Table 1.
Primer sequences.
Fig 1.
Combined treatment with Bud and NAC provides stronger protection against ALI.
A. HE staining for lung pathological changes. B. Evans blue staining for alveolar permeability. C. Detection of ratio of wet to dry weight of the lung assessment for pulmonary edema. D. TUNEL kit for apoptosis. E-F. The expression of focal death-related proteins was detected by Western blot. G. ELISA for inflammatory factor levels. caspase -1: active-caspase -1; IL-1β: mature- IL-1β; IL-18: mature-IL-18. Vs NC, ** p < 0.01; Vs ALI, △p < 0.05, △△p < 0.01; Vs Bud+NAC #p < 0.05, ##p < 0.01.
Fig 2.
Combined treatment with Bud and NAC attenuates LPS-induced alterations in proliferative viability, apoptosis, scorching and inflammatory response in human bronchial epithelial cells.
A. CCK-8 analysis of cell proliferation activity. B. Flow cytometry for apoptosis. C. Western blot for scorch death-related protein expression. D. ELISA for inflammatory factor levels. caspase -1: active-caspase -1; IL-1β: mature- IL-1β; IL-18: mature-IL-18. Vs NC, ## p < 0.01; Vs LPS, ** p < 0.01; Vs LPS+Bud+NAC, △△p < 0.01.
Fig 3.
Combined treatment with Bud and NAC exerts ALI protective effects through upregulation of miR-381 expression.
A-C. RT–qPCR to detect miR-381 expression in BEAS-2B cells. D. CCK-8 analysis of cell proliferation activity. E. Flow cytometry for apoptosis. F. Western blot for scorch death-related protein expression. G. ELISA for inflammatory factor levels. caspase -1: active-caspase -1; IL-1β: mature- IL-1β; IL-18: mature-IL-18. Vs NC, ## p < 0.01; Vs LPS, ** p < 0.01;; Vs LPS+Bud+NAC, △△p < 0.01.
Fig 4.
miR-381 targets and regulates NLRP3 expression.
A. Targeted binding sites for miR-381 and NLRP3 are predicted by http://www.microrna.org/. B. Validation of the targeting relationship between miR-381 and NLRP3 with dual luciferase reporter genes. C. Western blot for NLRP3 expression. D. Immunohistochemical detection of NLRP3 expression in rat lung tissues. Vs NC, ** p < 0.01.
Fig 5.
Bud and NAC attenuate LPS-induced alterations in the proliferative viability, apoptosis, focal death and inflammatory response of human bronchial epithelial cells via miR-381/NLRP3.
A and D. western blot for the expression of the NLRP3 and pyrodeath-related protein; B. CCK-8 analysis of cell proliferation viability. C. Flow cytometry for apoptosis. E. ELISA for inflammatory factor levels. caspase -1: active-caspase -1; IL-1β: mature- IL-1β; IL-18: mature-IL-18. Vs NC, ** p < 0.01; Vs LPS, △△p < 0.01; Vs LPS+Bud+NAC #p < 0.05, ##p < 0.01.