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中国临床研究英文版:2021,34(9):1159-1163
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p53/SAT1/ALOX15铁死亡通路蛋白与特发性炎性肌病的相关性
(1. 青海大学,青海 西宁 810000;2. 青海大学附属医院风湿免疫科,青海 西宁 810000)
Correlation between p53/SAT1/ALOX15 ferroptosis pathway and idiopathic inflammatory myopathy
摘要
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Received:April 22, 2021   Published Online:September 20, 2021
中文摘要: 目的 通过免疫组化法分别检测实验性自身免疫性肌炎(EAM)小鼠模型肌肉组织中p53、精脒/精胺N1-乙酰基转移酶1(SAT1)、花生四烯酸-15-脂加氧酶(ALOX15)蛋白的表达,探讨特发性炎性肌病(IIM)的发病机制与p53/SAT1/ALOX15铁死亡通路蛋白的相关性。方法 随机将13只BALB/c小鼠分为EAM模型组(n=7)、对照组(n=6),通过粗略提取的豚鼠骨骼肌匀浆与完全弗氏佐剂混匀后皮下注射建立EAM小鼠模型,在末次免疫处理后1周,通过对小鼠临床表现的观察和体重、四肢的肌力、血清肌酸激酶水平、骨骼肌HE染色结果的测定等综合方法评定是否造模成功。通过免疫组化法分别检测EAM小鼠及对照组小鼠肌肉组织中p53、SAT1、ALOX15蛋白表达水平。 结果 经综合方法判定,EAM组小鼠6只造模成功(1只死亡)。EAM模型组小鼠肌肉组织中p53、SAT1、ALOX15三种蛋白的阳性表达率及免疫组化表达评分均高于对照组(P<0.05,P<0.01)。Pearson相关分析结果显示,p53分别与SAT1、ALOX15呈正相关(r=0.976,P=0.001;r=0.963,P=0.002),SAT1与ALOX15呈正相关(r=0.951,P=0.004)。结论 p53/SAT1/ALOX15铁死亡通路蛋白可能参与了特发性炎性肌病的发病。
Abstract:Objective By detecting the expression of p53, spermidine/spermine N1-acetyltransferase 1 (SAT1) and arachidonate 15-lipoxygenase (ALOX15) proteins in the muscle tissue of experimental autoimmune myositis (EAM) mice with immunohistochemistry to explore the correlation between the pathogenesis of idiopathic inflammatory myopathy (IIM) and p53/SAT1/ALOX15 ferroptosis pathway protein. Methods Thirteen BALB/c mice were randomly divided into EAM group (n=7) and control group (n=6). EAM mouse model was established by subcutaneous injection after roughly extracted guinea pig skeletal muscle homogenate was mixed with complete Freund′s adjuvant. One week after the last immune treatment, observing the clinical manifestations of mice and measuring the weight, muscle strength of limbs, serum CK level and hematoxylineosin staining results of skeletal muscle were used to determine whether the model was successful. The expression of p53, SAT1 and ALOX15 proteins in mice muscle tissues were detected by immunohistochemistry. Results By the integrated method 6 mice in EAM group were successfully established (1 died). The positive expression rate and immunohistochemical expression score of p53, SAT1 and ALOX15 in muscle tissue of EAM model group were higher than those of control group (P<0.05, P<0.01). Pearson correlation analysis showed that p53 was positively correlated with SAT1 and ALOX15 respectively (r=0.976, P=0.001; r=0.963, P=0.002), SAT1 was positively correlated with ALOX15 (r=0.951, P=0.004). Conclusion p53/SAT1/ALOX15 ferroptosis pathway protein may be involved in the pathogenesis of idiopathic inflammatory myopathy.
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基金项目:青海省科技厅应用基础研究科技计划项目(2019-ZJ-7094)
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