【摘要】：第一部分星形膠質(zhì)細(xì)胞與JAK-STAT通路在兔脊髓缺血再灌注損傷后變化規(guī)律的研究目的:觀察兔脊髓缺血再灌注后星形膠質(zhì)細(xì)胞(AS)、膠質(zhì)細(xì)胞源性神經(jīng)營養(yǎng)因子(GDNF)、腦源性神經(jīng)營養(yǎng)因子(BDNF)以及Janus激酶/信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活子(JAK-STAT)的表達規(guī)律。方法:采用胸主動脈阻斷法建立兔脊髓缺血(22min)/再灌注損傷模型。健康成年(36周)雄性新西蘭大白兔54只,采用計算機隨機數(shù)法隨機分為缺血再灌注Oh、1h、2h、3h、8h、24h、48h、72h組和假手術(shù)組(每組6只)共9組。缺血再灌注組分別于缺血再灌注后Oh、1h、2h、3h、8h、24h、48h和72h檢測缺血段脊髓組織中正常神經(jīng)元、凋亡神經(jīng)元以及GFAP(星形膠質(zhì)細(xì)胞特征性標(biāo)記物)、GDNF、BDNF、p-STAT3的表達水平。結(jié)果:正常神經(jīng)元數(shù)量隨著再灌注時間延長而減少;TUNEL染色結(jié)果顯示,脊髓缺血再灌注損傷后8h TUNEL陽性神經(jīng)元開始增多,24hTUNEL陽性神經(jīng)元最多。再灌注后3hGFAP表達水平開始增多,48h達到高峰;GDNF在再灌注后3h和72h各達到一次高峰;再灌注后24h BDNF表達水平最高;p-STAT3表達水平分別在再灌注后1h和48h到達高峰。假手術(shù)組:少見TUNEL陽性細(xì)胞,GFAP、GDNF、BDNF僅有少量表達,p-STAT3幾乎無表達。結(jié)論:脊髓缺血再灌注損傷可以誘發(fā)星形膠質(zhì)細(xì)胞激活,神經(jīng)營養(yǎng)因子GDNF、BDNF水平升高,JAK-STAT信號通路激活并呈現(xiàn)不同的表達規(guī)律。第二部分兔脊髓缺血再灌注后小膠質(zhì)細(xì)胞活化和炎性細(xì)胞因子變化的實驗研究目的:觀察兔脊髓缺血再灌注后小膠質(zhì)細(xì)胞活化及炎性因子IL-6、IL-10、核轉(zhuǎn)錄因子NF-κB的變化規(guī)律,為后處理干預(yù)時機提供理論依據(jù)。方法:采用胸主動脈球囊阻斷法建立兔脊髓缺血(22min)/再灌注損傷模型。健康成年(36周)雄性新西蘭大白兔54只,隨機分為假手術(shù)組S和缺血再灌注組C0、C1、C2、C3、C8、C24、C48、C72(每組6只)。缺血再灌注組分別于再灌注后Oh、1h、2h、3h、8h、24h、48h和72h應(yīng)用形態(tài)學(xué)、分子生物學(xué)等檢測方法檢測缺血段脊髓組織中正常神經(jīng)元、凋亡神經(jīng)元以及Iba-1(小膠質(zhì)細(xì)胞特征性標(biāo)記物)、IL-6、IL-10、NF-κB的表達水平。結(jié)果:正常神經(jīng)元數(shù)量隨再灌注時間延長而減少;脊髓缺血再灌注損傷后8h TUNEL陽性神經(jīng)元開始增多,24hTUNEL陽性神經(jīng)元達高峰。再灌注2hIba-1表達開始增多,8h達到高峰;NF-κB于再灌注3h開始增高,8h為表達高峰;IL-6和IL-10均在再灌注24h達到高峰。NF-κB、IL-6、IL-10的表達水平與Iba-1顯著正相關(guān)。結(jié)論:脊髓缺血再灌注后小膠質(zhì)細(xì)胞激活呈動態(tài)變化。NF-κB、IL-6、IL-10的表達水平與小膠質(zhì)細(xì)胞激活顯著正相關(guān),在小膠質(zhì)細(xì)胞激活前給予后處理可降低神經(jīng)元損傷。
[Abstract]:The study of changes of astrocytes and JAK-STAT Pathway after Spinal Cord Ischemia-reperfusion injury in Rabbits objective: to observe astrocytes (as), glial cell derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) after spinal cord ischemia-reperfusion injury in rabbits Expression of neurotrophic factor (BDNF) and Janus kinase / signal transduction and activator of transcription (JAK-STAT). Methods: rabbit spinal cord ischemia (22min) / reperfusion injury model was established by thoracic aortic occlusion. Fifty-four male New Zealand white rabbits (36 weeks old) were randomly divided into 9 groups (n = 6 in each group). The expressions of normal neurons, apoptotic neurons and GFAP (astrocytic characteristic marker) BDNF p-STAT3 in ischemic spinal cord were detected in the ischemia reperfusion group at 1 h, 3 h, 8 h, 24 h and 72 h after ischemia and reperfusion, respectively. Results: the number of normal neurons decreased with the prolongation of reperfusion time. The results of Tunel staining showed that the number of Tunel positive neurons began to increase