大鼠肢体缺血再灌注急性肺损伤细胞自噬及其相关蛋白的表达

发布时间：2018-02-16 09:59

  本文关键词： 呼吸窘迫综合征 成人 自噬 肢体缺血再灌注 Beclin LC　出处：《重庆医学》2014年32期 　论文类型：期刊论文

【摘要】：目的研究大鼠双下肢缺血再灌注所致急性肺损伤(LIR-ALI)肺组织细胞早期自噬及其相关蛋白的表达。方法采用LIR-ALI模型,12只雄性SD大鼠,以随机数字表法分为假手术(Sham)组,肢体缺血再灌注(I/R)组,每组6只。采用ELISA检测血清白乳酸脱氢酶(LDH)和免疫荧光法检测肺组织病理变化,利用RT-PCR检测各组肺组织Beclin1和Atg5mRNA表达及Western blot检测LC3蛋白的表达。结果与Sham组比较,I/R组LDH值升高,显示大鼠LIR-ALI模型造模成功且肺组织免疫荧光特异性标记抗体高表达(P0.01)。采用2-△△CT法分析肺组织Beclin1和Atg5mRNA相对表达量,在I/R组较Sham组升高(P0.01);Western blot法检测显示,I/R组较Sham组LC3-Ⅰ、LC3-Ⅱ高表达、LC3-Ⅱ/GAPDH亦明显升高(P0.01)。结论大鼠LIR-ALI后激活肺组织细胞自噬及其相关蛋白高表达。
[Abstract]:Objective to study the expression of early autophagy and its associated proteins in lung tissue of rats with acute lung injury induced by ischemia-reperfusion of lower extremities. Methods Twelve male SD rats with LIR-ALI model were randomly divided into sham operation group. ELISA and immunofluorescence were used to detect the pathological changes of lung tissue in the I / R group, 6 rats in each group, and 6 rats in each group. The serum white lactate dehydrogenase (LDH) and immunofluorescence assay were used to detect the pathological changes of lung tissue. The expression of Beclin1 and Atg5mRNA in lung tissue of each group was detected by RT-PCR and the expression of LC3 protein was detected by Western blot. Results the LDH value of I / R group was higher than that of Sham group. The results showed that the rat model of LIR-ALI was successful and the lung tissue immunofluorescence specific antibody was highly expressed P0.01. The relative expression of Beclin1 and Atg5mRNA in lung tissue was analyzed by 2-CT method. Compared with Sham group, the I / R group was significantly higher than that in Sham group by Western blot assay. The results showed that the higher expression of LC3- 鈪，

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