發(fā)布時間：2018-05-20 00:37

  本文選題：TLR3 + 七氟烷　； 參考：《大連醫(yī)科大學》2014年碩士論文

【摘要】：目的：探討TLR3信號通路在大鼠七氟烷預處理減輕體外循環(huán)腦損傷中的作用，及其可能的分子機制。 方法：雄性SD大鼠64只。隨機分為3組：假手術組（H組，n=8），CPB組（C組，n=24）和七氟烷預處理組（S組，n=32）。假手術組(H組)：氣管插管機械通氣，血管穿刺置管，不進行體外循環(huán)轉(zhuǎn)流。CPB組（C組）：在右股動脈和右側(cè)頸內(nèi)靜脈穿刺置管，建立無血預充大鼠體外循環(huán)模型。七氟烷預處理(S組)：2.4%七氟烷預處理1h，建立體外循環(huán)模型。C組和S組進行體外循環(huán)1h。選擇麻醉后體外循環(huán)前（T0）、體外循環(huán)30分鐘(T1)、體外循環(huán)1h(T2)、體外循環(huán)后1h(T3)、體外循環(huán)后2h(T4)和體外循環(huán)后3h(T5)，采集血樣，記錄大鼠生命體征。C組和S組在T0、T1、T3和T5時，取8只大鼠處死取腦組織樣本。使用ELISA酶聯(lián)免疫法檢測血清S100-β、IL-6和IFNβ濃度變化；ELISA酶聯(lián)免疫法檢測左側(cè)海馬組織TLR3、TRIF表達情況；采用Western-Blot法檢測左側(cè)海馬組織TLR3表達情況；選取右側(cè)海馬，使用TUNEL法檢測神經(jīng)元調(diào)亡情況。 結(jié)果： 三組大鼠各時間點平均動脈壓（MAP）、心率（HR）、紅細胞壓積（Hct）、直腸溫、pH值、PaCO2、PaO2，BE值、K+組間比較差異無統(tǒng)計學意義（P0.05）。僅體外循環(huán)期間MAP、HR和Hct呈明顯降低（與T0比較，P0.05）。 對比H組，C組與S組血清S100-β和IL-6均升高（P0.05）。C組與S組血清S100-β和IL-6濃度轉(zhuǎn)流期間明顯升高，轉(zhuǎn)流結(jié)束后逐漸降低（與T0比較，P0.05）；對比C組T1~T5時，S組血清S100-β和IL-6濃度出現(xiàn)下降（P0.05）。 C組與S組在T1~T5時，血清IFNβ濃度比H組明顯升高（P0.05）；與C組對比,除外T0時S組各時間點血清IFNβ濃度均升高（P0.05）。 對比H組，，C組與S組TLR3和TRIF蛋白水平均出現(xiàn)明顯升高（P0.05）。C組和S組T2時TLR3和TRIF蛋白水平明顯升高，T3時TLR3和TRIF蛋白水平降低（與T0比較，P0.05）。對比H組T1、T3、T5時，C組與S組TLR3和TRIF蛋白水平均升高（P0.05）。 Western blot結(jié)果顯示，C組和S組TLR3蛋白表達量比H組顯著升高（與T0比較，P0.05）；S組和C組相比，TLR3表達量明顯升高（P0.05）。 TUNEL法檢測神經(jīng)元調(diào)亡顯示，H組海馬神經(jīng)元凋亡神經(jīng)元數(shù)目極少。對比H組，C組及S組海馬凋亡神經(jīng)元明顯增加（P0.05）；S組海馬凋亡神經(jīng)元數(shù)量比C組明顯降低（P0.05）。 結(jié)論： 1.2.4%七氟烷預處理對CPB腦損傷具有一定保護作用。 2.七氟烷預處理可通過影響TLR3信號通路，使其表達上調(diào)，減輕CPB炎癥反應造成的腦損傷進而發(fā)揮腦保護作用。 3. TLR3信號通路參與七氟烷預處理腦保護作用，可能是通過七氟烷預先激活TLR3，使下游TRIF表達上調(diào)，抑制促炎性因子IL-6的表達，上調(diào)抗炎性因子IFNβ的表達，從而對抗炎癥損害起到腦保護作用。
[Abstract]:Aim: to investigate the role of TLR3 signaling pathway in the protective effect of sevoflurane preconditioning on brain injury after cardiopulmonary bypass (CPB) and its possible molecular mechanism. Methods: 64 male SD rats were used. They were randomly divided into 3 groups: sham operation group (group H) and group C (group C) and group S (group S). Group H: mechanical ventilation of tracheal intubation, catheterization without cardiopulmonary bypass. Group C: puncture of right femoral artery and right internal jugular vein to establish rat model of cardiopulmonary bypass without blood supply. The model of cardiopulmonary bypass (CPB) was established in group C and group S for 1 hour after preconditioning with sevoflurane for 1 hour in group S: 2. 4% sevoflurane. Blood samples were collected before cardiopulmonary bypass (CPB), 30 minutes after cardiopulmonary bypass (CPB), 1 hour after cardiopulmonary bypass (CPB), 2 hours after cardiopulmonary bypass (CPB), and 3 hours after cardiopulmonary bypass (CPB). Blood samples were collected, and the vital signs of rats were recorded at T _ 0T _ 1T _ 3 and T _ 5 in group S and group C. Eight rats were killed and brain tissue samples were taken. The changes of serum S100- 尾 -IL-6 and IFN 尾 concentration were detected by ELISA enzyme-linked immunosorbent assay (ELISA). The expression of TLR3TIF in left hippocampus was detected by Elisa, TLR3 expression in left hippocampal tissue was detected by Western-Blot method, and the right hippocampus was selected. The apoptosis of neurons was detected by TUNEL method. Results: There was no significant difference in mean arterial pressure (MAPV), heart rate (HRT), HCT (HCT), rectal temperature and pH value (Paco _ 2 / Pao _ 2 ~ (2 +) be) between the three groups (P 0.05). During cardiopulmonary bypass (CPB), MAPHR and Hct decreased significantly (compared with T0, P 0.05). The levels of serum S100- 尾 and IL-6 in group C and group S were significantly higher than those in group C and group S during bypass, and gradually decreased after bypass (compared with T0, P 0.05), and serum S100- 尾 and IL-6 in group C and group S were decreased (P0.05A) when compared with that in group C and group S (P < 0.05). The serum S100- 尾 and IL-6 levels in group C and S were significantly lower than those in group C and group S (P < 0.05), and the levels of serum S100- 尾 and IL-6 in group C were significantly lower than those in group S. The concentration of serum IFN 尾 in group C and group S was significantly higher than that in group H at T1~T5, and that in group S was significantly higher than that in group C at all time points except T0, with the exception of T0, the concentration of serum IFN 尾 in group S was significantly higher than that in group H (P 0.05). Compared with group H and group S, the protein levels of TLR3 and TRIF in group C and group S were significantly increased. The protein levels of TLR3 and TRIF in group C and group S were significantly higher than those in group T 3. The protein levels of TLR3 and TRIF in group C were significantly higher than those in group S (P 0.05 compared with T0). The protein levels of TLR3 and TRIF in group C and group S were significantly higher than those in group T _ 1, T _ 3 and T _ 5 in H group (P 0.05). The results of Western blot showed that the expression of TLR3 protein in group C and group S was significantly higher than that in group H (compared with T0, the expression of TLR3 in group S and group C was significantly higher than that in group T 0. The number of apoptotic neurons in hippocampal neurons in H group was very small by TUNEL assay. Compared with group C and group S, the number of apoptotic neurons in hippocampus of group H and group S was significantly increased than that of group C, and the number of apoptotic neurons in group S was significantly lower than that in group C. Conclusion: 1.2.4% sevoflurane pretreatment had a protective effect on CPB brain injury. 2. Sevoflurane preconditioning can up-regulate the expression of TLR3 signal pathway and attenuate the brain injury caused by CPB inflammation and then play a protective role in the brain. 3. The TLR3 signaling pathway is involved in the protective effect of sevoflurane preconditioning. It may be that sevoflurane preactivates TLR3, up-regulates the expression of downstream TRIF, inhibits the expression of pro-inflammatory factor IL-6 and up-regulates the expression of anti-inflammatory factor IFN 尾. Thus, anti-inflammatory damage plays a brain protection role.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R614

【參考文獻】

相關期刊論文 前6條

1 楊寶會;趙立明;曲向林;盧立春;;七氟醚在體外循環(huán)冠狀動脈旁路移植術中的心肌保護作用[J];齊齊哈爾醫(yī)學院學報;2011年06期

2 陳燕;陳文棟;劉曼;;七氟醚麻醉對體外循環(huán)心臟手術患者圍術期血清HO-1水平的影響[J];山東醫(yī)藥;2011年22期

3 梅波;賴應龍;;體外循環(huán)腦損傷及腦保護研究進展[J];中國體外循環(huán)雜志;2010年02期

4 韓文冬;張麗娟;;七氟醚后處理對體外循環(huán)心內(nèi)直視手術患者圍手術期腦保護作用[J];中國現(xiàn)代醫(yī)學雜志;2012年22期

5 左蕾;李琪英;劉丹彥;;不同濃度七氟醚對老年大鼠認知功能的影響[J];重慶醫(yī)科大學學報;2013年03期

6 鄭賢軍;姜金玉;孔高茵;;七氟醚預處理對體外循環(huán)心臟瓣膜置換術患者血清S100β蛋白濃度的影響[J];中國醫(yī)藥指南;2013年22期

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