Fig 2: Effect of glycyrrhizin on the mRNA expression of kidney injury markers and serum lactate dehydrogenase (LDH). (A-C) Relative expressions of KIM-1, NGAL and IL-18 mRNA were measured using RT-PCR. The mRNA expression of kidney injury markers was significantly higher in the PC-AKI group compared to the controls, whereas it was significantly lower in the PC-AKI with glycyrrhizin group. (D) Compared to the control group, serum LDH levels significantly increased in PC-AKI, whereas they decreased with glycyrrhizin pretreatment. Results were expressed as means ± SEMs. N = 8 for each group. Statistical significance: *P < 0.05 and ***P < 0.001 Con vs PC-AKI and Con vs PC-AKI + GL, #P < 0.05 and ##P < 0.01 PC-AKI vs GL + PC-AKI. Abbreviation: PC-AKI, post-contrast acute kidney injury; GL, glycyrrhizin; LDH, lactate dehydrogenase.

Fig 3: Knocking down HMGB1 inhibits the mRNA expression of kidney injury markers after contrast media exposure. (A-C) The mRNA expressions of KIM-1, NGAL and IL-18 mRNA were measured using RT-PCR. Similar to pro-inflammatory cytokines, the mRNA expression of kidney injury markers did not significantly increase after cells were exposed to contrast media or contrast media + glycyrrhizin after knocking down HMGB1. Results were expressed as means ± SEMs for the three independent experiments, which were each performed in duplicate. Statistical significance: **P < 0.01 Con vs siHMGB1 + CM + GL. #P < 0.05, ##P < 0.01 and ###P < 0.001, CM vs CM + GL, CM vs siHMGB1 + CM, CM vs siHMGB1 + CM + GL. Abbreviation: NC, negative control; CM, contrast media; GL, glycyrrhizin.

Fig 4: Renal injury markers after repeated CT contrast agent administration. A-D: Real-time polymerase chain reaction was used to measure the mRNA expression of KIM-1 and NGAL (A, B). An ELISA was used to quantify protein levels of KIM-1 and NGAL (C, D). Regardless of time intervals, the repeated exposure to the CT contrast agent did not affect both mRNA and protein levels of KIM-1 and NGAL. Results are presented as the mean ± standard deviation. KIM-1 = kidney injury molecule-1, NGAL = neutrophil gelatinase-associated lipocalin, ELISA = enzyme-linked immunosorbent assay, NC = negative control, PC = positive control, CCT_2, 4, and 24 hr = an additional dose of CT contrast agent within 2, 4, or 24 hours after the single-dose administration of CT contrast agent, n.s. = no significance

Fig 5: Late renal denervation (RDN) intervention does not influence renal NLR family pyrin domain containing 3 (NLRP3) inflammasome synthesis. A, Representative images of renal interstitial fibrosis in sham RDN (sham‐RDN) (left) and RDN‐treated (right) animals stained with Masson trichrome and quantification of renal interstitial fibrosis. B, Representative images of renal medium‐large arteries and quantification of renal perivascular fibrosis. C, Representative images and quantification of glomerular size. D, Representative images and quantification of cast abundance in renal tubules. E, Circulating kidney injury marker‐1 (KIM‐1). F, Circulating blood urea nitrogen (BUN). G, Circulating albumin. H, Polymerase chain reaction (PCR) quantification of profibrotic gene expression in renal tissue. I, PCR quantification of proinflammatory and NLRP3 inflammasome‐related gene expression in renal tissue. Circles inside bars indicate sample size. Data in (A) through (D) were analyzed with 2‐tailed Mann–Whitney U test. Data in panels (E) through (I) were analyzed with unpaired 2‐tailed Student t test. Data are presented as mean±SEM. ASC indicates apoptosis‐associated speck‐like protein containing a caspase recruitment domain; Casp‐1, caspase 1; Col1a1, collagen type 1 alpha 1 chain; Col3a1, collagen type 3 alpha 1 chain; FC, fold change; IL, interleukin; NGAL, neutrophil gelatinase–associated lipocalin; TGFβ1, transforming growth factor beta 1; and TNFα, tumor necrosis factor alpha.