rhFGF4在大肠杆菌中的表达、纯化及其对高糖损伤的人脐静脉内皮细胞的保护作用

doi:10.3969/j.issn.2095-9400.2021.07.001

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Abstract Objective: To prepare and identify recombinant human fibroblast growth factor 4 (rhFGF4) protein and observe the protective effect of rhFGF4 on human umbilical vein endothelial cells (HUVEC) model with high glucose damage. Methods: The recombinant plasmid pET3d-hFGF4 was constructed, and the rhFGF4 protein was efficiently expressed by E.coli expression system. Most soluble objective proteins are expressed in the form of inclusion, including the cleaning, degeneration and restoration and purification of heparin column.The MTT method was used to detect the biological activity of rhFGF4 protein in promoting cell proliferation,the cell damage model was established and the phosphorylation expression of NF-κB, JNK and p38 was detected by Western blot technology. Results: The recombinant plasmid pET3D-HFGF4 was successfully constructed.SDS-PAGE electrophoresis and Western blot identification results showed that rhFGF4 protein was successfully expressed in E.coli. It was also confirmed that rhFGF4 protein repaired HUVEC cells damaged by high glucose and inhibited their apoptosis by preventing the phosphorylation of NF-κB, JNK and p38, and reduced their MDA and ROS content. Conclusion: This study has successfully obtained high purity and biologically active rhFGF4 protein, which provides a useful tool for further research on the role of this protein in vascular diseases caused by high glucose and related mechanisms.

Key words： rhFGF4 on-column refolding human umbilical vein endothelial cells oxidative stress apoptosis

Received: 30 December 2020

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	ZHANG Yu
	WU Fenfang
	ZHANG Meng
	TIAN Haishan
	LI Xiaokun

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https://xb.wmu.edu.cn/EN/10.3969/j.issn.2095-9400.2021.07.001 OR https://xb.wmu.edu.cn/EN/Y2021/V51/I7/517

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