TLR4-MyD88信号通路与大鼠远程缺血后适应脑保护作用关系的实验研究

发布时间：2018-05-20 15:38

本文选题：远程缺血后适应 + TLR4-MyD88　；参考：《河北联合大学》2014年硕士论文

【摘要】：目的观察大鼠远程缺血后适应时海马CA1区TLR4、MyD88、NF-κB、hs-CRP、IL-6表达变化，探讨TLR4-MyD88信号转导通路在大鼠远程缺血后适应中的脑保护作用。方法172只健康雄性SD大鼠（体重250g~300g）随机分为模型组（n=24）；实验组，即远程缺血后适应组（n=24）；假手术组（n=24）。各组分为12h、48h、24h、72h4个时间点组（n=6）。2应用改进的Longa法，建立大鼠大脑中动脉阻塞（MCAO）的局部脑缺血再灌注模型。实验组于大鼠MCAO2h后再灌注，并即刻给予远程缺血后适应处理。3HE染色观察海马病理学变化；使用免疫组织化学染色检测海马CA1区TLR4、MyD88、NF-κB、hs-CRP、IL-6的表达。4组织切片采用OLYMPUS摄像显微镜（×200）摄像，并应用Motic Med6.0数码医学图像分析系统进行分析。5所得数据均采用SPSS17.0统计软件进行分析，计量资料用均数±标准差（x±s）表示，做单因素方差分析，以P＜0.05为差异有统计学意义。应用Pearson相关系数分析各检测因素相关性，r≠0为具有相关性。结果1HE染色假手术组大鼠海马结构清晰，CA1区神经元形态正常，排列整齐、层次分明；模型组大鼠海马结构松散，CA1区神经元缺失明显，可见较多神经元胞体缩小、核固缩深染，也有胞体肿胀、胞浆内形成空泡的神经元以及小片状坏死区域；实验组神经元缺失、细胞肿胀较模型组明显减轻，细胞形态趋于改善。 2假手术组海马CA1区，各时间点TLR4表达水平均较低。模型组与实验组TLR4表达水平较假手术组明显增高（P＜0.05）。实验组TLR4阳性表达在72h前均低于模型组（P＜0.05），而72h时略高于模型组，但无统计学意义（P＞0.05）。3假手术组海马CA1区，各时间点MyD88表达水平均较低。模型组与实验组MyD88表达明显增高（P＜0.05）。实验组MyD88阳性表达在各时间点均低于模型组（P＜0.05）。4假手术组海马CA1区，各时间点NF-κB表达水平均较低。模型组与实验组NF-κB表达均较假手术组明显增高（P＜0.05）。实验组NF-κB阳性表达在各时间点均略低于模型组，但无统计学意义（P＞0.05）。5假手术组海马CA1区，各时间点hs-CRP仅有少量表达，模型组与实验组hs-CRP表达较假手术组明显增高（P＜0.05），实验组hs-CRP阳性表达在各时间点均明显低于模型组（P＜0.05）。假手术组海马CA1区，各时间点IL-6仅有少量表达，模型组与实验组IL-6表达较假手术组明显增高（P＜0.05），实验组IL-6阳性表达在各时间点均低于模型组（P＜0.05）。6在模型组中TLR4与MyD88表达水平成正相关，但相关性并不显著（r=0.213，P=0.317）；MyD88与NF-κB表达水平成正相关，，但相关性不显著（r=0.302，P=0.151）；NF-κB与IL-6表达呈显著正相关（r=0.792，P＜0.001）；NF-κB与hs-CRP表达也呈显著正相关（r=0.767，P＜0.001）。结论缺血再灌注损伤时大鼠海马CA1区TLR4、MyD88、NF-κB、hs-CRP、IL-6表达均明显上调；远程缺血后适应时大鼠海马CA1区TLR4、MyD88、hs-CRP、IL-6表达与单纯缺血再灌注损伤相比均明显下降；远程缺血后适应时大鼠海马CA1区NF-κB的表达与单纯缺血再灌注损伤时相比无明显变化。
[Abstract]:Objective To observe the changes in the expression of TLR4, MyD88, NF- kappa B, hs-CRP and IL-6 in the hippocampus CA1 region of rats after remote ischemic adaptation, and to explore the protective effect of TLR4-MyD88 signal transduction pathway in the long range ischemic adaptation of rats.
Methods 172 healthy male SD rats (weight 250g~300g) were randomly divided into model group (n=24), the experimental group, the remote ischemic postconditioning group (n=24), and the sham operation group (n=24). Each group was divided into 12h, 48h, 24h, 72h4 time point group (n=6).2 application improved Longa method, the rat model of local cerebral ischemia reperfusion was established. The group was reperfusion after MCAO2h, and the hippocampal pathological changes were observed immediately after the remote ischemic postconditioning.3HE staining. The immunohistochemical staining was used to detect the TLR4, MyD88, NF- kappa B, hs-CRP, IL-6 in the hippocampus CA1 region, and the.4 tissue section was taken by OLYMPUS camera microscope (x 200), and the Motic digital medical image was used. The data obtained by the analysis of.5 were analyzed by SPSS17.0 statistical software, and the measurement data were expressed with mean standard deviation (x + s), and the single factor variance analysis was done. The difference was statistically significant with the difference of P < 0.05. The correlation coefficient of each detection factor was analyzed with the correlation coefficient of Pearson, and the correlation of R 0 was found.
