Fig 2: Effect of iNOS activity on the survival following endotoxic-induced lung injury. (A) Time-to-survival curves following intrapleural instillation of LPS in neonatal and adult rats. Compared with adults, mortality in neonates increased up to 10 h following pulmonary sepsis. Pre-treatment with a selective iNOS inhibitor, 1400w, significantly improved the survival time in neonates. *P=0.02 vs. neonates, assessed using the log-rank test. n=9 in adults+LPS, n=14 in neonates+LPS and n=14 in neonates+LPS+1400w groups. (B) Generation of NO following induction of pulmonary sepsis. Lung tissue concentrations of NOx were assessed using a Griess reaction assay kit. NOx levels were significantly decreased in lung homogenates of neonates treated with 1400w. *P=0.043 vs. adults; #P<0.001 vs. adults, assessed via Kruskal-Wallis. n=9 in neonates+LPS, n=9 in neonates+LPS+1400w and n=8 in adults+LPS groups. (C) Formation of 3-NT following induction of pulmonary sepsis. Lung tissue concentrations of 3-NT were measured using a 3-NT ELISA kit. *P=0.003 vs. adults; #P<0.001 vs. adults, assessed via Kruskal-Wallis. n=11 in neonates+LPS, n=11 in neonates+LPS+1400w and n=8 in adults+LPS groups. Dotted lines (B and C) indicate the tissue concentrations in the neonatal lungs at basal unstimulated condition. LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; NOx, nitrite and nitrate; 3-NT, 3-nitrotyrosine.

Fig 3: Effects of erinacine C (EC) on LPS-induced iNOS expression in BV2 microglial cells. Cells (1 × 105 cells/mL) were pretreated with different concentrations (0.1–2.5 µM) of EC for 1 h, after which the cells were treated with LPS (500 ng/mL) for 24 h. iNOS protein expression was determined by western blot analyses. Values (means ± SD, n = 3) not sharing a common lower case letter are significantly different (p < 0.05). The western blot image is provided as a representative result of multiple experiments.

Fig 4: Effect of SST on LPS-induced expression of Cox-2, iNOS and the activation of NF-?B p65. l. (A and C) representative western blot gels showing Cox-2, iNOS, NF-?B p65 and NF-?B p-p65 levels. Cox-2, iNOS and NF-?B p-p65 are represented by the bands at 21, 130 and 65 kDa, respectively. ß-actin was used as an internal control. (B and D) Quantification of A and C by gray value analysis. Values are presented as the mean ± standard error of the mean of eight animals per group. *P<0.05, **P<0.01, ***P<0.001 compared with phosphate-buffered saline-injected SN. LPS, lipopolysaccharide; SST, somatostatin; SN, substantia nigra; COX, cyclooxygenase; iNOS, inducible nitric oxide synthase; NF-?B, nuclear factor kappa B.

Fig 5: Effects of Hirudo lyophilized powder on the inflammatory cytokine levels and macrophage infiltration in rat renal tissues. The SL, SM, and SH group rats were dosed by oral gavage once daily for 16 weeks with Hirudo lyophilized powder. The C and DM group rats were administered the same volume of vehicle used to prepare the test medications. (a) The mRNA levels of IL-1ß, MCP-1, and TNF-a in each group rats. Data are presented as means ± SD (n = 3). (b, c) Immunofluorescence staining for CD68 and iNOS in the renal tissues of each group rats; scale bar, 100 m. (d, e) The fluorescence intensity of CD68 and iNOS in each group rats. Data are presented as means ± SD. **P < 0.01 vs. C group; #P < 0.05 and ##P < 0.01 vs. DM group.