他克莫司预处理对肝脏缺血再灌注损伤的作用

发布时间：2018-03-16 00:22

本文选题：他克莫司　切入点：缺血再灌注　出处：《天津医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨他克莫司预处理对大鼠肝脏缺血再灌注损伤的作用及其与肝脏缺血再灌注期HMGB1、HIF-1α和HO-1的关系。方法:将64只成熟SD大鼠随机分为2大组,每组32只,分别应用于建立大鼠肝脏热IR模型和冷IR模型中。每种IR模型各分成4个小组,分别为假手术组、缺血再灌注组(对照组)、FK506低剂量处理组、FK506高剂量处理组。术后再灌注6h取材,比较各组大鼠血清转氨酶水平;利用ELISA法检测血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)水平;采用HE染色观察肝组织病理改变;通过实时荧光定量PCR(RT-q PCR)来观察FK506预处理对HMGB1、血红素加氧酶-1(heme oxygenase-1,HO-1)m RNA表达的影响;免疫组织化学法检测HMGB1的分布变化;Western blot法检测HMGB1、HIF-1α和HO-1蛋白表达情况。结果:经FK506处理后的给药组中,两种IR模型大鼠转氨酶ALT、AST,炎症因子TNF-α、IL-6和IL-1β含量较未处理对照组均有不同程度降低。两种IR模型中的对照组内均观察到肝血窦淤血、中性粒细胞浸润和片状坏死。相比之下,不同剂量的FK506处理组中病理学改变均明显减轻。与假手术组相比,行IR的对照组中HMGB1的m RNA及蛋白表达显著增加,但经FK506预处理后表达明显减少。在经低剂量FK506预处理的大鼠中,热IR模型和冷IR模型组的HIF-1α、HO-1表达均呈现明显增加。结论:FK506预处理可以显著减轻大鼠肝脏IR损伤,该作用可能是FK506通过降低HMGB1的表达,抑制炎症因子释放及上调HIF-1α、HO-1表达发挥的。
[Abstract]:Objective: to investigate the effect of tacrolimus preconditioning on hepatic ischemia-reperfusion injury in rats and its relationship with HMGB1HIF-1 伪 and HO-1 during hepatic ischemia-reperfusion. Methods: 64 adult SD rats were randomly divided into two groups, 32 rats in each group. Each IR model was divided into four groups: sham-operated group and ischemia-reperfusion group (control group, FK506 low-dose treatment group, FK506 high-dose treatment group). The levels of serum aminotransferase (alt), tumor necrosis factor- 伪 (TNF- 伪), interleukin-6 (IL-6) and interleukin-1 尾 interleukin-1 尾 (IL-1 尾) were detected by ELISA assay, and the pathological changes of liver tissue were observed by HE staining. The effect of FK506 pretreatment on the expression of HMGB1, hemhemoxygenase-1 (HO-1) RNA was observed by real-time fluorescence quantitative PCR(RT-q. Immunohistochemical method was used to detect the distribution of HMGB1. The expression of HMGB1HIF-1 伪 and HO-1 protein was detected by Western blot. Results: in the group treated with FK506, the expression of HIF-1 伪 and HIF-1 伪 was detected by Western blot. The levels of alt, TNF- 伪, IL-6 and IL-1 尾 in the two IR models were significantly lower than those in the untreated control group. The hepatic sinusoidal congestion, neutrophil infiltration and flake necrosis were observed in the two IR models. Compared with sham-operated group, the expression of m RNA and protein of HMGB1 in the control group treated with IR was significantly increased. However, after pretreatment with FK506, the expression of HIF-1 伪 -tHO-1 was significantly increased in the rats pretreated with low-dose FK506. Conclusion the expression of HIF-1 伪 -HO-1 in the hot IR model group and the cold IR model group was significantly increased. Conclusion: FK506 pretreatment can significantly attenuate the hepatic IR injury in rats. This effect may be played by FK506 by reducing the expression of HMGB1, inhibiting the release of inflammatory factors and upregulating the expression of HIF-1 伪 -HO-1.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R657.3

【参考文献】