Results the hippocampal structure of the rats in the 1HE staining sham operation group was clear, the morphology of the neurons in the CA1 area was normal, the arrangement of the neurons was neatly arranged, the hippocampal structure in the model group was loose, the neuron loss in the CA1 area was obvious, and the cell body was narrowed, the nuclear condensation was deep, and the cell body swelled, the vacuoles were formed in the cytoplasm and the small necrotic area in the cytoplasm. In the experimental group, cell loss and cell swelling were significantly reduced compared with the model group, and the cell morphology improved.
2 the expression level of TLR4 in the hippocampal CA1 area in the sham operation group was lower. The expression level of TLR4 in the model group and the experimental group was significantly higher than that in the sham group (P < 0.05). The positive expression of TLR4 in the experimental group was lower than the model group before 72h (P < 0.05), but the 72h was slightly higher than the model group, but there was no statistical significance (P > 0.05) in the CA1 region of the hippocampus in the.3 sham operation group, at every time. The expression level of point MyD88 was lower. The expression of MyD88 in the model group and the experimental group was significantly higher (P < 0.05). The positive expression of MyD88 in the experimental group was lower than that in the model group (P < 0.05).4 sham operation group, and the expression level of NF- kappa B at all time points was lower. The expression of NF- kappa B in the model group and the experimental group was significantly higher than that in the sham operation group (P < 0.05). The positive expression of NF- kappa B in the test group was slightly lower than that in the model group at all time points, but there was no statistical significance (P > 0.05) in the CA1 region of the hippocampus of the.5 sham operation group, only a small amount of hs-CRP was expressed at each time point. The expression of hs-CRP in the model group and the experimental group was significantly higher than that in the sham group (P < 0.05), and the positive expression of hs-CRP in the experimental group was significantly lower than that in the model group at all time points (P < 0). .05). The CA1 area of the hippocampus in the sham operation group was only a small amount of expression at all time points. The expression of IL-6 in the model group and the experimental group was significantly higher than that in the sham operation group (P < 0.05). The positive expression of IL-6 in the experimental group was lower than the model group (P < 0.05) at all time points (P < 0.05). The expression of TLR4 and MyD88 was positively correlated in the model group, but the correlation was not significant (r=0.213, P=0.317). MyD88 was positively correlated with the expression level of NF- kappa B, but the correlation was not significant (r=0.302, P=0.151); NF- kappa B had a significant positive correlation with IL-6 expression (r=0.792, P < 0.001), and NF- kappa B was also positively correlated with the expression of IL-6.
Conclusion the expression of TLR4, MyD88, NF- - kappa B, hs-CRP, IL-6 in the hippocampus CA1 area of rats with ischemia-reperfusion injury was obviously up-regulated, and TLR4, MyD88, hs-CRP in hippocampus CA1 area of rats after remote ischemia, and the expression of IL-6 expression was significantly lower than that of simple ischemia reperfusion injury. There was no significant change in reperfusion injury.
【学位授予单位】：河北联合大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743

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