“复活”的人尿酸氧化酶在人肝细胞中的表达、定位和代谢功能。

Expression, localization and metabolic function of "resurrected" human urate oxidase in human hepatocytes.

机构信息

School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China.

出版信息

Int J Biol Macromol. 2021 Apr 1;175:30-39. doi: 10.1016/j.ijbiomac.2021.01.163. Epub 2021 Jan 26.

DOI:10.1016/j.ijbiomac.2021.01.163

Abstract

A high serum uric acid (SUA) concentration is associated with hyperuricemia (HUA) and gout. In order to obtain long-acting therapeutic effect, correction of purine metabolism at genetic level is advantageous. For this purpose, we expressed three "human-like" urate oxidases in human hepatocytes (HL-7702) by lentivirus-mediated transduction. Enzymatic assay revealed that the recombinant urate oxidases expressed in HL-7702 cells were functionally active. Electron microscopy study showed that the recombinant enzymes were localized to peroxisome and formed distinct crystalloid core structures as in other mammal cells. Although similar rate of uric acid degradation was observed for all recombinant urate oxidases, HL-7702-pLVX-UOX cells and HL-7702-pLVX-UOX cells retained more cell viability compared with HL-7702-pLVX-UOX at high uric acid level. This study provides a new direction for the treatment of gout and hyperuricemia.

摘要

血清尿酸（SUA）浓度升高与高尿酸血症（HUA）和痛风有关。为了获得长效治疗效果，纠正嘌呤代谢的基因水平是有利的。为此，我们通过慢病毒介导的转导在人肝细胞（HL-7702）中表达了三种“类人”尿酸氧化酶。酶活性分析显示，在 HL-7702 细胞中表达的重组尿酸氧化酶具有功能活性。电子显微镜研究表明，重组酶定位于过氧化物酶体，并形成与其他哺乳动物细胞相似的明显结晶核心结构。尽管所有重组尿酸氧化酶都观察到尿酸降解的相似速率，但与 HL-7702-pLVX-UOX 细胞相比，在高尿酸水平下，HL-7702-pLVX-UOX 细胞和 HL-7702-pLVX-UOX 细胞保留了更高的细胞活力。这项研究为痛风和高尿酸血症的治疗提供了新的方向。