【摘要】：背景：肝臟缺血再灌注(ischemia-reperfusion, IR)損傷是臨床工作中常會遇見的問題,并且,隨著國家取消尸肝供體移植,心死亡后器官捐獻的(Donation after circulation death, DCD)肝臟已經(jīng)成為當今肝移植的主要來源,而DCD供肝的缺血再灌注損傷程度將會明顯影響移植肝的功能恢復,甚至會決定肝移植手術(shù)的成敗,因此如何有效地減輕肝臟IR損傷程度將是DCD供肝肝移植手術(shù)成功的關(guān)鍵。全反式維甲酸(all-trans retinoic acid, ATRA)是臨床上用來治療急性早幼粒白血病的常用藥,現(xiàn)在研究發(fā)現(xiàn)ATRA也具有減輕肝IR損傷,但是其具體作用機制還不清楚,本實驗旨在闡釋ATRA在肝IR損傷中的作用機制。方法：體內(nèi)實驗：采用70%肝IR損傷模型,分為空白組,IR組和ATRA預處理組,在相應時間點,采血后處死小鼠并取材,進行肝功能檢測,觀察肝臟病理并評分,并利用RT-PCR和蛋白印記等技術(shù)對相應指標進行檢測；體外實驗：采用過氧化氫(Hydrogen peroxide, H2O2)和脂多糖(1ipopolysaccharides, LPS)分別處理小鼠肝細胞FL83B和巨噬細胞RAW264.7,分組情況均為DMSO、 H2O2/LPS、ATRA預處理和LE540預處理組,在相應時間點,收集細胞后,再利用RT-PCR和蛋白印記等技術(shù)進行分析。結(jié)果：(1)IR組中,肝功能、肝臟組織病理評分均高于空白組且有統(tǒng)計學差異；(2) ATRA預處理組中,肝功能及肝臟組織病理評分均低于IR組,炎癥因子TNF-α、IL-6和IL-1β mRNA等水平也有明顯下降；(3)ATRA預處理組中,維甲酸受體α、Bcl-2、p-Akt、Beclin1和LC3Ⅱ/Ⅰ等蛋白水平均高于IR組,cleaved-caspase 3、 Foxol、 TLR4、NF-κB p65和p62等蛋白水平低于IR組；(4)在FL83B試驗中,ATRA預處理后使細胞凋亡減少,維甲酸受體α、Bc1-2、p-Akt、 Beclin1、LC3Ⅱ/Ⅰ等蛋白水平均高于H202組,cleaved-caspase 3、Foxo1、p62等蛋白水平低于H202組,而LE540預處理后損傷明顯加重；(5)在RAW264.7試驗中,ATRA預處理組中,炎癥因子TNF-α、IL-6和IL-1β mRNA等水平明顯下降,維甲酸受體a、p-Akt、Beclinl和LC3Ⅱ/LC3Ⅰ等蛋白水平均高于LPS組,Foxo1、TLR4、NF-κB p65和p62等蛋白水平低于LPS組,而LE540預處理后,卻進一步加重了炎癥反應。結(jié)論：(1)70%肝IR損傷模型建立成功；(2) ATRA可以減輕肝臟R損傷：①減少細胞凋亡,②調(diào)節(jié)先天免疫反應；(3) ATRA的保護作用可能與激活維甲酸受體α有關(guān)。
[Abstract]:Background: liver ischemia-reperfusion (ischemia-reperfusion, IR) injury is a common problem in clinical work. With the cancellation of cadaveric liver donor transplantation, (Donation after circulation death, DCD) liver donated after cardiac death has become the main source of liver transplantation. However, the degree of ischemia-reperfusion injury of DCD donor liver will significantly affect the functional recovery of the transplanted liver and even determine the success or failure of liver transplantation. Therefore, how to effectively reduce the degree of liver IR injury will be the key to the success of DCD donor liver transplantation. All trans retinoic acid (all-trans retinoic acid, ATRA) is a common drug used in the treatment of acute promyelocytic leukemia. Now it has been found that ATRA can also attenuate liver IR damage, but its mechanism is not clear. The purpose of this study was to elucidate the role of ATRA in liver IR injury. Methods: in vivo experiment, 70% liver IR injury model was used and divided into two groups: control group (IR group) and ATRA pretreatment group (P < 0.05). The mice were sacrificed at the corresponding time point after blood collection, and the liver function was detected, liver pathology was observed and evaluated. RT-PCR and protein imprinting techniques were used to detect the corresponding indexes. In vitro experiments, FL83B and RAW264.7, of mouse hepatocytes were treated with hydrogen peroxide (Hydrogen peroxide, H2O2 and lipopolysaccharide (LPS) respectively as DMSO, H _ 2O _ 2 / LPSN _ (ATRA) pretreatment group and LE540 pretreatment group. Cells were collected at the corresponding time points, and then analyzed by RT-PCR and protein imprinting techniques. Results: (1) in IR group, liver function and liver histopathological score were significantly higher than those in blank group, (2) in ATRA preconditioning group, liver function and liver histopathological score were lower than those in IR group, and the levels of inflammatory factor TNF- 偽, IL-6 and IL-1 尾 mRNA were significantly decreased. (3) in the ATRA pretreatment group, the protein levels of retinoic acid receptor 偽 -Bcl-2p-Akton-Beclin1 and LC3 鈪，

本文編號：2225434