促红细胞生成素对牦牛肾间质成纤维细胞凋亡因子表达的影响  

Effect of Erythropoietin on the Expression of Apoptotic Factor in Yak Renal Interstitial Fibroblasts

作　　者：卢增华 崔燕[1,2] 余四九 白雪峰[1] 卢鸿琴 何俊峰 卢凯 翟国亮[1] 祁正满 LU Zenghua;CUI Yan;YU Sijiu;BAI Xuefeng;LU Hongqin;HE Junfeng;LU Kai;ZHAI Guoliang;QI Zhengman(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Gansu Province Livestock Embryo Engineering Research Center,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070 [2]甘肃省牛羊胚胎工程技术研究中心,兰州730070

出　　处：《畜牧兽医学报》2024年第8期3460-3471,共12页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(32372974)。

摘　　要：旨在探讨促红细胞生成素(EPO)对牦牛肾间质成纤维细胞(RIFs)中凋亡因子的影响。本研究首先分别采集3头4岁龄的健康牦牛和黄牛的肾脏组织,采用免疫组织化学方法和Western blot检测牦牛和黄牛肾脏中EPO的准确分布及其表达量。同时对牦牛RIFs进行原代培养鉴定,并对其进行不同药物处理来调控RIFs中EPO表达水平,检测EPO对凋亡相关基因Bax,Bcl-2,PCNA在mRNA和蛋白表达水平的影响。免疫组化学检测结果显示,EPO在牦牛和黄牛肾脏中集合管上皮细胞、肾小管上皮细胞和小管周间质成纤维样细胞中均呈阳性表达。qRT-PCR和Western blot结果显示,EPO在牦牛肾脏中的表达量显著高于黄牛(P<0.001)。随后对生成EPO的RIFs进行原代培养并调控EPO的表达,结果表明:激活组显著升高RIFs中Bcl-2和PCNA的表达,降低Bax的表达(P<0.001)。EPO抑制组显著降低RIFs中Bcl-2、PCNA的表达,上调Bax的表达(P<0.001)。综上表明,EPO在牦牛和黄牛肾脏中集合管上皮细胞、肾小管上皮细胞和小管周间质成纤维样细胞中均呈阳性表达,牦牛肾脏中的EPO高表达能够促进RIFs中Bcl-2和PCNA的表达,并降低Bax的表达,说明EPO可以在牦牛肾脏中起到抗凋亡的作用。The purpose of this study was to investigate the effect of erythropoietin(EPO)on apoptosis factors in yak renal interstitial fibroblasts(RIFs).Firstly,the kidney tissues of three 3 healthy yaks and cattle aged 4 years were collected respectively,immunohistochemistry and Western blot were used to detect the exact distribution of EPO and its expression in the kidneys of yaks and calves.Meanwhile,the yak RIFs were identified by primary culture,and different drug treatments were applied to regulate the EPO expression level in the RIFs,and the effects of EPO on the apoptosis-related genes Bax,Bcl-2,and PCNA at mRNA and protein expression levels were detected.Immunohistochemical results showed that EPO was positively expressed in collecting duct epithelial cells,renal tubular epithelial cells,and peritubular mesenchymal fibroblast-like cells in the kidneys of both yaks and calves.qRT-PCR and Western blot results showed that the expression of EPO in the kidneys of yaks was significantly higher than that in the calves(P<0.001).Subsequently,the EPO-generating RIFs were cultured in primary culture and the expression of EPO was regulated,and the results showed that the activation group significantly elevated the expression of Bcl-2 and PCNA and decreased the expression of Bax in the RIFs(P<0.001).The EPO-inhibition group significantly decreased the expression of Bcl-2,PCNA and up-regulated the expression of Bax in the RIFs(P<0.001).In conclusion,it was showed that EPO was positively expressed in collecting duct epithelial cells,renal tubular epithelial cells and peritubular mesenchymal fibroblast-like cells in the kidney of yak and calf,and the high expression of EPO in the kidney of yak was able to promote the expression of Bcl-2 and PCNA and decrease the expression of Bax in the RIFs,which indicated that EPO could play an anti-apoptotic role in the kidney of yak.

关 键 词：牦牛 促红细胞生成素(EPO) 肾间质成纤维细胞(RIFs) 凋亡 

分 类 号：S823.85[农业科学—畜牧学]