本文選題：異丙酚　切入點：缺血再灌注損傷　出處：《天津醫(yī)科大學》2017年碩士論文

【摘要】：目的:臨床上肝移植或其他復雜肝臟外科手術(shù)中必然會涉及肝缺血再灌注這一病理過程,肝缺血再灌注不僅能夠損害肝臟本身,還可能誘發(fā)遠隔臟器損傷,嚴重影響患者生存預后。如何保護遠隔臟器,并且減少再灌注后損傷的發(fā)生,已成為近年來研究的熱點。本研究主要研究大鼠肝冷缺血再灌注過程中JAK/STAT信號通路相關(guān)蛋白的表達情況,并探討異丙酚能否通過調(diào)節(jié)JAK/STAT信號通路對肝冷缺血再灌注后腎損傷產(chǎn)生影響,以揭示肝冷缺血再灌注誘發(fā)腎損傷過程中的可能機制,為肝缺血再灌注后遠隔臟器損傷的防治提供新思路。方法:SD成年雄性健康大鼠,周齡8~10周,體重220~250 g,隨機分為4組(n=8):假手術(shù)組(Sham組);肝冷缺血再灌注改良模型組(I/R組);異丙酚組(Pro組);JAK2激酶抑制劑AG490組(AG490組)。Pro組大鼠行右側(cè)股靜脈置管連接異丙酚微量輸液泵后,于再灌注前5 min泵注異丙酚20 mg·kg-1·h-1持續(xù)30 min。AG490組于建立模型前30 min腹腔注射AG490 10 mg/kg。再灌注6 h后,經(jīng)下腔靜脈采集血樣后處死大鼠,使用全自動生化分析儀檢測大鼠血清BUN、Cr含量變化,ELISA法測定TNF-α和IL-6的濃度。取腎組織標本,測定腎組織MDA含量及SOD水平。HE染色與TUNEL法觀察腎組織病理學變化與細胞凋亡情況,并對腎小管損傷程度進行評分、計算凋亡指數(shù)。免疫組織化學法檢測腎組織細胞內(nèi)Cleaved Caspase-3的表達情況;Western blot法檢測p-JAK2、p-STAT1和p-STAT3的蛋白表達水平。應用SPSS21.0統(tǒng)計軟件分析實驗結(jié)果,P0.05具有統(tǒng)計學差異。結(jié)果:與Sham組比較,I/R組大鼠腎功能衰退明顯,血清BUN和Cr濃度明顯升高,炎癥反應及氧化應激增強,血清TNF-α、IL-6濃度和腎組織MDA含量明顯升高,SOD活性降低(P0.05)。光鏡下可見I/R組大鼠多數(shù)腎小管上皮細胞水腫、空泡變性,腎小管擴張,管腔狹窄,炎癥細胞浸潤,間質(zhì)擴張淤血,腎小管損傷評分較Sham組顯著升高(P0.05)。Sham組腎組織細胞TUNEL標記的凋亡細胞較少,而I/R組腎組織凋亡細胞顯著增多,凋亡指數(shù)明顯升高,且腎組織細胞中Cleaved Caspase-3的表達水平升高(P0.05)。I/R組大鼠腎組織p-JAK2、p-STAT1和p-STAT3的蛋白水平顯著上調(diào)(P0.05)。與I/R組比較,Pro組及AG490組大鼠腎功能明顯改善,血清BUN、Cr濃度明顯降低,炎癥反應減輕,血清TNF-α、IL-6的濃度顯著降低,氧化應激緩解,腎組織MDA含量降低且SOD活性升高(P0.05)。Pro組與AG490組大鼠腎臟病理損傷較I/R組明顯減輕,腎小管損傷評分降低,TUNEL標記的腎組織凋亡細胞數(shù)量減少,凋亡指數(shù)顯著降低,且腎組織中Cleaved Caspase-3的表達明顯減少(P0.05)。與此同時,Pro組及AG490組p-JAK2、p-STAT1和p-STAT3蛋白水平與I/R組比較顯著下調(diào)(P0.05)。Pro組與AG490組各項指標差異無統(tǒng)計學意義。結(jié)論:(1)肝冷缺血再灌注可導致大鼠腎組織損傷,其機制與炎癥反應、氧化應激和細胞凋亡等多種因素有關(guān);(2)肝冷缺血再灌注能夠激活JAK/STAT信號通路,抑制此通路可顯著減輕肝冷缺血再灌注導致的腎損傷;(3)異丙酚能夠減輕大鼠肝冷缺血再灌注導致的腎損傷,機制可能與抑制JAK/STAT信號通路從而減輕氧化應激、緩解炎癥反應,抑制腎組織細胞凋亡有關(guān)。
[Abstract]:Objective: the clinical liver transplantation or other complex liver surgery must be involved in hepatic ischemia reperfusion of the pathological process, hepatic ischemia reperfusion can damage the liver itself, may also induce distant organ damage, seriously affect the prognosis. How to protect the remote organs, and reduce the occurrence of injury after reperfusion has become the research hotspot in recent years. The expression of JAK/STAT signaling pathway related protein in the research of cold ischemia reperfusion in rat liver, and to explore the effect of propofol can produce kidney injury by regulating JAK/STAT signal pathway on liver ischemia reperfusion injury, in order to reveal the possible mechanism of liver cold ischemia and reperfusion induced renal injury in the process and provide new ideas for the prevention of remote organ injury after liver ischemia reperfusion in rats. Methods: SD rats, 8~10 weeks, weighing 220~250 g, were randomly divided into The 4 group (n=8): sham operation group (group Sham); liver cold ischemia reperfusion model group (group I/R); Propofol group (Pro group); JAK2 kinase inhibitor AG490 group (group AG490).Pro group rats underwent right femoral vein catheter connected with propofol infusion pump, before reperfusion 5 min pump propofol 20 mg - kg-1 - H-1 for 30 min.AG490 group to model 30 min before intraperitoneal injection of AG490 10 mg/kg. 6 h after reperfusion, blood samples were collected from the inferior vena cava after rats were sacrificed, using automatic biochemical analyzer to detect the serum BUN, Cr