本文關鍵詞：大鼠肺移植供肺缺血再灌注的自噬水平變化　出處：《南昌大學》2015年碩士論文　論文類型：學位論文

  更多相關文章： 自噬 肺移植 缺血再灌注損傷 大鼠

【摘要】：目的:實驗通過建立大鼠原位肺移植模型,利用實驗動物學及分子生物學的技術(shù),闡明大鼠肺移植供肺灌注、冷缺血、再灌注后自噬水平變化,為調(diào)控自噬水平,減輕肺移植缺血再灌注損傷提供理論依據(jù)。方法:1、分組并建立模型:SPF級雄性SD大鼠,共60只,質(zhì)量300~350g。隨機挑選12只作單純灌注冷缺血處理,其中6只作為單純灌注冷缺血0h對照,另外6只作單純灌注冷缺血后保存6h處理;其余48只分為供、受體,受體略大于供體,供、受體間質(zhì)量差不超過20g,進行左肺移植手術(shù)。本實驗共分五組:(1)CON組即灌注后,冷缺血保存0h組;(2)ISC6組即灌注后,冷缺血保存6h組;(3)REP2組即灌注后,先冷缺血保存6h,移植后再灌注2h組;(4)REP6組即灌注后,先冷缺血保存6h,移植后再灌注6h組;(5)REP12組即灌注后,先冷缺血保存6h,移植后再灌注12h組。其中CON,ISC6組各6例,REP2,REP6,REP12組各移植6對大鼠。2、分子生物學檢測:免疫熒光雙染LC3,LAMP-2,檢測自噬流,Western blot檢測LC3-II/LC3-I。結(jié)果:1、成功建立了一種經(jīng)過改良了肺移植技術(shù)的大鼠原位左肺移植模型;2、通過免疫熒光雙染LC3,LAMP-2,發(fā)現(xiàn):CON組自噬流最低,ISC6組自噬流稍升高,REP2,REP6組自噬流顯著升高,其中REP6組自噬流最高,REP12組自噬流降低。3、通過Western blot檢測LC3蛋白表達發(fā)現(xiàn):CON組LC3蛋白表達最低,ISC6組LC3蛋白表達稍升高,REP2,REP6組LC3蛋白表達顯著升高,其中REP6組LC3蛋白表達最高,REP12組LC3蛋白表達降低。結(jié)論:大鼠肺移植供肺灌注、冷缺血6h后自噬水平開始增高,再灌注2h的自噬水平明顯增高,再灌注6h達到高峰,再灌注12h后自噬減輕。
[Abstract]:Objective: through the establishment of experimental model of orthotopic lung transplantation in rats, the use of laboratory animal science and molecular biology technology, clarify the rat lung transplantation for pulmonary perfusion and cold ischemia after reperfusion, the level of autophagy changes, for the regulation of autophagy, alleviate the ischemia reperfusion injury in lung transplantation and provide a theoretical basis for the. Methods: 1, group and model: grade SPF male SD rats, with a total of 60 rats, with a mass of 300~350g. 12 randomly selected only for simple perfusion cold ischemia treatment, including 6 as simple perfusion of cold ischemia for 0h control, the other 6 only for simple perfusion cold ischemia preserved after 6h treatment; the remaining 48 were divided into donor and receptor, receptor is slightly larger than the donor between donor and recipient of poor quality does not exceed 20g, left lung transplantation surgery. The experiments were divided into five groups: (1) CON group after reperfusion, cold ischemia 0h group; (2) ISC6 group after reperfusion, cold ischemia 6h group; (3) REP2 group after reperfusion, first cold ischemia 6h, reperfusion after transplantation in 2H group; REP6 group (4) the first cold ischemia after reperfusion, 6h after transplantation, reperfusion 6h group; (5) REP12 group after reperfusion, first cold ischemia 6h, reperfusion after transplantation in 12h group. 6 cases in each group of CON, group ISC6, REP2, REP6, and REP12 were transplanted in 6 pairs of rats. 2, molecular biological detection: immunofluorescence double staining LC3, LAMP-2, detection of autophagic flow, Western blot detection of LC3-II/LC3-I. Results: 1, successfully established a modified rat model of orthotopic left lung transplantation and lung transplantation technology; 2, by double immunofluorescence staining LC3, LAMP-2, CON group found that autophagy flow is the lowest, ISC6 group REP2, autophagy flux was slightly higher in REP6 group significantly increased the flow of autophagy, autophagy group REP6 flow the highest REP12 group decreased autophagy flow. 3, the expression of LC3 protein was detected by Western blot. It was found that the expression of LC3 protein was the lowest in group CON, the expression of LC3 protein increased slightly in ISC6 group, and the expression of LC3 protein in REP2 and REP6 group increased significantly, among which the expression of LC3 protein in the REP6 group was the highest and the expression of group C was decreased. Conclusion: autophagy level is increased after lung transplantation and cold ischemia 6h in rat lung transplantation. The autophagy level of 2h after reperfusion is significantly higher than that of reperfusion, and the peak of 6h is reached after reperfusion, and autophagy is alleviated after 12h reperfusion.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R655.3

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本文編號：1338597