摘要
目的探讨维生素D受体(vitamin D receptor,VDR)和二羧酸转运蛋白1(sodium-dicarboxylate cotransporter protein 1,Na DC1)在人肾近曲小管上皮细胞(HK-2 cells,HK-2细胞)中的表达及其对低枸橼酸尿症的意义.方法按照HK-2细胞培养的条件培养细胞,采用流式细胞学检测所培养的HK-2细胞的细胞凋亡,免疫细胞化学SP法检测VDR和Na DC1在HK-2细胞中的表达.结果细胞培养:拍照记录细胞状态;流式细胞学:1号至4号流式管HK-2细胞凋亡率分别为6.87%,6.80%,8.31%,4.274%;免疫细胞化学:Na DC1在HK-2细胞中表达,胞膜染色.Na DC1的阴性对照,胞膜未染色.Na DC1的阳性对照,在人胚肾293细胞(human embryonic kidney 293 cells,HEK293 cells)胞膜表达,胞膜染色.VDR在HK-2细胞中表达,核膜染色.VDR的阴性对照,核膜未染色.VDR的阳性对照,在人肺癌细胞系A549核膜表达,核膜染色.结论所培养的HK-2细胞的细胞凋亡率比较低,细胞状态较好.VDR和Na DC1分别在HK-2细胞核膜和胞膜表达.VDR和Na DC1都与低枸橼酸尿症的发生发展有关,推测VDR可能参与调控Na DC1活性或表达.
Objective To investigate expression of vitamin D receptor（VDR） and sodium-dicarboxylate cotransporter protein 1（Na DC1） in human renal proximal tubule epithelial cell（HK-2 cells） and their significance in hypocitraturia. Methods We cultured HK-2 cells under the cultivation condition of HK-2 cells. The apoptosis of HK-2 cells cultured was detected by Flow Cytometry,and the expression of VDR and Na DC1 in HK-2cells was detected by immunocytochemical SP method. Results The culture of HK-2 cells： The state of HK-2 cells was recorded by taking pictures. Flow cytometry： HK-2 cells apoptosis rate of flow tube number 1-4 were respectively6.87%,6.80%,8.31% and 4.274%. Immunocytochemistry ： The expression of Na DC1 in HK-2 cells： the cell membrane of HK-2 cells and Na DC1 positive control（human embryonic kidney 293 cells,HEK293 cells） were stained. While cell membrane of Na DC1 negative control was not stained. The expression of VDR in HK-2 cells： the nuclear membrane of HK-2 cells and VDR positive control（human lung cancer cell line A549 cells） were stained.While nuclear membrane of VDR negative control was not stained. Conclusions The apoptosis rate of the HK-2 cells cultured is relatively low,and the cells are in good condition. VDR and Na DC1 are respectively expressed in the nuclear membrane and cell membrane of HK-2 cells. Both VDR and Na DC1 are associated with the occurrence and development of hypocitraturia. We conclude that VDR may be involved in regulating the activity or expression of Na DC1.
出处
《昆明医科大学学报》
CAS
2016年第1期35-39,共5页
Journal of Kunming Medical University
基金
国家自然科学基金资助项目(81460141)
云南省科技厅-昆明医科大学联合专项基金资助项目(2013FB134)
