Journal: Annals of Translational Medicine

Article Title: Idebenone reduces sepsis-induced oxidative stress and apoptosis in hepatocytes via RAGE/p38 signaling

doi: 10.21037/atm-22-5758

Figure Lengend Snippet: IDE suppressed the expressions of RAGE/p38 signaling. (A,B) The expressions of RAGE and p-p38/p38 were detected using western blot. ***P<0.001 vs. control group; ### P<0.001 vs. sepsis group. RAGE, receptor for advanced glycation end products; p, phosphorylated; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IDE, idebenone.

Article Snippet: Subsequently, the PVDF membranes were blocked using 5% non-fat milk or 5% bovine serum albumin (BSA) and subsequently cultivated with primary antibodies targeting Bcl-2 (ab196495; 1:1,000; Abcam, Shanghai, China), Bax (ab32503; 1:1,000; Abcam), cleaved caspase3 (19677-1-AP; 1:10,000; Proteintech, Wuhan, China), RAGE (ab216329; 1:1,000; Abcam), phosphorylated (p)-p38 (ab195049; 1:1,000; Abcam), p38 (ab170099; 1:1,000; Abcam), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab181602; 1:10,000; Abcam) overnight at 4 °C.

Techniques: Western Blot, Control

Journal: Annals of Translational Medicine

Article Title: Idebenone reduces sepsis-induced oxidative stress and apoptosis in hepatocytes via RAGE/p38 signaling

doi: 10.21037/atm-22-5758

Figure Lengend Snippet: IDE alleviated LPS-induced hepatocyte activity damage and suppressed the expressions of RAGE/p38 signaling. (A) The viability of hepatocytes was detected using CCK-8. (B) The viability of LPS-induced hepatocytes was detected using CCK-8. (C,D) The expressions of RAGE and p-p38/p38 were detected using western blot. ***P<0.001 vs. control group; # P<0.05, ### P<0.001 vs. LPS group. LPS, lipopolysaccharide; ns, not significant; RAGE, receptor for advanced glycation end products; p, phosphorylated; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IDE, idebenone; CCK-8, cell counting kit-8.

Article Snippet: Subsequently, the PVDF membranes were blocked using 5% non-fat milk or 5% bovine serum albumin (BSA) and subsequently cultivated with primary antibodies targeting Bcl-2 (ab196495; 1:1,000; Abcam, Shanghai, China), Bax (ab32503; 1:1,000; Abcam), cleaved caspase3 (19677-1-AP; 1:10,000; Proteintech, Wuhan, China), RAGE (ab216329; 1:1,000; Abcam), phosphorylated (p)-p38 (ab195049; 1:1,000; Abcam), p38 (ab170099; 1:1,000; Abcam), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab181602; 1:10,000; Abcam) overnight at 4 °C.

Techniques: Activity Assay, CCK-8 Assay, Western Blot, Control, Cell Counting

Journal: Annals of Translational Medicine

Article Title: Idebenone reduces sepsis-induced oxidative stress and apoptosis in hepatocytes via RAGE/p38 signaling

doi: 10.21037/atm-22-5758

Figure Lengend Snippet: IDE alleviated LPS-induced hepatocyte activity damage and apoptosis via RAGE/p38 signaling. (A) The mRNA expression of RAGE was detected using RT-qPCR. ***P<0.001 vs. Ov-NC group. (B) The protein expression of RAGE was detected using western blot. ***P<0.001 vs. Ov-NC group. (C) The viability of LPS-induced hepatocytes was detected using CCK-8. (D,E) The apoptosis of LPS-induced hepatocytes was detected using TUNEL. (F,G) The expressions of apoptosis-related proteins were detected using western blot. ***P<0.001 vs. control group; ### P<0.001 vs. LPS group; $ P<0.05, $$ P<0.01, $$$ P<0.001 vs. LPS + IDE + Ov-NC group. RAGE, receptor for advanced glycation end products; mRNA, messenger RNA; Ov, overexpression; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; TUNEL, terminal deoxynucleotidyl transferase-mediated nick-end labeling; DAPI, 4',6-diamidino-2-phenylindole; IDE, idebenone; RT-qPCR, real-time quantitative polymerase chain reaction; CCK-8, cell counting kit-8.

Article Snippet: Subsequently, the PVDF membranes were blocked using 5% non-fat milk or 5% bovine serum albumin (BSA) and subsequently cultivated with primary antibodies targeting Bcl-2 (ab196495; 1:1,000; Abcam, Shanghai, China), Bax (ab32503; 1:1,000; Abcam), cleaved caspase3 (19677-1-AP; 1:10,000; Proteintech, Wuhan, China), RAGE (ab216329; 1:1,000; Abcam), phosphorylated (p)-p38 (ab195049; 1:1,000; Abcam), p38 (ab170099; 1:1,000; Abcam), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab181602; 1:10,000; Abcam) overnight at 4 °C.

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, TUNEL Assay, Control, Over Expression, Negative Control, End Labeling, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Annals of Translational Medicine

Article Title: Idebenone reduces sepsis-induced oxidative stress and apoptosis in hepatocytes via RAGE/p38 signaling

doi: 10.21037/atm-22-5758

Figure Lengend Snippet: IDE alleviated LPS-induced oxidative stress and inflammatory release in hepatocytes via RAGE/p38 signaling. (A) The expression of MDA was detected using an MDA assay kit. (B) The expression of SOD was detected using a SOD assay kit. (C) The expression of GSH-Px was detected using a GSH-Px assay kit. (D) The ROS content was detected using DCFH-DA. The hepatocytes were stained with DCFH-DA. Magnification, ×200. (E) The levels of TNF-α, IL-1β, and IL-6 were detected using ELISA. ***P<0.001 vs. control group; ### P<0.001 vs. LPS group; $$$ P<0.001 vs. LPS + IDE + Ov-NC group. MDA, malondialdehyde; LPS, lipopolysaccharide; Ov, overexpression; NC, negative control; RAGE, receptor for advanced glycation end products; SOD, superoxide dismutase; GSH-Px, glutathione peroxidase; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; IL-6, interleukin-6; IDE, idebenone; ROS, reactive oxygen species; DCFH-DA, dichloro-dihydro-fluorescein diacetate; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Subsequently, the PVDF membranes were blocked using 5% non-fat milk or 5% bovine serum albumin (BSA) and subsequently cultivated with primary antibodies targeting Bcl-2 (ab196495; 1:1,000; Abcam, Shanghai, China), Bax (ab32503; 1:1,000; Abcam), cleaved caspase3 (19677-1-AP; 1:10,000; Proteintech, Wuhan, China), RAGE (ab216329; 1:1,000; Abcam), phosphorylated (p)-p38 (ab195049; 1:1,000; Abcam), p38 (ab170099; 1:1,000; Abcam), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab181602; 1:10,000; Abcam) overnight at 4 °C.

Techniques: Expressing, Multiple Displacement Amplification, Staining, Enzyme-linked Immunosorbent Assay, Control, Over Expression, Negative Control