【摘要】：目的:對高脂血癥大鼠施行局灶性腦缺血再灌注術(shù),觀察缺血后處理對缺血側(cè)腦組織中STAT-3、IL-6及凋亡細(xì)胞的動態(tài)表達(dá),探討缺血后處理對高脂血癥大鼠腦缺血再灌注損傷的保護(hù)作用及可能機(jī)制。方法:將115只SD大鼠隨機(jī)分為兩組,其中一組用普通飼料喂養(yǎng)(10只),另一組用高脂飼料喂養(yǎng)(105只),于28天后,空腹心臟采血檢測兩組大鼠血清中的血脂指標(biāo),分別為血清總膽固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C),高密度脂蛋白(HDL-C),兩組含量比較,若存在明顯差異(P0.05),表明高血脂血癥大鼠造模成功。隨機(jī)將造模成功的高脂血癥大鼠分為假手術(shù)組(sham組)35只、缺血再灌注組(I/R組)35只、缺血后處理組(IP組)35只。I/R組與IP組動物模型的制作均使用改良后的線栓法,并在大鼠腦缺血2小時(shí),進(jìn)行再灌注,術(shù)后按時(shí)間點(diǎn)的不同將30只大鼠平均分為6組,各時(shí)間點(diǎn)的設(shè)置分別是3h、6h、12h、24h、48h、72h,IP組于再灌注前給予灌注15s/缺血15s,反復(fù)3個(gè)循環(huán),sham組僅分離頸動脈后縫合切口。sham組于手術(shù)后24小時(shí)使用5只大鼠取腦組織行TTC染色,IP組與I/R組于再灌注后24小時(shí)使用5只大鼠行TTC染色,計(jì)算腦梗死體積。各時(shí)間點(diǎn)神經(jīng)功能缺損情況通過神經(jīng)功能缺損評分判斷,不同時(shí)間點(diǎn)腦梗死組織的病理改變通過HE染色觀察,神經(jīng)細(xì)胞凋亡的數(shù)量采用Tunel法檢測,STAT-3和IL-6的表達(dá)用免疫組化(Envision法)檢測。結(jié)果:1.高脂血癥大鼠模型:高脂飼料喂養(yǎng)組大鼠血清中TC、TG、LDL-C、HDL-C的含量明顯高于普通飼料喂養(yǎng)組(P0.05);2.神經(jīng)功能缺損評分:sham組大鼠沒有出現(xiàn)明顯的神經(jīng)功能缺損癥狀,I/R組于再灌注不同時(shí)間點(diǎn)出現(xiàn)不同程度神經(jīng)功能障礙,以24小時(shí)評分最低,與其他時(shí)間點(diǎn)比較差異有統(tǒng)計(jì)學(xué)意義,IP組在12小時(shí)以后神經(jīng)功能評分顯著高于相同時(shí)間點(diǎn)I/R組(P0.05);3.TTC染色計(jì)算腦梗死體積:sham組術(shù)后24小時(shí),TTC染色腦組織為均一的紅色,腦組織形態(tài)正常,無明顯蒼白和水腫;I/R組和IP組術(shù)后24小時(shí)的腦組織內(nèi)可觀察到線栓側(cè)腦組織顏色蒼白,體積大于無線栓側(cè)的梗死腦組織,I/R組梗死體積較IP組大(P0.05);4.神經(jīng)細(xì)胞凋亡數(shù)量及IL-6、STAT-3免疫陽性細(xì)胞的表達(dá):sham組的腦組織經(jīng)過免疫組化染色后在鏡下未見STAT-3免疫陽性細(xì)胞,偶可見少量IL-6免疫陽性細(xì)胞表達(dá);I/R組腦缺血再灌注后3h,STAT-3開始少量表達(dá),IL-6表達(dá)增多,24h達(dá)到高峰后開始下降,在72h仍有高表達(dá);IP組STAT-3和IL-6的表達(dá)與I/R組變化趨勢相似,但數(shù)量顯著減少(P0.05),凋亡細(xì)胞變化與STAT-3、IL-6基本一致。結(jié)論:1.在建立高脂血癥大鼠模型基礎(chǔ)上制作一側(cè)大腦中動脈腦缺血再灌注模型,能更好的模擬人體腦梗死發(fā)病模式,使所檢測出來的各項(xiàng)指標(biāo)更加具有說服力;2.腦缺血再灌注后可使大鼠出現(xiàn)行為學(xué)及形態(tài)學(xué)方面的損傷,使IL-6、凋亡細(xì)胞及STAT-3表達(dá)增多;3.缺血后處理可能的腦保護(hù)機(jī)制是通過抑制IL-6的表達(dá),進(jìn)而減少JAK2/STAT3通路的激活,減輕凋亡程度。
[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.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R743.3

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