急性高糖对大鼠肾缺血再灌注损伤及右美托嘧啶干预的影响研究
发布时间：2018-10-26 21:15
【摘要】：急性高糖是指由于某些原因导致机体血糖水平显著升高的现象,当前包括肾脏移植在内的临床研究发现围术期高血糖水平与存在肾脏缺血再灌注患者的预后有着密切关系,但对其中的具体影响机制仍不清楚。本研究通过建立正常SD大鼠肾脏I/R急性高糖模型,评价在该模型下急性高糖对肾I/R损伤的影响和探讨急性高糖对肾I/R损伤可能机制,此外对右美托嘧啶在该急性高糖模型中的影响及可能机制进行探讨。第一部分急性高糖对大鼠肾缺血再灌注损伤的影响目的：探讨缺血前急性高糖对正常大鼠肾I/R的影响。方法：通过建立正常SD大鼠肾I/R急性高糖模型,与单纯肾I/R比较,采用形态学和功能学两方面研究缺血前急性高糖对正常大鼠肾I/R的影响。结果：缺血前急性高糖可以加重肾缺血再灌注损伤,再灌注后24h的Cr和BUN较非高糖I/R组明显升高。结论：急性高糖可以加重正常大鼠肾脏I/R损伤。第二部分急性高糖环境下大鼠肾缺血再灌注损伤的免疫炎症反应研究目的：探讨缺血前急性高糖对非糖尿病大鼠肾I/R免疫炎症反应的影响。方法：采用分光光度法使用髓过氧化物酶(MPO)测定试剂盒测定肾组织匀浆中MPO活性。采用免疫组织化学、RT-PCR测定IL-1β, interferon-γ (IFN-γ),采用免疫组织化学、RT-PCR、Western blot法测定肾组织TLR4,TLR2,高迁移率族蛋白B1 (high-mobility group box 1,HMGB1),Cyclooxygenase 2(Cox-2)的表达。结果：急性高糖增加缺血再灌注后肾组织MPO活性,增加IL-1β, IFN-γ, TLR4, TLR2,HMGB1,Cox-2的表达水平。结论：急性高糖加重肾缺血及再灌注所引发的免疫炎性损伤。第三部分急性高糖对大鼠肾缺血再灌注损伤凋亡的影响以及右美托嘧啶预处理的影响目的：急性高糖对大鼠肾缺血再灌注损伤凋亡的影响以及右美托嘧啶预处理的影响方法：非糖尿病SD大鼠分为高糖组和正常血糖组。每组又分假手术组,缺血再灌注组,右美托嘧啶(Dexmedetomide,Dex)预处理组。采用TUNEL进行凋亡指数检测以及Western blot法对Bax、Bcl-2蛋白表达、AKT磷酸化水平(p-AKT)进行检测。结果：急性高糖明显增加缺血再灌注后肾组织凋亡指数,Bax, AKT磷酸化水平,降低Bcl-2表达。正常血糖环境下,Dex能明显降低缺血再灌注所致的Bax升高水平,明显升高缺血再灌注所致的Bcl-2, p-AKT降低水平；而在高糖环境下,Dex的这种作用被明显减弱。结论：急性高糖加重缺血再灌注损伤肾组织的凋亡,急性高糖消除了Dex预处理对肾缺血再灌注损伤的保护作用,其机制可能与高糖环境下Dex对凋亡蛋白的调控消失,减弱了p-AKT水平的恢复有关。
[Abstract]:Acute hyperglycemia refers to the phenomenon that the blood glucose level is significantly increased due to some reasons. Current clinical studies, including kidney transplantation, have found that perioperative hyperglycemia is closely related to the prognosis of patients with renal ischemia-reperfusion. However, the specific impact on the mechanism is still unclear. In this study, a model of acute I / R hyperglycemia in kidney of normal SD rats was established to evaluate the effects of acute high glucose on renal I / R damage and to explore the possible mechanism of acute I / R injury. In addition, the effect of dextropyrimidine on the acute hyperglycemia model and its possible mechanism were discussed. Part I effects of acute hyperglycemia on renal ischemia-reperfusion injury in rats objective: to investigate the effects of acute hyperglycemia before ischemia on renal I / R in normal rats. Methods: the renal I / R acute hyperglycemic model of normal SD rats was established. The effects of acute hyperglycemia before ischemia on I / R in normal rats were studied by morphological and functional methods compared with those of simple kidney I / R. Results: acute hyperglycemia before ischemia increased renal ischemia-reperfusion injury, and Cr and BUN increased significantly 24 hours after reperfusion compared with non-hyperglycemic I / R group. Conclusion: acute hyperglycemia can aggravate renal I / R damage in normal rats. The second part is the immune inflammatory response of renal ischemia reperfusion injury in rats with acute hyperglycemia. Objective: to investigate the effect of acute high glucose concentration before ischemia on renal I / R immune inflammation in non-diabetic rats. Methods: the activity of MPO in renal homogenate was determined by using myeloperoxidase (MPO) assay kit by spectrophotometry. IL-1 尾, interferon- 纬 (IFN- 纬) and TLR4,TLR2, high mobility group protein B1 (high-mobility group box 1 HMGB1) in renal tissue were determined by immunohistochemistry, RT-PCR and RT-PCR,Western blot respectively. Expression of Cyclooxygenase 2 (Cox-2). Results: acute hyperglycemia increased the activity of MPO and the expression of IL-1 尾, IFN- 纬 and TLR4, TLR2,HMGB1,Cox-2 in renal tissue after ischemia reperfusion. Conclusion: acute hyperglycemia exacerbates the immune inflammatory injury induced by renal ischemia and reperfusion. The effect of acute high glucose on renal ischemia-reperfusion injury in rats and the effect of dextropyrimidine preconditioning objective: effects of acute high glucose on renal ischemia-reperfusion injury in rats and desmetropyrimidine preconditioning Methods: non-diabetic SD rats were divided into high glucose group and normal blood glucose group. Each group was divided into sham operation group, ischemia reperfusion group and Dexmedetomide,Dex preconditioning group. The expression of Bax,Bcl-2 protein and the level of AKT phosphorylation (p-AKT) were detected by TUNEL and Western blot. Results: acute hyperglycemia significantly increased the level of, Bax, AKT phosphorylation and decreased the expression of Bcl-2 in renal tissue after ischemia reperfusion. In normal blood glucose environment, Dex could significantly decrease the level of Bax and increase the level of Bcl-2, p-AKT induced by ischemia-reperfusion, but in high glucose environment, the effect of Dex was significantly weakened. Conclusion: acute hyperglycemia exacerbates the apoptosis of renal tissue after ischemia reperfusion injury, and the protective effect of Dex preconditioning on renal ischemia-reperfusion injury is eliminated. The mechanism may be related to the disappearance of Dex regulation on apoptotic protein in high glucose environment. Weakened the recovery of p-AKT levels related.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R699.2
