摘要
目的:采用低氧培养HKC细胞株,检测HKC STAT 1、α-SMA以及HIF-1α的动态表达,探讨JAK/STAT信号转导途径在低氧诱导的肾小管上皮上皮-间质转化中的作用。方法:将对数生长期HKC随机分为常规培养组(A组),常规+Flu培养组(B组),低氧培养组(C组),低氧+Flu培养组(D组)4组。C组和D组采用三气培养箱低氧培养。分别在实验第1、3、6d观察细胞形态改变情况,免疫细胞化学检测HIF-1α、α-SMA以及STAT1表达。结果 :与A、B组比较,C、D组细胞生长缓慢,随着低氧培养时间的延长,细胞形态变细长,呈梭形。HIF-1α在A、B组表达极低,C、D组表达显著增强(P<0.05),各时间点差异无统计学意义。从第3 d开始C组STAT1、α-SMA表达显著高于A组(P<0.05)。与C组相比,D组各时间点STAT1表达显著降低(P<0.05),α-SMA从第3 d开始表达明显降低(P<0.05)。结论:低氧可能通过JAK/STAT1信号通路诱导肾小管上皮细胞发生表型转化,从而促进肾间质纤维化的发展。
Objective： To observe the expression of HIF-1α、α-SMA, and STAT1 in the human kidney proximal epithelial cell（HKC） under the condition of hypoxia and to investigate the effect of JAK/STAT signaling pathway on EMT induced by hypoxia. Methods： Use a tri-gas incubator to build the model of EMT induced by hypoxia. The cells in exponential phase were divided into four groups randomly. They are routine culture（group A）, routine culture ＋ Flu（group B）, hypoxia culture（group C）, and hypoxia culture＋Flu（group D）. The morphology of cells weas observed under microscope and the expression of α-SMA, STAT1 and HIF-1α was evaluated by immunohistochemistry on day 1, 3, and 6. Results： Cells grow slower in groups C and D compared to groups A and B under hypoxia. As time goes by, cells under hypoxia become heterogeneous in morphology, showing spindle-shaped. The expression of HIF-1α is increased in group C and group D compared to cells cultured in routine incubator in groups A and B, which hardly express HIF-1 α. Compared with the group A, cells in group C express much more STAT1 and α-SMA（P〈0.05） on day 3 and 6. Compared with the group C, cells in group D express lower levels of STAT1（P〈0.05） and α-SMA（P〈0.05） at each time point on day 3 and 6.Conclusion： The JAK/STAT1 signaling pathway may participate in the process of EMT of HKC induced by hypoxia and thus play an important role in the process of renal interstitial fibrosis.
出处
《泸州医学院学报》
2016年第1期34-37,共4页
Journal of Luzhou Medical College
基金
四川省科技厅重点项目(2010JY0080)]
