本文選題：遠(yuǎn)程缺血后適應(yīng) + TLR4-MyD88�。� 參考：《河北聯(lián)合大學(xué)》2014年碩士論文

【摘要】：目的觀察大鼠遠(yuǎn)程缺血后適應(yīng)時海馬CA1區(qū)TLR4、MyD88、NF-κB、hs-CRP、IL-6表達(dá)變化，探討TLR4-MyD88信號轉(zhuǎn)導(dǎo)通路在大鼠遠(yuǎn)程缺血后適應(yīng)中的腦保護作用。 方法172只健康雄性SD大鼠（體重250g~300g）隨機分為模型組（n=24）；實驗組，即遠(yuǎn)程缺血后適應(yīng)組（n=24）；假手術(shù)組（n=24）。各組分為12h、48h、24h、72h4個時間點組（n=6）。2應(yīng)用改進的Longa法，建立大鼠大腦中動脈阻塞（MCAO）的局部腦缺血再灌注模型。實驗組于大鼠MCAO2h后再灌注，并即刻給予遠(yuǎn)程缺血后適應(yīng)處理。3HE染色觀察海馬病理學(xué)變化；使用免疫組織化學(xué)染色檢測海馬CA1區(qū)TLR4、MyD88、NF-κB、hs-CRP、IL-6的表達(dá)。4組織切片采用OLYMPUS攝像顯微鏡（×200）攝像，并應(yīng)用Motic Med6.0數(shù)碼醫(yī)學(xué)圖像分析系統(tǒng)進行分析。5所得數(shù)據(jù)均采用SPSS17.0統(tǒng)計軟件進行分析，計量資料用均數(shù)±標(biāo)準(zhǔn)差（x±s）表示，做單因素方差分析，以P＜0.05為差異有統(tǒng)計學(xué)意義。應(yīng)用Pearson相關(guān)系數(shù)分析各檢測因素相關(guān)性，r≠0為具有相關(guān)性。 結(jié)果1HE染色假手術(shù)組大鼠海馬結(jié)構(gòu)清晰，CA1區(qū)神經(jīng)元形態(tài)正常，排列整齊、層次分明；模型組大鼠海馬結(jié)構(gòu)松散，CA1區(qū)神經(jīng)元缺失明顯，可見較多神經(jīng)元胞體縮小、核固縮深染，也有胞體腫脹、胞漿內(nèi)形成空泡的神經(jīng)元以及小片狀壞死區(qū)域；實驗組神經(jīng)元缺失、細(xì)胞腫脹較模型組明顯減輕，細(xì)胞形態(tài)趨于改善。 2假手術(shù)組海馬CA1區(qū)，各時間點TLR4表達(dá)水平均較低。模型組與實驗組TLR4表達(dá)水平較假手術(shù)組明顯增高（P＜0.05）。實驗組TLR4陽性表達(dá)在72h前均低于模型組（P＜0.05），而72h時略高于模型組，但無統(tǒng)計學(xué)意義（P＞0.05）。3假手術(shù)組海馬CA1區(qū)，各時間點MyD88表達(dá)水平均較低。模型組與實驗組MyD88表達(dá)明顯增高（P＜0.05）。實驗組MyD88陽性表達(dá)在各時間點均低于模型組（P＜0.05）。4假手術(shù)組海馬CA1區(qū)，各時間點NF-κB表達(dá)水平均較低。模型組與實驗組NF-κB表達(dá)均較假手術(shù)組明顯增高（P＜0.05）。實驗組NF-κB陽性表達(dá)在各時間點均略低于模型組，但無統(tǒng)計學(xué)意義（P＞0.05）。5假手術(shù)組海馬CA1區(qū)，各時間點hs-CRP僅有少量表達(dá)，模型組與實驗組hs-CRP表達(dá)較假手術(shù)組明顯增高（P＜0.05），實驗組hs-CRP陽性表達(dá)在各時間點均明顯低于模型組（P＜0.05）。假手術(shù)組海馬CA1區(qū)，各時間點IL-6僅有少量表達(dá)，模型組與實驗組IL-6表達(dá)較假手術(shù)組明顯增高（P＜0.05），實驗組IL-6陽性表達(dá)在各時間點均低于模型組（P＜0.05）。6在模型組中TLR4與MyD88表達(dá)水平成正相關(guān)，但相關(guān)性并不顯著（r=0.213，P=0.317）；MyD88與NF-κB表達(dá)水平成正相關(guān)，，但相關(guān)性不顯著（r=0.302，P=0.151）；NF-κB與IL-6表達(dá)呈顯著正相關(guān)（r=0.792，P＜0.001）；NF-κB與hs-CRP表達(dá)也呈顯著正相關(guān)（r=0.767，P＜0.001）。 結(jié)論缺血再灌注損傷時大鼠海馬CA1區(qū)TLR4、MyD88、NF-κB、hs-CRP、IL-6表達(dá)均明顯上調(diào)；遠(yuǎn)程缺血后適應(yīng)時大鼠海馬CA1區(qū)TLR4、MyD88、hs-CRP、IL-6表達(dá)與單純?nèi)毖俟嘧p傷相比均明顯下降；遠(yuǎn)程缺血后適應(yīng)時大鼠海馬CA1區(qū)NF-κB的表達(dá)與單純?nèi)毖俟嘧p傷時相比無明顯變化。
[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.
【學(xué)位授予單位】：河北聯(lián)合大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743

【參考文獻】

相關(guān)期刊論文 前4條

1 ;Role of Toll-like receptors in regulatory functions of T and B cells[J];Chinese Science Bulletin;2008年08期

2 譚鋒;魏育濤;邱強;侯量;羅波;薛亞軍;;缺血后適應(yīng)的不同干預(yù)時間對家兔短期缺血再灌注心肌細(xì)胞凋亡影響[J];現(xiàn)代生物醫(yī)學(xué)進展;2010年21期

3 王貝娜;何龍泉;王玉;;短暫肢體缺血后適應(yīng)對急性腦梗死患者神經(jīng)功能恢復(fù)作用的臨床觀察[J];中風(fēng)與神經(jīng)疾病雜志;2012年05期

4 張興華;李春梅;馬曉靜;羅曼;;肢體與心肌缺血后處理對兔急性心肌缺血再灌注損傷的對比研究[J];中華醫(yī)學(xué)雜志;2006年12期

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