99). Conclusions: The results suggest that secondary brain ischemia plays a crucial role in apoptotic cell death after traumatic brain injury; HIF-1α can prompt apoptotic cell death after experimental traumatic brain injury.
文摘To explore the variant processes of cell apoptosis and the inhibiting effect of moderate hypothermia on cell apoptosis after diffuse brain injury. Methods: Models of diffuse brain injury were induced by the trauma device reported by Marmarou. 1 A total of 128 Wistar rats were divided into 4 groups: the uninjured group (Group A, n=8), the severely injured group (Group B, n=60), the mildly injured group (Group C, n=30) and the mild hypothermia group (Group D, n=30). In Group D, the severely injured rats were treated with moderate hypothermia to keep the rectal temperature at 32℃ (standard deviation for 0.1 ℃) for 6 hours. Then the morphosis, the characteristics and the quantity of apoptotic cells in the cerebral cortex and in the hippocampus regions after different severities of craniocerebral injuries were observed and compared under an electronic microscope, with terminal deoxynucleotidyl nick end labeling (TUNEL) in DNA fragmentation and with agarose gel electrophoresis. Results: TUNEL showed apoptotic cells increased according to the injury severity, and they peaked at 48 hours after injury and then declined. In Group C, apoptosis was located in the CA 2 and CA 3 areas of the hippocampus. And in Group B, apoptosis increased evidently, and located in the whole hippocampus and in the frontal and parietal cortex regions. The hypothermia treated rats had some apoptotic cells, too. However, even at 24, 48 and 72 hours after injury there were significantly fewer apoptotic cells in the cortex and in the hippocampus in Group D than that in the non treated groups. Electron microscopy showed that the apoptotic cells were round and shrunken in morphology and the nuclei were round and condensed at 24 and 48 hours after injury. And the apoptosis at 48 hours was more severe than that at 24 hours. The hypothermia treated rats had no apoptotic cells. Gel electrophoresis showed that characteristic DNA “ladders” were observed in the cortex and in the hippocampus at 48 hours after severe injury. But there was no DNA “ladder” at other time points in the severely injured group, in the mildly injured group and in the hypothermia treated group. Conclusions: It suggests that apoptosis occurs after diffuse brain injury and apoptotic cells increase with the injury severity. Moderate hypothermia has a specific inhibiting effect on cell apoptosis after diffuse brain injury in rats.
基金supported by National Natural Science Foundation of China,No.81303051Traditional Chinese Medicine Science Research Planning Project of Hunan Province,No.201471Province and Ministry Co-construction Key Laboratory for Internal Medicine of Traditional Chinese Medicine of the Education Ministry of China,No.ZYNK201501~~
文摘Objective： To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury（CIRI） rats. Methods： Sixty healthy male Sprague-Dawley（SD） rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion（MCAO） model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride（TTC） staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.Results： Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences（all P〈0.05）. After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3（all P〈0.05）, and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group. Conclusion： Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.
基金supported by National Natural Science Foundation of China(No.81303051)Traditional Chinese Medicine Science Research Planning Project of Hunan Province(No.201471)Province and Ministry Co-construction Key Laboratory for Internal Medicine of Traditional Chinese Medicine of the Education Ministry of China(No.ZYNK201501)~~
文摘Objective： To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury （CIRI）, and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK3/2 in the mitogen-activated protein kinase （MAPK）/extracellular regulated protein kinases （ERK） pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke. Methods： Ninety Sprague-Dawley （SD） rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion （MCAO） models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride （TTC） staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK3/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay. Results： Compared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK：1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant （P〈0.05 or P〈0.01）. Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphowlated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant （P〈0.05 or P〈0.01）. Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK3/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant （P〈0.05 or P〈0.01）. Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant （P〈0.01）. Conclusion： Acupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-l, MEK-2 and ERK：1/2 protein expression levels.
