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目的:探讨renalase在人近曲肾小管上皮细胞系(HK-2)的表达与分泌,为进一步研究细胞水平renalase及其通路建立稳定的实验平台。方法:以HK-2细胞系作为研究材料。①应用Westernblot方法检测renalase蛋白的表达。②用real-timePCR方法检测renalasemRNA表达的变化。③用ELISA方法检测细胞上清液中renalase的浓度。结果:在mRNA水平及蛋白水平均检测到renalase表达。结论:首次在mRNA水平及蛋白水平证实了HK-2细胞能够表达renalase,为进一步研究儿茶酚胺或缺血缺氧刺激下细胞renalase的表达奠定了基础。
OBJECTIVE: To investigate the expression and secretion of renalase in human proximal tubular epithelial cell line (HK-2), and to establish a stable experimental platform for further study on renalase and its pathway at cellular level. Methods: HK-2 cell line as a research material. ① Western blot was used to detect the expression of renalase protein. ② using real-timePCR detection of renalase mRNA expression changes. ③ ELISA method for the detection of cell supernatant renalase concentration. Results: Renalase expression was detected both at mRNA and protein levels. CONCLUSION: HK-2 cells can express renalase at the mRNA and protein levels for the first time, which lays the foundation for further study on the expression of renalase induced by catecholamine or ischemia-hypoxia.