Inhibitory Activities of Rare Ginsenoside Rg4 on Cecal Ligation and Puncture-Induced Sepsis

Figures

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  Figure 1. Protection of mice from the morbidity and mortality of sepsis by Rg4 after the CLP procedure. (A) Survival rates after the induction of different (mid or high) severity grades of sepsis by CLP. (B,C) Each group consisted of 10 mice and received intravenous injections of vehicle, MA (0.7 mg/kg) or Rg4 (5, 10, or 15 mg/kg) in 0.5% DMSO at 24 h after mid- (B) or high-grade (C) CLP. Survival rates of challenged mice were monitored for 10 days. (D) Body weight changes were measured during the period in Figure (B). All values are presented as the mean ± SD of three separate experiments. * p < 0.01 vs. CLP group for Rg4 or MA in figure (D).

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  Figure 2. Inhibition of inflammatory cytokines and renal and hepatic inflammation by Rg4 in CLP. Each group consisted of 5 mice, which received intravenous injections of vehicle, MA (0.7 mg/kg) or Rg4 (5, 10, or 15 mg/kg) in 0.5% DMSO at 24 h after high-grade CLP. At 72 h after CLP, the kidneys or livers were homogenized and used for the determination of TNF-α (A) or IL-1β (B) in kidneys (A,B) or livers (C). All values are presented as the mean ± SD of three separate experiments. # p < 0.01 vs. control group or * p < 0.01 vs. CLP group. (D) Kidney tissues were stained with H&E. Photomicrographs of kidney tissues at 72 h after CLP are shown (original magnification, ×200); the images are representative of three independent experiments. Scale bars: 160 μm.

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  Figure 3. Inhibition of HMGB1-induced TLR/NF-κB expression by Rg4 in vitro. (A) HUVECs were activated with HMGB1 (1 μg/mL) treatment for 6 h, followed by treatment with various concentrations of Rg4 for another 6 h. Cell lysates were subjected to Western blot analysis to determine the protein levels of TLR2, TLR4, and NF-κB. Representative Western blots (left) and relative densitometric quantification (right) of each gene. (B) Supernatants from (A) were used for TNF-α ELISA. All values are presented as the mean ± SD of three separate experiments. D = 0.5%, DMSO was the vehicle control and H = HMGB1. # p < 0.01 vs. DMSO or * p < 0.01 vs. HMGB1 group.

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  Figure 4. Increased cell survival through induction of PI3K/AKT signaling by Rg4 in HMGB1-mediated sepsis in vitro. HUVECs were activated with HMGB1 (1 μg/mL) for 6 h and then treated with various concentrations of Rg4 for another 6 h. (A) The MTT assay was performed to assess cell viability. Cell lysates were subjected to Western blot analysis to estimate protein levels of PI3K, p-AKT, and total AKT (B) or to ELISA for phosphor-PI3K (C). Representative Western blots (left) and relative densitometric quantification (right) of each gene. All values are presented as the mean ± SD of three separate experiments. D = 0.5%, DMSO was the vehicle control and H = HMGB1. # p < 0.01 vs. DMSO or * p < 0.01 vs. HMGB1 group.