四环素对纳米细菌感染的人肾小管上皮细胞HK-2中CaSR、Claudin-14表达的影响
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篇名: | 四环素对纳米细菌感染的人肾小管上皮细胞HK-2中CaSR、Claudin-14表达的影响 |
TITLE: | |
摘要: | 目的:研究四环素对纳米细菌(NB)感染的人肾小管上皮细胞HK-2中钙敏感受体(CaSR)、紧密连接蛋白14(Claudin-14)表达的影响,为抑制肾结石的形成提供参考。方法:将HK-2细胞分为空白对照组、NB组和四环素组(NB+四环素),每组3个复孔,各组分别加入相应NB和药物后共培养24 h。采用透射电子显微镜观察各组细胞的形态和超微结构变化,运用实时荧光定量聚合酶链式反应(qRT-PCR)和Western blot法分别检测细胞中CaSR、Claudin-14 mRNA和蛋白的表达水平。结果:与空白对照组比较,NB组细胞膜结构破损,胞核裸露在外,微绒毛基本消失,线粒体数量较少,结构肿胀,细胞中CaSR、Claudin-14 mRNA和蛋白表达水平明显升高(P<0.05);四环素组细胞膜结构完整,微绒毛结构稍紊乱,局部绒毛消失,部分线粒体结构肿胀不明显,细胞中CaSR、Claudin-14 mRNA和蛋白表达水平明显升高(P<0.05)。与NB组比较,四环素组细胞中CaSR、Claudin-14 mRNA和蛋白表达水平明显降低(P<0.05)。结论:NB可能通过上调HK-2细胞中CaSR、Claudin-14 mRNA和蛋白的表达促进肾结石的形成,而四环素可能通过作用于NB降低CaSR、Claudin-14 mRNA和蛋白的表达抑制肾结石的形成。 |
ABSTRACT: | OBJECTIVE: To study the effects of tetracycline on the expression of calcium-sensing receptor (CaSR) and Claudin-14 in nanobacteria-infected (NB) human renal tubular epithelial cells HK-2, and to provide reference for inhibiting the formation of kidney stone. METHODS: HK-2 cells were divided into blank control group, NB group and tetracycline group (NB+tetracycline), with 3 complex holes in each group. They were given relevant nanobacteria and drug for 24 h. Morphology and ultrastructural changes of cells were observed by TEM. mRNA and protein expression of CaSR and Claudin-14 were measured by real-time quantitative PCR (qRT-PCR) and Western blot assay. RESULTS: Compared with blank control group, the damaged structure of cell membrane, exposed nucleus, disappeared microvilli, small number of mitochondria and swollen structure were found in NB group; mRNA and protein expression of CaSR and Claudin-14 were increased significantly (P<0.05). The complete structure of cell membrane, disordered structure of microvilli, disappeared local villi and not obvious swollen of some mitochondrial structure were found in tetracycline group; mRNA and protein expression of CaSR and Claudin-14 were increased significantly (P<0.05). Compared with NB group, mRNA and protein expression of CaSR and Claudin-14 were decreased significantly in tetracycline group (P<0.05). CONCLUSIONS: NB may improve the formation of kidney stones by up-regulating the mRNA and protein expression of CaSR and Claudin-14 in HK-2 cells, while tetracycline may reduce the mRNA and protein expression of CaSR and Claudin-14 by anti-NB effect so as to inhibit kidney stone formation. |
期刊: | 2018年第29卷第19期 |
作者: | 申茂磊,高存祥,郑丽英,王勤章,徐浩,郝志强,傅家豪,钱彪 |
AUTHORS: | SHEN Maolei,GAO Cunxiang,ZHENG Liying,WANG Qinzhang,XU Hao,HAO Zhiqiang,FU Jiahao,QIAN Biao |
关键字: | 四环素;纳米细菌;人肾小管上皮细胞HK-2;钙敏感受体;紧密连接蛋白14 |
KEYWORDS: | Tetracycline; Nanobacteria; Human renal tubular epithelial cells HK-2; CaSR; Claudin-14 |
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