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阿维菌素致鸽子大脑组织程序性坏死研究

Study on necroptosis of pigeon brain tissue induced by avermectin exposure

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【作者】 闫立地邱思元王雨欣刘畅翟祖欢杨攀曲丽娜张奕婷

【Author】 YAN Lidi;QIU Siyuan;WANG Yuxin;LIU Chang;ZHAI Zuhuan;YANG Pan;QU Li’na;ZHANG Yiting;School of Biological Engineering, Daqing Normal University;School of Basic Medicine, Guizhou Medical University;

【机构】 大庆师范学院生物工程学院贵州医科大学基础医学院

【摘要】 为了研究阿维菌素(AVM)对白羽王鸽大脑组织中细胞程序性坏死的影响,试验将50只3~4月龄白羽王鸽随机分成5组,分别为空白组(饲喂基础日粮)、30L组(日粮中添加20 mg/kg AVM,饲喂30 d)、30H组(日粮中添加60 mg/kg AVM,饲喂30 d)、60L组(日粮中添加20 mg/kg AVM,饲喂60 d)、60H组(日粮中添加60 mg/kg AVM,饲喂60 d),每组10只,剖杀取大脑组织后通过光学显微镜观察脑组织细胞形态并检测超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量,通过实时荧光定量PCR(qPCR)和Western-bolt方法分别检测程序性坏死相关基因肿瘤坏死因子-α(TNF-α)、受体相互作用蛋白激酶1(RIPK-1)、受体相互作用蛋白激酶3(RIPK-3)和混合系列蛋白激酶结构域样蛋白(MLKL)的mRNA和蛋白质表达情况。结果表明:与空白组比较,30H组SOD、CAT活性显著降低(P<0.05);60L组SOD活性显著降低(P<0.05),CAT活性极显著降低(P<0.01),MDA含量显著升高(P<0.05);60H组SOD、CAT活性极显著降低(P<0.01),MDA含量极显著升高(P<0.01)。与空白组比较,60L组TNF-α、RIPK-1、MLKL的mRNA表达量显著升高(P<0.05),RIPK-3的mRNA表达量极显著升高(P<0.01);60H组TNF-α、RIPK-3、MLKL的mRNA表达量极显著升高(P<0.01),RIPK-1的mRNA表达量显著升高(P<0.05)。与空白组比较,60L组RIPK-1、RIPK-3、MLKL蛋白表达量显著升高(P<0.05),TNF-α蛋白表达量极显著升高(P<0.01);60H组TNF-α、RIPK-1、RIPK-3蛋白表达量极显著升高(P<0.01),MLKL蛋白表达量显著升高(P<0.05)。说明AVM可导致白羽王鸽脑组织发生程序性坏死。

【Abstract】 In order to investigate the effects of avermectin(AVM) exposure on necroptosis of brain tissue of White King pigeon, in this experiment, the 50 3-to 4-month-old White King pigeons were randomly divided into 5 groups, namely blank group(feeding basal diet), 30L group(20 mg/kg AVM added to diet, feeding 30 d), 30H group(60 mg/kg AVM added to diet, feeding 30 d), 60L group(20 mg/kg AVM added to diet, feeding 60 d), 60H group(60 mg/kg AVM added to diet, feeding 60 d); 10 animals in each group. After dissection, the brain tissue was taken and the morphology of brain tissue cells was observed by light microscopy, and the activity of superoxide dismutase(SOD), catalase(CAT) and malondialdehyde(MDA) contents were detected. The mRNA and protein expressions of necroptosis-related genes including tumor necrosis factor-α(TNF-α), receptor-interacting protein kinase 1(RIPK-1), receptor-interacting protein kinase 3(RIPK-3), and mixed series protein kinase domain-like protein(MLKL) were detected by fluorescent quantitative PCR(qPCR) and Western-bolt methods, respectively. The results showed that compared with the blank group, the SOD and CAT activities in the 30H group were significantly reduced(P<0.05); SOD activity in the 60L group decreased significantly(P<0.05), CAT activity was significantly reduced(P<0.01), and MDA content was significantly increased(P<0.05); the activities of SOD and CAT in the 60H group was significantly reduced(P<0.01), and the content of MDA was significantly increased(P<0.01). Compared with the blank group, the mRNA expressions of TNF-α, RIPK-1 and MLKL in the 60L group increased significantly(P<0.05), and the mRNA expression of RIPK-3 increased significantly(P<0.01); the mRNA expressions of TNF-α, RIPK-3 and MLKL in the 60H group was significantly increased(P<0.01), and the expression of RIPK-1 mRNA was significantly increased(P<0.05). Compared with the blank group, the expressions of RIPK-1, RIPK-3 and MLKL proteins in the 60L group increased significantly(P<0.05), and the expression of TNF-α protein increased significantly(P<0.01); the expressions of TNF-α, RIPK-1 and RIPK-3 proteins in the 60H group was significantly increased(P<0.01), and the expression of MLKL protein was significantly increased(P<0.05). The results suggested that AVM could cause the necroptosis of brain tissue of White King pigeon.

【基金】 黑龙江省自然科学基金联合引导项目(LH2019C071)
  • 【文献出处】 黑龙江畜牧兽医 ,Heilongjiang Animal Science and Veterinary Medicine , 编辑部邮箱 ,2023年15期
  • 【分类号】S858.39
  • 【下载频次】13
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