Bai Jijia,Shi Guocui,Ma Shenmao,et al.TNF-α downregulates the expression of tight junction proteins ZO-1 and Claudin-5 in rat pulmonary microvascular endothelial cells[J].Journal of Chongqing Medical University,2024,49(2):125-131.
Bai Jijia,Shi Guocui,Ma Shenmao,et al.TNF-α downregulates the expression of tight junction proteins ZO-1 and Claudin-5 in rat pulmonary microvascular endothelial cells[J].Journal of Chongqing Medical University,2024,49(2):125-131. DOI: 10.13406/j.cnki.cyxb.003423.
TNF-α downregulates the expression of tight junction proteins ZO-1 and Claudin-5 in rat pulmonary microvascular endothelial cells
To investigate the effect of tumor necrosis factor-α(TNF-α) on the expression of tight junction proteins ZO-1 and Claudin-5 in rat pulmonary microvascular endothelial cells and to explore the injury mechanism of TNF-α on pulmonary vascular barrier in rats.
Methods
2
Rat pulmonary microvascular endothelial cells cultured
in vitro
were divided into control group,and TNF-α 24 h,48 h,and 72 h groups(TNF-α 10 mg/L). The ultrastructure of rat pulmonary microvascular endothelial cells was observed by a transmission electron microscope. The proliferation inhibition rate and apoptosis rate of cells in each group were determined by MTT assay and flow cytometry. The distribution of ZO-1 and Claudin-5 was determined by the immunofluorescence method. The mRNA and protein expression levels of ZO-1 and Claudin-5 in cells of each group were determined by real-time fluorescence quantitative polymerase chain reaction and Western blot.
Results
2
Endothelial cells in the control group contained a large number of mitochondria and endoplasmic reticulum,with a normal structure. Cells in the TNF-α groups showed a decrease in the number of mitochondria,mitochondrial swelling,cristae lysis,karyopyknosis,and an increase in apoptotic cells,with a significant time-dependent increase in the proliferation inhibition rate and apoptosis rate compared with the control group(all
P
<
0.05). ZO-1 and Claudin-5 were distributed at the junction of endothelial cells,showing linear fluorescence. Compared with the control group,the TNF-α groups showed a significant decrease in the transcription and expression levels of ZO-1 and the transcription level of Claudin-5(
P
<
0.05); the TNF-α 48 h and 72 h groups showed a
significant decrease in the protein expression level of Claudin-5(
P
<
0.05); the TNF-α 24 h group showed no significant difference(
P
>
0.05).
Conclusion
2
TNF-α downregulates the expression of ZO-1 and Claudin-5 in rat pulmonary microvascular endothelial cells,induces cell injury and apoptosis,destroys the lung barrier,and leads to lung injury.
关键词
Keywords
references
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