牛化脓隐秘杆菌病肾脏Caspase-3、Caspase-8、Caspase-9、Bax和Bcl-2的表达
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摘要
通过研究牛化脓隐秘杆菌感染牛肾脏组织的病理学变化及检测Caspase-3、Caspase-8、Caspase-9、Bax和Bcl-2的表达,来确定牛肾脏组织的坏死和细胞凋亡情况。本研究首先筛检5头自然感染化脓隐秘杆菌病牛和2头阴性健康对照牛,采集其肾脏组织样本。采用H.E.染色法观察病理组织学变化;采用免疫组织化学方法来检测肾脏组织中与细胞正负凋亡有关的调节基因Bax和Bcl-2的表达情况以及凋亡通路蛋白Caspase-3、Caspase-8和Caspase-9的表达情况。结果显示,感染牛肾脏组织有坏死,间质有中性粒细胞浸润;感染组牛肾脏肾小球内皮细胞和肾小管上皮细胞Bax、Caspase-3和Caspase-9表达呈阳性,与对照组相比,差异显著;有少量肾小管上皮细胞、肾小球内皮细胞Caspase-8和Bcl-2表达呈阳性,但与对照组相比差异不显著。结果表明,化脓隐秘杆菌能够引起牛肾脏组织实质细胞发生坏死和凋亡,且以线粒体凋亡途径为主。
This experiment was conducted to determine whether Arcanobacterium pyogenes(A.pyogenes)could cause bovine kidneys necrosis and their parenchymal cells apoptosis by studying the pathological change and the expression of Caspase-3,Caspase-8,Caspase-9,Bax and Bcl-2 of the bovine kidneys infected with A.pyogenes.Specimens of bovine kidneys from 2 healthy dairy cow and 5 cattle infected with A.pyogenes were selected.Methods of H.E.and immunohistochemistry were exploited to research the histopathologic changes and investigate the expression of Caspase-3,Caspase-8,Caspase-9,Bax and Bcl-2 of the kidneys.Bax and Bcl-2 are two regulative genes related to cell apoptosis in positive and negative aspect,respectively.Caspase-3,Caspase-8 and Caspase-9 are associated proteins in apoptosis pathway.The results showed that the infective kidneys expressed necrosis,with their matrix infiltrated by neutrophils.Bax,Caspase-3 and Caspase-9 were expressed in the renal glomerular endothelial cells and renal tubular epithelial cells in the infected kidneys.There were few expression of Caspase-8 and Bcl-2 in the infected kidneys.The number of positive expressed cells of Bax,Caspase-3 and Caspase-9 was greater in the infected group than that in the control group(P<0.05).No significant difference of the expression of Caspase-8 and Bcl-2 was found between two groups(P >0.05).The results suggested that A.pyogenes could cause the parenchymal cells of the host kidneys necrosis,apoptosis and the mitochondria signaling pathways dominated in the pathogenesis of A.pyogenes.
This experiment was conducted to determine whether Arcanobacterium pyogenes(A.pyogenes)could cause bovine kidneys necrosis and their parenchymal cells apoptosis by studying the pathological change and the expression of Caspase-3,Caspase-8,Caspase-9,Bax and Bcl-2 of the bovine kidneys infected with A.pyogenes.Specimens of bovine kidneys from 2 healthy dairy cow and 5 cattle infected with A.pyogenes were selected.Methods of H.E.and immunohistochemistry were exploited to research the histopathologic changes and investigate the expression of Caspase-3,Caspase-8,Caspase-9,Bax and Bcl-2 of the kidneys.Bax and Bcl-2 are two regulative genes related to cell apoptosis in positive and negative aspect,respectively.Caspase-3,Caspase-8 and Caspase-9 are associated proteins in apoptosis pathway.The results showed that the infective kidneys expressed necrosis,with their matrix infiltrated by neutrophils.Bax,Caspase-3 and Caspase-9 were expressed in the renal glomerular endothelial cells and renal tubular epithelial cells in the infected kidneys.There were few expression of Caspase-8 and Bcl-2 in the infected kidneys.The number of positive expressed cells of Bax,Caspase-3 and Caspase-9 was greater in the infected group than that in the control group(P<0.05).No significant difference of the expression of Caspase-8 and Bcl-2 was found between two groups(P >0.05).The results suggested that A.pyogenes could cause the parenchymal cells of the host kidneys necrosis,apoptosis and the mitochondria signaling pathways dominated in the pathogenesis of A.pyogenes.
引文
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[2]范春玲,周玉龙,任亚超,等.动物组织石蜡切片的绿色环保制作技术.CN201110175687.9[P].2013.09.
[2]SEMAMBO D K,AYLIFFE T R,BOYD J S,et al.Early abortion in cattle induced by experimental intrauterine infection with pure cultures of Actinomyces pyogenes[J].Vet Rec,1991,129:12-16.
[3]鲁杏华,何世成,谈志祥,等.猪化脓隐秘杆菌的分离与鉴定[J].中国畜牧兽医,2009,36(5):149-151.
[4]张素辉,付利芝,黄勇富,等.山羊化脓隐秘杆菌的分离鉴定与分析[J].中国兽医杂志,2015,51(6):37-39.
[5]刘明春,刘耀川,赵敬翠,等.奶牛子宫内膜炎化脓隐秘杆菌16SrRNA基因的鉴定与分析[J].中国兽医学报,2009,29(3):343-345.
[6]郭婷,齐欣,赵秀宇,等.金黄色葡萄球菌引起猪DIC病理学观察[J].中国人兽共患病学报,2009,25(12):1222-1223.
[7]王密,周玉龙,朴范泽.牛化脓隐秘杆菌病PCR诊断方法的建立[J].微生物学通报,2010,37(9):1320-1324.
[8]周玉龙,高小兵,范春玲,等.牛源化脓隐秘杆菌PLO蛋白生物学功能的初步研究[J].中国预防兽医学报,2012,6(3):45-48.
[9]郭婷,齐欣,张丽,等.牛隐秘杆菌病的病理学研究[J].黑龙江畜牧兽医,2010(1):127-128.
[10]LALIER L,CARTRON P F,JUIN P,et al.Bax activation and mitochondrial insertion during apoptosis[J].Apoptosis,2007,12(5):887-896.
[11]MATSUSHITA K,WU Y,QIU J,et al.Fas receptor and neuronal cell death after spinal cord ischemia[J].J Neurosci,2000,20(18):6879-6887.
[12]LAI G H,ZHAND Z,SIRICA A E.Celecoxibacts in a cyclooxy-genase-2-independent manner and in synergy with emodin to suppress rat cholangiocarcinoma growth in vitro through a mechanism involving enhanced Akt inactivation and increased activation of caspases-9and-3[J].Mol Cancer Ther,2003,2(3):265-271.