摘要
目的探讨自噬介导铁死亡在苯诱导的大鼠血液毒性中的作用。方法将80只Wistar雄性大鼠随机分为4组,对照组、染苯组(BZ)、染苯+铁死亡抑制剂组(BZ+Fer-1)和染苯+自噬抑制剂组(BZ+3MA),每组20只。BZ组按照注射体积200μl/100 g于大鼠皮下注射苯油混合物,染毒浓度为800 mg/kg;对照组注射等量玉米油;抑制剂组按照400μl/100 g给药量进行大鼠腹腔注射,抑制剂浓度为10 mg/kg,每周注射3次,连续给药4周。分别于第7、14、21、28天观察大鼠体质量变化;眼内眦静脉取血检测各组大鼠血常规。以异氟烷麻醉大鼠脱臼处死后心脏取血并分离血清,检测大鼠血清中丙二醛(MDA)和亚铁离子(Fe^(2+))水平变化;分离一侧股骨,采用HE染色法观察各组大鼠股骨病理组织学变化,收集股骨骨髓单个核细胞(BMMNCs);蛋白免疫印迹(Western blot)法检测股骨骨髓细胞转铁蛋白受体(TFRC)、长链脂酰辅酶A合成酶4(ACSL4)、谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链(FTH1)和自噬相关蛋白(p62、LC3Ⅱ/Ⅰ)等相关蛋白的表达水平。结果与对照组相比,BZ组21 d后大鼠体质量显著下降(P<0.05);14 d后外周血WBC、21 d后RBC和Hb显著下降(P<0.05)。股骨病理检测结果显示,BZ组大鼠股骨骨髓造血细胞减少,脂肪细胞增多;血清MDA、Fe^(2+)水平及BMMNCs中TFRC、ACSL4、LC3Ⅱ/Ⅰ蛋白表达水平较对照组显著增高(P<0.05),GPX4、p62、FTH1蛋白表达水平显著降低(P<0.05)。与BZ组大鼠相比,BZ+Fer-1组和BZ+3MA组大鼠体质量和血WBC计数显著增加,外周血RBC和Hb水平明显增加,且显著高于对照组(P<0.05);MDA、Fe^(2+)含量以及TFRC、ACSL4、LC3Ⅱ/Ⅰ蛋白表达水平显著降低(P<0.05),GPX4、FTH1、p62蛋白表达水平显著增高(P<0.05)。结论苯可诱导大鼠造血机能障碍,自噬介导的铁死亡在苯诱导大鼠血液毒性发生中具有重要作用。
Objective To explore the role of autophagy mediated ferroptosis in benzene induced hematotoxicity.Methods Eighty Wistar male rats were randomly divided into four groups:control group(Control),benzene exposed group(BZ),ben⁃zene exposed+ferroptosis inhibitor group(BZ+Fer⁃1)and benzene exposed+autophagy inhibitor group(BZ+3MA)with 20 rats in each group.The exposure concentration of benzene was 800 mg/kg,subcutaneously;the control rat was injected with the same amount of corn oil;the rats of two inhibitor groups were intraperitoneally injected with concentration of 10 mg/kg,all rats were received injection three times a week,for four consecutive weeks.The body⁃weight of rats were observed at the 7 th,14 th,21st and 28th day after injection respectively,and blood sample was taken from the vein of inner canthus,for blood routine exam⁃ination,then the rats were anesthetized with isoflurane for blood⁃taking from heart and the serum was separated for the detection of malondialdehyde(MDA)and Fe^(2+),and one femur was separated for the pathological observation and the bone marrow mono⁃nuclear cells(BMMNCs)were collected for the detection of the expression levels of ferroptosis related proteins such as transferrin receptor(TFRC),long⁃chain acyl coenzyme A synthetase 4 hileo(ACSL4),glutathione peroxidase 4(GPX4),ferritin heavy chain(FTH1)and autophagy related proteins(p62,LC3Ⅱ/Ⅰ)by Westren blot method.Results The results showed that compared with control group,the body⁃weight of benzene exposed rats were decreased significantly after 21 days of exposure(P<0.05),the number of WBC in peripheral blood were significantly decreased after 14 days exposure(P<0.05),and the amounts of RBC and Hb in peripheral blood were also decreased significantly after 21 days of exposure(P<0.05);the pathological exam⁃ination showed that the hematopoietic cells in bone marrow decreased,while the number of adipocytes were increased;furthermore,the levels of MDA and Fe^(2+)in serum were significantly higher(P<0.05),the expression
作者
董双燕
杨晓涵
张晓霞
厉铭
李超
孟祥敬
王真真
贾强
DONG Shuangyan;YANG Xiaohan;ZHANG Xiaoxia;LI Ming;LI Chao;MENG Xiangjing;WANG Zhenzhen;JIA Qiang(Shandong Provincial Academy of Occupational Health and Occupational Medicine,Shandong First Medical University/Shandong Provincial Academy of Medical Science,Jinan,Shandong 250062,China)
出处
《中国工业医学杂志》
CAS
2023年第2期111-115,F0003,共6页
Chinese Journal of Industrial Medicine
基金
济南市科技发展计划项目(201907004)
山东第一医科大学学术提升计划(2019QL001)。
关键词
苯
铁死亡
自噬
血液毒性
benzene
ferroptosis
autophagy
hematotoxicity研究发现[1],
