Fig 1: Nrf2 binds to the PHB2 promoter and enhances gene transcription. (A) Input DEGs of GSE95233 into ChEA3 (https://maayanlab.cloud/chea3/) to obtain the downstream of Nrf2. (B) Dual‐luciferase reporter system was used to test Nrf2 binds to the PHB2 promoter region. The thymidine kinase promoter‐renilla luciferase (pRL‐TK) vector was transfected as an internal reference to correct the transfection efficiency. (C–E) The relative levels of complex IV, ND‐1, and COX‐1 were used to reflect the mtDNA copy number. (F) ATP content was measured in A549 cells treated with Nrf2‐OE plasmid and PHB2 siRNA. Experiments were repeated at least three times *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns denotes not significant.
Fig 2: Decreased Nrf2 expression in S. aureus‐induced sepsis (SA‐sepsis) patients. (A) Gene expression of Nrf2 in 22 healthy samples and 102 sepsis samples were obtained from GSE95233. (B) Nrf2 protein levels in serum from control (n = 22) and SA‐sepsis (n = 11) patients. (C and D) Correlation between serum Nrf2 content and APACHE II/SOFA score in SA‐sepsis. (E and F) Levels of PCT and PaO2/FiO2 from the clinical data of patients. (G) The survival probability of patients within 28 days after admission to the ICU was analyzed. **p < 0.01.
Fig 3: Effects of coixol on protein expression of phosphorylated NF-κB p65, phosphorylated p38, iNOS, RAGE, and Nrf2. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. iNOS, inducible nitric oxide synthase; NF-κB, nuclear factor kappa B; NGF, nerve growth factor; RAGE, receptor of advanced glycation end product; SD, standard deviation.
Fig 4: Nrf2 deficiency aggravates SA‐ALI with mitochondrial dysfunction. (A) Pathological changes in lung tissue due to sepsis and Nrf2 deficiency. (magnification, ×100 and ×400, bar = 200 and 100 μm; n = 5). (B) Lung injury scores based on the pathological results. (C and D) Cell counts and protein concentration in BALF from mice. (E and F) Levels of serum SOD and LDH from Nrf2−/− mice. (G–I) The relative mRNA levels of complex IV, COX‐1, and ND‐1 were used to reflect the mtDNA copy number. (J) PHB2 mRNA level in lung tissue. Experiments were repeated at least three times. *p < 0.05, **p < 0.01, ****p < 0.0001, ns denotes not significant.
Fig 5: Nrf2 inhibition exacerbates mitochondrial dysfunction in vitro. (A) In vitro, total proteins were collected from LTA‐treated A549 cells at concentrations of 10 μg/mL for 30 min, 2 h, and 4 h, respectively. (B) A549 cells were treated separately by LTA at different concentration gradients for 4 h. β‐Actin were used as internal reference proteins. (C) Nrf2 level in cells after ML385 and LTA treatment was detected using western blot. (D and E) Cell vitality and ATP concentration was measured in A549 cells. (F) MMP was recorded using JC‐1 staining (magnification, ×400, bar = 100 μm; n = 3). Experiments were repeated at least three times. *p < 0.05, **p < 0.01, ns denotes not significant.
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