Fig 2: Effects of Cordyceps cicadae (C. cicadae) and resveratrol (RES) on tubular epithelial cell (TEC) injury induced by angiotensin II (AngII). (a) The level of urinary kim-1 was measured with enzyme-linked immunosorbent assay (ELISA) in the rats in all the groups. C. cicadae: spontaneously hypertensive rats (SHRs) treated with 4 g/kg/d C. cicadae; RES: SHRs treated with 40 mg/kg/d resveratrol. (b, c) Levels of kim-1 and NGAL were measured with ELISA in culture supernatants of TECs treated with 10% CSM and/or 10 μM EX527 in the presence or absence of 10−7 mM AngII. (d–g) Levels of kim-1 and NGAL were measured with ELISA in culture supernatants of TECs. The CSM and RES treatments reduced the secretion of kim-1 and NGAL in the TECs after exposure to AngII, and EX527 reversed the beneficial effect of CSM. EP: TECs treated with 12.5 μg/ml ergosterol peroxide. RES: TECs treated with 25 μM resveratrol. #p < 0.05 and ##p < 0.01, as compared with the WKY or control TECs. ∗p < 0.05 and ∗∗p < 0.01, as compared with SHRs or TECs exposed to AngII. ∧p < 0.05, as compared with TECs exposed to AngII plus CSM.

Fig 3: CMS attenuated AngII-associated injury and inhibited ECM accumulation in rat renal tubular epithelial cells. (A) NRK-52E cells were exposed to various concentrations of AngII for 24 and 72 h. (B) Effects of CMS on viability for 72 h. (C) Effects of RES on NRK-52E cell viability for 72 h. (D,E) Effect of CMS on NRK-52E cell injury. The level of cell injury was examined by KIM-1 and NGAL ELISA kits. (F) Effect of CMS on the expression of Col-1 and α-SMA. (G,H) The relative intensities of fibrosis-related protein in cells were calculated after normalization against GAPDH. Data are expressed as the mean ± SD of three separate experiments. # p < 0.05, ## p < 0.01 compared with control cells. *p < 0.05, **p < 0.01, compared with the model group.

Fig 4: TMP alleviates renal tubular pathological injury and improves renal function in rats with renal I/R. (A) Photomicrographs of periodic acid-Schiff staining showing typical renal tubular injury in Sham, I/R and I/R+TMP rats, red boxes highlight the corresponding areas of the high-power images. (B) Summarized data showing the renal tubular injury score in each group. (C) The ELISA results showing the level of KIM-1 in each group. (D and E) SCr and BUN levels in each group were measured by a biochemical analyzer. (F and G) The ELISA results showing the levels of the inflammatory cytokines TNF-a and IL-6 in each group. *P<0.05 vs. Sham rats; #P<0.05 vs. I/R rats. I/R, ischemia-reperfusion; TMP, tetramethylpyrazine; KIM-1, kidney injury molecule 1; TNF-a, tumor necrosis factor-a; IL-, interleukin; SCr, serum creatinine; BUN, blood urea nitrogen.

Fig 5: The influences of geraniol (100 and 200 mg/kg) administration for 14 days prior to renal ischemia/perfusion (I/R) induced injury on the renal function assessment, including (a) creatinine, (b) BUN, (c) uric acid, (d) Kim-1, and (e) LDL. All values are expressed as mean ± SD. ¥ indicates statistically significant from the sham group, ₳ indicates statistically significant from the renal I/R group, and φ indicates statistically significant from renal I/R + geraniol 100 mg/kg group (p < 0.05) using one-way ANOVA followed by Tukey’s post hoc test.