缺血后处理对高脂血症大鼠局灶性脑缺血再灌注后STAT-3、IL-6表达的影响
发布时间：2018-07-13 14:27
【摘要】：目的:对高脂血症大鼠施行局灶性脑缺血再灌注术,观察缺血后处理对缺血侧脑组织中STAT-3、IL-6及凋亡细胞的动态表达,探讨缺血后处理对高脂血症大鼠脑缺血再灌注损伤的保护作用及可能机制。方法:将115只SD大鼠随机分为两组,其中一组用普通饲料喂养(10只),另一组用高脂饲料喂养(105只),于28天后,空腹心脏采血检测两组大鼠血清中的血脂指标,分别为血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C),高密度脂蛋白(HDL-C),两组含量比较,若存在明显差异(P0.05),表明高血脂血症大鼠造模成功。随机将造模成功的高脂血症大鼠分为假手术组(sham组)35只、缺血再灌注组(I/R组)35只、缺血后处理组(IP组)35只。I/R组与IP组动物模型的制作均使用改良后的线栓法,并在大鼠脑缺血2小时,进行再灌注,术后按时间点的不同将30只大鼠平均分为6组,各时间点的设置分别是3h、6h、12h、24h、48h、72h,IP组于再灌注前给予灌注15s/缺血15s,反复3个循环,sham组仅分离颈动脉后缝合切口。sham组于手术后24小时使用5只大鼠取脑组织行TTC染色,IP组与I/R组于再灌注后24小时使用5只大鼠行TTC染色,计算脑梗死体积。各时间点神经功能缺损情况通过神经功能缺损评分判断,不同时间点脑梗死组织的病理改变通过HE染色观察,神经细胞凋亡的数量采用Tunel法检测,STAT-3和IL-6的表达用免疫组化(Envision法)检测。结果:1.高脂血症大鼠模型:高脂饲料喂养组大鼠血清中TC、TG、LDL-C、HDL-C的含量明显高于普通饲料喂养组(P0.05);2.神经功能缺损评分:sham组大鼠没有出现明显的神经功能缺损症状,I/R组于再灌注不同时间点出现不同程度神经功能障碍,以24小时评分最低,与其他时间点比较差异有统计学意义,IP组在12小时以后神经功能评分显著高于相同时间点I/R组(P0.05);3.TTC染色计算脑梗死体积:sham组术后24小时,TTC染色脑组织为均一的红色,脑组织形态正常,无明显苍白和水肿;I/R组和IP组术后24小时的脑组织内可观察到线栓侧脑组织颜色苍白,体积大于无线栓侧的梗死脑组织,I/R组梗死体积较IP组大(P0.05);4.神经细胞凋亡数量及IL-6、STAT-3免疫阳性细胞的表达:sham组的脑组织经过免疫组化染色后在镜下未见STAT-3免疫阳性细胞,偶可见少量IL-6免疫阳性细胞表达;I/R组脑缺血再灌注后3h,STAT-3开始少量表达,IL-6表达增多,24h达到高峰后开始下降,在72h仍有高表达;IP组STAT-3和IL-6的表达与I/R组变化趋势相似,但数量显著减少(P0.05),凋亡细胞变化与STAT-3、IL-6基本一致。结论:1.在建立高脂血症大鼠模型基础上制作一侧大脑中动脉脑缺血再灌注模型,能更好的模拟人体脑梗死发病模式,使所检测出来的各项指标更加具有说服力;2.脑缺血再灌注后可使大鼠出现行为学及形态学方面的损伤,使IL-6、凋亡细胞及STAT-3表达增多;3.缺血后处理可能的脑保护机制是通过抑制IL-6的表达,进而减少JAK2/STAT3通路的激活,减轻凋亡程度。
[Abstract]:Objective: To observe the dynamic expression of STAT-3, IL-6 and apoptotic cells in ischemic brain tissue after ischemic reperfusion in rats with hyperlipidemia, and to explore the protective effect and possible mechanism of ischemic postconditioning on cerebral ischemia reperfusion injury in hyperlipidemia rats. Methods: 115 rats were randomly divided into two groups, of which 115 rats were randomly divided into groups. One group was fed with ordinary feed (10), the other was fed with high fat diet (105). After 28 days, blood serum lipid indexes in two groups of rats were detected by empty heart blood sampling, which were serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C), and there were significant differences (P0.05) in the two groups (P0.05). The rats with hyperlipidemia were successfully built. The rats were randomly divided into 35 rats in the sham operation group (Group sham) and 35 rats in the ischemia reperfusion group (group I/R). The 35 group of.I/R and IP animal models in the ischemic postconditioning group (group IP) all used the modified thread thrombus method and reperfusion for 2 hours of cerebral ischemia in rats, and on time after the operation. The difference between the 30 rats was divided into 6 groups, and the setting of each time point was 3h, 6h, 12h, 24h, 48h, 72h. Before reperfusion, group IP was given 15s/ ischemia 15s and repeated 3 cycles. The sham group only separated the carotid artery and the.Sham group took 5 rats for TTC staining at 24 hours after the operation. At 24 hours, 5 rats were stained with TTC to calculate the volume of cerebral infarction. The nerve function defect in each time point was judged by the nerve function defect score. The pathological changes of cerebral infarction in different time points were observed by HE staining. The number of neuronal apoptosis was detected by Tunel, and the expression of STAT-3 and IL-6 was detected by immunohistochemical (Envision) method. Results: 1. the model of hyperlipidemia rats: the content of TC, TG, LDL-C, HDL-C in the serum of the high fat feed group was significantly higher than that of the normal feed group (P0.05). 2. the score of nerve function defect: the rats in the group sham did not have the obvious symptoms of nerve function defect, and the I/R group appeared different degree of nerve dysfunction at different time points of reperfusion in the I/R group. The score of 24 hours was lowest, and the difference was statistically significant compared with other time points. After 12 hours, the score of nerve function in group IP was significantly higher than that in group I/R (P0.05); 3.TTC staining was used to calculate cerebral infarction volume: 24 hours after operation in group sham, TTC staining brain tissue was uniform red, brain tissue was normal, no obvious paleness and edema; I/R group; I/R group. In group IP, the brain tissue of 24 hours after operation could be observed that the color of the brain tissue was pale and the volume was larger than that of the infarct brain tissue on the side of the radio. The infarct volume in group I/R was larger than that of the IP group (P0.05); the number of apoptosis and the expression of IL-6, STAT-3 immunoreactive cells were expressed in the group of sham: the brain tissue of group sham had no STAT-3 exemption under the microscope after immunohistochemical staining. The expression of a small amount of IL-6 immunoreactive cells was found in the pestilence positive cells. In group I/R, 3h, STAT-3 began to express a small amount of expression, IL-6 expression increased, 24h began to decrease after the peak, and still had high expression in 72h; the expression of STAT-3 and IL-6 in IP group was similar to that in I/R group, but the number of apoptotic cells decreased significantly (P0.05). Conclusion: 1.: 1. on the basis of the establishment of hyperlipidemia rat model, the model of cerebral ischemia reperfusion in one side of the cerebral artery can better simulate the pattern of cerebral infarction and make the detected indexes more convincing. 2. after cerebral ischemia reperfusion, it can cause the injury of behavior and morphology in rats. Increase the expression of IL-6, apoptotic cells and STAT-3; 3. the possible mechanism of cerebral protection after ischemic post-treatment is to reduce the expression of IL-6, then reduce the activation of JAK2/STAT3 pathway and reduce the degree of apoptosis.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3
