Inhibitory role of Ulinastatin in TNF-α induced hyperpermeability of endothelial cells via Rho/ROCK pathway

doi:10.3877/cma.j.issn.2096-1537.2018.02.013

Abstract

Abstract:

Objective

To investigate the role of Ulinastatin (UTI) on the hyper-permeability of human umbilical vein endothelial cell by TNF-α and possible underlying mechanism.

Methods

Human umbilical vein endothelial cell (EA.hy926) were cultured in vitro and exposed to UTI and tumor necrosis factor alpha (TNF-α) respectively. The permeability of EA.hy926 cells were detected by a transwell chamber system. Immunofluorescence was used to assay the expression of p-MYPT1. The expression of the key molecules related to endothelial permeability (RhoA, ROCK2, MYPT1, p-MYPT1 and VE-cadherin) were detected by immunocytochemistry assays and western-blot.

Results

Compared with the control group, the permeability of cell exposed to TNF-αwas increased significantly, the expression of p-MYPT1 was higher, but UTI could attenuate the phenomena. The immunocytochemistry assays showed that the expression of RhoA and ROCK2 was significantly upregulated in cells treated with TNF-α; however, UTI could inhibit the high expression of these two proteins. Western blots showed that the expression of p-MYPT1, RhoA and ROCK2 were significantly higher (P<0.05) and VE-cadherin was significantly lower (P<0.05) after incubation with TNF-α, but UTI treatment could moderate the above changes.

Conclusion

The serum from septic patients induced hyperpermeability of EA.hy926 cells can be attenuated by ulinastatin in a dose-dependent manner.

Key words: Human umbilical vein endothelial cell line EA.hy926, Ulinastatin, High permeability, Tumor necrosis factor alpha, Rho/ROCK pathway

Siyi Liu, Fu Wei, Shuke Liu, Li Luo, Shan Xu, Dan Zhang. Inhibitory role of Ulinastatin in TNF-α induced hyperpermeability of endothelial cells via Rho/ROCK pathway[J]. Chinese Journal of Critical Care & Intensive Care Medicine(Electronic Edition), 2018, 04(02): 170-175.

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URL: https://icu.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2096-1537.2018.02.013

https://icu.cma-cmc.com.cn/EN/Y2018/V04/I02/170

Figures/Tables 4

References 23

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组别	相对通透性（吸光值A值）
1 U/ml UTI＋TNF-α组	1.854±0.064
10 U/ml UTI＋TNF-α组	1.500±0.081^b
100 U/ml UTI＋TNF-α组	1.234±0.013^b
1000 U/ml UTI＋TNF-α组	1.654±0.155
TNF-α组	2.097±0.308^a
正常对照组	1.346±0.038

组别	相对通透性（吸光值A值）
1 U/ml UTI＋TNF-α组	1.854±0.064
10 U/ml UTI＋TNF-α组	1.500±0.081^b
100 U/ml UTI＋TNF-α组	1.234±0.013^b
1000 U/ml UTI＋TNF-α组	1.654±0.155
TNF-α组	2.097±0.308^a
正常对照组	1.346±0.038

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