[Abstract]:Acute hyperglycemia refers to the phenomenon that the blood glucose level is significantly increased due to some reasons. Current clinical studies, including kidney transplantation, have found that perioperative hyperglycemia is closely related to the prognosis of patients with renal ischemia-reperfusion. However, the specific impact on the mechanism is still unclear. In this study, a model of acute I / R hyperglycemia in kidney of normal SD rats was established to evaluate the effects of acute high glucose on renal I / R damage and to explore the possible mechanism of acute I / R injury. In addition, the effect of dextropyrimidine on the acute hyperglycemia model and its possible mechanism were discussed. Part I effects of acute hyperglycemia on renal ischemia-reperfusion injury in rats objective: to investigate the effects of acute hyperglycemia before ischemia on renal I / R in normal rats. Methods: the renal I / R acute hyperglycemic model of normal SD rats was established. The effects of acute hyperglycemia before ischemia on I / R in normal rats were studied by morphological and functional methods compared with those of simple kidney I / R. Results: acute hyperglycemia before ischemia increased renal ischemia-reperfusion injury, and Cr and BUN increased significantly 24 hours after reperfusion compared with non-hyperglycemic I / R group. Conclusion: acute hyperglycemia can aggravate renal I / R damage in normal rats. The second part is the immune inflammatory response of renal ischemia reperfusion injury in rats with acute hyperglycemia. Objective: to investigate the effect of acute high glucose concentration before ischemia on renal I / R immune inflammation in non-diabetic rats. Methods: the activity of MPO in renal homogenate was determined by using myeloperoxidase (MPO) assay kit by spectrophotometry. IL-1 尾, interferon- 纬 (IFN- 纬) and TLR4,TLR2, high mobility group protein B1 (high-mobility group box 1 HMGB1) in renal tissue were determined by immunohistochemistry, RT-PCR and RT-PCR,Western blot respectively. Expression of Cyclooxygenase 2 (Cox-2). Results: acute hyperglycemia increased the activity of MPO and the expression of IL-1 尾, IFN- 纬 and TLR4, TLR2,HMGB1,Cox-2 in renal tissue after ischemia reperfusion. Conclusion: acute hyperglycemia exacerbates the immune inflammatory injury induced by renal ischemia and reperfusion. The effect of acute high glucose on renal ischemia-reperfusion injury in rats and the effect of dextropyrimidine preconditioning objective: effects of acute high glucose on renal ischemia-reperfusion injury in rats and desmetropyrimidine preconditioning Methods: non-diabetic SD rats were divided into high glucose group and normal blood glucose group. Each group was divided into sham operation group, ischemia reperfusion group and Dexmedetomide,Dex preconditioning group. The expression of Bax,Bcl-2 protein and the level of AKT phosphorylation (p-AKT) were detected by TUNEL and Western blot. Results: acute hyperglycemia significantly increased the level of, Bax, AKT phosphorylation and decreased the expression of Bcl-2 in renal tissue after ischemia reperfusion. In normal blood glucose environment, Dex could significantly decrease the level of Bax and increase the level of Bcl-2, p-AKT induced by ischemia-reperfusion, but in high glucose environment, the effect of Dex was significantly weakened. Conclusion: acute hyperglycemia exacerbates the apoptosis of renal tissue after ischemia reperfusion injury, and the protective effect of Dex preconditioning on renal ischemia-reperfusion injury is eliminated. The mechanism may be related to the disappearance of Dex regulation on apoptotic protein in high glucose environment. Weakened the recovery of p-AKT levels related.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R699.2
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