2019•Chin J Cell Stem Cell (Electronic Edition)Requires access
Effect of TLR4 on LPS-induced bronchial epithelial 16HBE cell injury
Guangfu Shen, Changqin Luo, Bo Huang, Jing Bi, Xuan Zhang, Rui Yu, Jun Yang, Jun Xie
Open publisher page 0 citations
Abstract
Objective
To investigate the effect of TLR4 on LPS-induced bronchial epithelial 16HBE cell injury and its mechanism.
Methods
Three siRNA-TLR4-1, siRNA-TLR4-2 and siRNA-TLR4-3 were transfected into 16HBE cells, and the best interference sequence was screened for experiment. The experiment was divided into a control group (untreated) , LPS group (treated with 50 μg/ml LPS) , LPS + siNC group (treated with 50 μg/mL LPS after transfection of siRNA- NC) and LPS + siTLR4 group (treated with 50 μg/ml LPS after transfection of siRNA-TLR4) . The expression of TLR4, IL-6 and TNF-α was detected by RT-PCR, cell viability was detected by MTT, apoptotic rate was detected by flow cytometry, and the expression of Bcl-2, Bax, Cleaved Caspase- 3, NF-κBp65 and IκBα proteins were tested by Western Blot. One-way analysis of variance was used for comparison between groups. SNK-q was used for multiple comparisons between groups, and independent sample t test was used for comparison between the two groups.
Results
The TLR4 mRNA (2.05±0.12 vs 3.28±0.15) and protein expression (0.38±0.03 vs 0.77±0.05) in the LPS group and the LPS+siTLR4-2 group were statistically significant (t = 11.091, 11.585, P 0.05) .
Conclusion
Downregulation of TLR4 can alleviate 16HBE cell injury by inhibiting activation of NF-κB pathway, inhibiting LPS-induced apoptosis and inflammation.
Key words:
Toll-like Receptor 4; Lipopolysaccharide; Bronchial epithelial cells; Inflammation; Apoptosis