摘要
本研究旨在探讨低氧诱导因子-1α(HIF-1α)对牦牛肾小管上皮细胞上皮-间质转化(EMT)的影响。首先对牦牛肾小管上皮细胞进行培养鉴定,并选取第三代细胞对其分别进行药物处理(DMOG组)和不做药物处理(对照组),检测EMT相关基因在mRNA和蛋白水平的表达变化,并观察细胞形态变化。结果表明:使用Ⅰ+Ⅱ型胶原酶可以更好地获得肾小管上皮细胞;免疫荧光结果细胞CK18表达呈阳性,Vimentin和CD31呈阴性表达;qRT-PCR和Western blot结果表明,DMOG组中的HIF-1α,TGF-β1,α-SMA在mRNA和蛋白水平均较对照组有极显著增加(P<0.01),而E-cadherin的表达显著降低(P<0.05);经药物干预后,细胞形态由铺路石样转变为长梭形,经鉴定长梭形细胞为间质细胞。本研究成功建立了牦牛肾小管上皮细胞的分离培养体系,并发现HIF-1α表达上调可以促进牦牛肾小管上皮细胞上皮-间质转化。
The aim of this study was to discuss the effect of hypoxia-inducible factor 1α(HIF-1α) on renal tubular epithelial-to-mesenchymal transition(EMT) in yak. First of all, the primary culture and identification of yak renal tubular epithelial cells were conducted. Then the third generation cells were chosen to be treated with drugs(DMOG group) and untreated(control group). The expression changes of EMT related genes were detected at both mRNA and protein levels, and the morphological changes of cells were observed. The results showed that renal epithelial cells could be better obtained by using the collagenase Ⅰ+Ⅱ;Immunofluorescence results showed that CK18 expression was positive, but Vimentin and CD31 were negative;The results of qRT-PCR and Western blot showed that mRNA and protein expression levels of HIF-1α, TGF-β1 and α-SMA in DMOG group were extremely significantly higher than those in control group(P<0.01), while the expression of E-cadherin was significantly lower than that in control group(P<0.05). After drug intervention, the cell morphology changed from paving-stone shape to long shuttle, which was identified as mesenchymal cells. Therefore, this study had successfully established a method for isolation and culture of yak renal tubular epithelial cells, and proved that the up-regulation of HIF-1α expression could promote the renal tubular epithelial-to-mesenchymal transition of yak.
作者
白雪峰
崔燕
余四九
李慧
何俊峰
廖博
BAI Xuefeng;CUI Yan;YU Sijiu;LI Hui;HE Junfeng;LIAO Bo(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep&Goat,Lanzhou 730070,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2020年第3期620-629,共10页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31772691)。
关键词
牦牛
肾小管上皮细胞
分离培养
HIF-1Α
上皮-间质转化
yak
real tubular epithelial cell
isolation and culture
HIF-1α
epithelial-to-mesenchymal transition
