Penetration ability and biological activity of fusion protein PTD4-Cu, Zn-SOD in human astrocytes in vitro by immunofluorescence test

Lihua Meng; Rongliang Xue; Xiaoming Lei; Luming Zhen; Lanying Wei

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Penetration ability and biological activity of fusion protein PTD4-Cu, Zn-SOD in human astrocytes in vitro by immunofluorescence test

VernacularTitle:免疫荧光法检测融合蛋白PTD4-Cu,Zn-SOD在人星形胶质细胞中穿膜能力及保护作用
Author: Lihua MENG ¹ ; Rongliang XUE ¹ ; Xiaoming LEI ¹ ; Luming ZHEN ¹ ; Lanying WEI ¹
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1. Department of Anesthesiology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China
Publication Type:Journal Article
Keywords: PTD-SOD fusion protein; penetration ability; Immunofluorescence
From: Journal of Xi'an Jiaotong University(Medical Sciences) 2023;44(2):309-314
CountryChina
Language:Chinese
Abstract: 【Objective】 To observe the penetration and biological activity of PTD4-Cu, Zn-SOD into human astrocytes and whether it can mitigate hypoxia damages. 【Methods】 ①Immunohistochemistry and fluorescence test: We labeled the Cu, Zn-SOD by a monoclonal antibody, combined it with the fluorescent secondary antibody labeled with fluorescein isothiocyanate (FITC) to observe the effect of transduced PTD4-Cu, Zn-SOD on the viability of human astrocytes. ② The experimental group: After hypoxic damage model, the cells were divided into three groups: blank control, group Cu, Zn-SOD, and group PTD4-Cu, Zn-SOD. Group blank was added with DMEM medium (excluding serum) as control; DMEM medium was added to the other two for one hour (excluding serum) with its fusion proteins (Cu, Zn-SOD and PTD4 -Cu, Zn-SOD) with the final concentration of 2 μmoL/L. After the intervention, we used SOD and MDA test kits to observe PTD4-Cu, Zn-SOD and Cu, Zn-SOD in astrocytes after fusion protein intervention. 【Results】 The PTD4-Cu, Zn-SOD fusion protein could have aggregation distribution in the nucleus by FITC fluorescently labeled. After the intervention, it could increase the SOD activity in astrocytes in group PTD4-Cu, Zn-SOD and group Cu, Zn-SOD compared with control group, but the SOD activity was more obvious in the fusion proteins PTD4-Cu, Zn-SOD group. And the dose of MDA was reduced in group PTD4-Cu, Zn-SOD compared with group Cu, Zn-SOD and control group. 【Conclusion】 PTD4-Cu, Zn-SOD fusion protein can transcellular membrane of human astrocytes. The fusion protein PTD4-Cu, Zn-SOD can increase the SOD activity and reduce the content of MDA by human astrocytes from hypoxia injury.