水通道蛋白4小分子抑制剂的筛选和验证

doi:10.13481/j.1671-587X.20210234

Abstract

Abstract: Objective

To screen the small molecules that can inhibit the binding of neuromyelitis optica（NMO）-IgG to aquaporin 4（AQP4） by high-throughput screening method.

Methods

The FRT-AQP4 cells were divided into negative control group （no small molecule compounds） and screening group， and the horseradish peroxidase（HRP） activity was detected by chemiluminescence. The V79-AQP4 cells are divided into inhibitor group （neosutchuenenine） and DMSO group.The fluorescence intensity of each group was detected by immunofluorescence method， the water permeabilitise of cells in varions groups were detected by fluorescence quenching method， and complement dependent cytotoxicity was detected by CellTiter-Glo^? luminescence method；the cell activities were detected by water solubility tetrazole-1（WST-1） method，and the apoptosis was detected by enzyme linked immunosorbent assay（ELISA） method.

Results

Compared with negative control group， the HRP activity in screening group was significantly increased（P<0.01）. Compared with DMSO group， the red fluorescence intensity in inhibitor group was significantly reduced， while the green fluorescence did not change significantly； compared with DMSO group， the water permeability in inhibitor group did not change significantly（P>0.05）； compared with DMSO group， the complement-dependent cytotoxicity in inhibitor group was significantly reduced （P<0.01）； compared with DMSO group， there was no significant changes in cell activities and apoptosis in inhibitor group（P>0.05）.

Conclusion

A high-throughput screening method for AQP4 small molecule inhibitor is successfully established， and neosutchuenenine， a small molecule inhibitor with blocking effect， is screened out.

Key words: aquaporin, neuromyelitis optica, small molecule inhibitors, high throughput screening

CLC Number:

Q503

Nan WANG,Jian GUO,Bing YAN,Qing LIU,Lei ZHANG,Huijing XU,Miao LI,Meiyan SUN. Screening and validation of small molecule inhibitors of aquaporin 4[J].Journal of Jilin University(Medicine Edition), 2021, 47(2): 511-518.

Figures/Tables 7

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Screening and validation of small molecule inhibitors of aquaporin 4

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