本文编号：2119690
[Abstract]:Objective: To observe the dynamic expression of STAT-3, IL-6 and apoptotic cells in ischemic brain tissue after ischemic reperfusion in rats with hyperlipidemia, and to explore the protective effect and possible mechanism of ischemic postconditioning on cerebral ischemia reperfusion injury in hyperlipidemia rats. Methods: 115 rats were randomly divided into two groups, of which 115 rats were randomly divided into groups. One group was fed with ordinary feed (10), the other was fed with high fat diet (105). After 28 days, blood serum lipid indexes in two groups of rats were detected by empty heart blood sampling, which were serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C), and there were significant differences (P0.05) in the two groups (P0.05). The rats with hyperlipidemia were successfully built. The rats were randomly divided into 35 rats in the sham operation group (Group sham) and 35 rats in the ischemia reperfusion group (group I/R). The 35 group of.I/R and IP animal models in the ischemic postconditioning group (group IP) all used the modified thread thrombus method and reperfusion for 2 hours of cerebral ischemia in rats, and on time after the operation. The difference between the 30 rats was divided into 6 groups, and the setting of each time point was 3h, 6h, 12h, 24h, 48h, 72h. Before reperfusion, group IP was given 15s/ ischemia 15s and repeated 3 cycles. The sham group only separated the carotid artery and the.Sham group took 5 rats for TTC staining at 24 hours after the operation. At 24 hours, 5 rats were stained with TTC to calculate the volume of cerebral infarction. The nerve function defect in each time point was judged by the nerve function defect score. The pathological changes of cerebral infarction in different time points were observed by HE staining. The number of neuronal apoptosis was detected by Tunel, and the expression of STAT-3 and IL-6 was detected by immunohistochemical (Envision) method. Results: 1. the model of hyperlipidemia rats: the content of TC, TG, LDL-C, HDL-C in the serum of the high fat feed group was significantly higher than that of the normal feed group (P0.05). 2. the score of nerve function defect: the rats in the group sham did not have the obvious symptoms of nerve function defect, and the I/R group appeared different degree of nerve dysfunction at different time points of reperfusion in the I/R group. The score of 24 hours was lowest, and the difference was statistically significant compared with other time points. After 12 hours, the score of nerve function in group IP was significantly higher than that in group I/R (P0.05); 3.TTC staining was used to calculate cerebral infarction volume: 24 hours after operation in group sham, TTC staining brain tissue was uniform red, brain tissue was normal, no obvious paleness and edema; I/R group; I/R group. In group IP, the brain tissue of 24 hours after operation could be observed that the color of the brain tissue was pale and the volume was larger than that of the infarct brain tissue on the side of the radio. The infarct volume in group I/R was larger than that of the IP group (P0.05); the number of apoptosis and the expression of IL-6, STAT-3 immunoreactive cells were expressed in the group of sham: the brain tissue of group sham had no STAT-3 exemption under the microscope after immunohistochemical staining. The expression of a small amount of IL-6 immunoreactive cells was found in the pestilence positive cells. In group I/R, 3h, STAT-3 began to express a small amount of expression, IL-6 expression increased, 24h began to decrease after the peak, and still had high expression in 72h; the expression of STAT-3 and IL-6 in IP group was similar to that in I/R group, but the number of apoptotic cells decreased significantly (P0.05). Conclusion: 1.: 1. on the basis of the establishment of hyperlipidemia rat model, the model of cerebral ischemia reperfusion in one side of the cerebral artery can better simulate the pattern of cerebral infarction and make the detected indexes more convincing. 2. after cerebral ischemia reperfusion, it can cause the injury of behavior and morphology in rats. Increase the expression of IL-6, apoptotic cells and STAT-3; 3. the possible mechanism of cerebral protection after ischemic post-treatment is to reduce the expression of IL-6, then reduce the activation of JAK2/STAT3 pathway and reduce the degree of apoptosis.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.3
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