content, determination of TNF- alpha and IL-6 ELISA method. Renal tissue samples, determination of renal tissue MDA content and SOD level of.HE and TUNEL staining method was used to observe the pathological changes and cell apoptosis, and the score of the degree of renal tubular injury, apoptosis index is calculated. The expression of immunohistochemical detection of kidney cells Cleaved Caspase-3 P-JAK2 Western blot; detection method, the expression level of p-STAT1 and p-STAT3 protein. The results were analyzed by SPSS21.0 statistical software, P0.05 has a statistically significant difference. Results: compared with Sham group, the renal function of I/R rats decline significantly, serum BUN and Cr were significantly increased, inflammatory reaction and oxidative stress increased, serum alpha TNF- the content of MDA, IL-6 concentration and renal tissue were significantly increased, SOD activity decreased (P0.05). Under visible edema, rats in the I/R group the majority of renal tubular epithelial cells light vacuolar degeneration, tubular dilatation, stenosis, inflammatory cell infiltration, interstitial congestion expansion, renal tubular injury score were significantly higher than that of Sham group (P0.05) a few apoptotic cells in renal tissues in.Sham group of TUNEL labeled cells, and apoptosis in the I/R group of renal tissue increased significantly, apoptosis index was significantly increased, and the expression of Cleaved in renal tissue cells in water Ping Shenggao Caspase-3 (P0.05) in kidney of rats in.I/R group P-JAK2, p-STAT1 protein level and p-STAT3 significantly increased (P0.05). Compared with I/R group, AG490 group and renal function of rats in group Pro were significantly improved, serum BUN, Cr concentrations were significantly decreased, inflammatory reaction, serum TNF-, IL-6 concentrations were significantly decreased, alleviate oxidative stress, MDA content of renal tissue decreased the increased activity of SOD (P0.05).Pro group and AG490 group rat kidney pathological damage significantly reduced compared with group I/R, renal tubular injury score decreased, TUNEL marked the number of apoptotic cells in renal tissue decreased, the apoptosis index decreased significantly, and the expression of Cleaved in renal tissue Caspase-3 decreased significantly (P0.05). At the same time, Pro group and AG490 group. P-JAK2, p-STAT1 and p-STAT3 protein levels compared with I/R group significantly decreased (P0.05) there was no significant difference between.Pro group and AG490 group index. Conclusion: (1) liver cold ischemia reperfusion can lead to kidney injury in rats, and the mechanism of anti inflammation Be related to various factors of oxidative stress and apoptosis; (2) liver cold ischemia reperfusion can activate JAK/STAT signaling pathway, inhibition of renal injury in this pathway can significantly reduce the liver cold ischemia reperfusion induced renal injury; (3) propofol can relieve the liver cold ischemia reperfusion in rats caused by the mechanism might be related to the inhibition the JAK/STAT signaling pathway to reduce oxidative stress, relieve inflammation, inhibit the apoptosis of renal tissue.

【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R614

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本文編號：1723434