at 8 h after spinal cord ischemia reperfusion injury, and the number of Tunel positive neurons increased most at 24 h after spinal cord ischemia reperfusion injury. The expression level of GFAP began to increase at 3 h after reperfusion and reached the peak at 48 h and reached the peak at 3 h and 72 h after reperfusion respectively, and the highest level of BDNF at 24 h after reperfusion reached the peak at 1 h and 48 h after reperfusion, respectively. In sham-operation group, few Tunel positive cells expressed BDNF and p-STAT3. Conclusion: spinal cord ischemia-reperfusion injury can induce astrocyte activation, and the level of neurotrophic factor GDNFN BDNF increases the activation of JAK-STAT signal pathway and presents different expression patterns. The second part of the experimental study on microglia activation and inflammatory cytokines after spinal cord ischemia reperfusion in rabbits objective: to observe the changes of microglia activation and inflammatory cytokine IL-10 and nuclear transcription factor NF- 魏 B after spinal cord ischemia reperfusion in rabbits. To provide theoretical basis for post-processing intervention timing. Methods: spinal cord ischemia (22min) / reperfusion injury model was established by balloon occlusion of thoracic aorta. Fifty-four male New Zealand white rabbits (36 weeks old) were randomly divided into sham-operated group (S) and ischemia reperfusion group (C0C _ 1C _ (2) C _ (2) C _ (3) C _ (3) C _ (24) C _ (48) C _ (72) (n = 6 in each group). The expressions of normal neurons, apoptotic neurons and Iba-1 (microglial characteristic marker) IL-10NF- 魏 B in ischemic spinal cord tissues were detected by morphological and molecular biology methods in the ischemia reperfusion group at 1 h, 2 h, 8 h and 24 h after reperfusion, respectively. Results: the number of normal neurons decreased with the prolongation of reperfusion time, and the number of Tunel positive neurons began to increase at 8 h after spinal cord ischemia-reperfusion injury and reached the peak at 24 h. The expression of Iba-1 began to increase at 2 h after reperfusion and reached the peak at 8 h. NF- 魏 B increased at 3 h after reperfusion. The expression peak of IL-6 and IL-10 reached the peak at 24 h after reperfusion. The expression level of IL-6 and IL-10 was positively correlated with Iba-1. Conclusion: the dynamic changes of microglia activation after spinal cord ischemia-reperfusion. The expression level of IL-6 / IL-10 is positively correlated with microglial activation, and post treatment before and after microglia activation can reduce neuronal injury.
【學(xué)位授予單位】：首都醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R651.2

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