Anti-inflammatory activity of Alchornea triplinervia ethyl acetate fraction: Inhibition of H₂O₂, NO and TNF-α

Figures & data

Table 1.  Major constituents of A. triplinervia ethyl acetate fraction (AtF). The symbol (×) indicates the presence and (–) the absence of the classes of compounds.

Classes of compounds	AtF
Gallic acid derivatives	×
Glycoside flavonoids	×
Phenolic acids	×
Alkaloids	–
Tannins	–
Saccharides	–
Lipids	–
Proteins	–

Figure 1.  Effects of A. triplinervia ethyl acetate fraction (AtF) on the viability of peritoneal macrophages in the presence of PMA and LPS. For the test using PMA, PEC (2 × 10⁶) was utilized and the adherent cells were incubated for 1h with the fraction and PMA (0.2 μM). For the test using LPS, PEC (5 × 10⁶) was utilized and the adherent cells were incubated with the fraction and LPS (1 μg/mL) for 24 h. Cells in culture medium (control) correspond to 100% of viability. The cell viability was determined by MTT assay as described previously. Results are the means ± SD of five separate experiments. One-way ANOVA with Dunnett’s post test was performed.*p <0.01 versus control; **p <0.05 versus control.

Table 2.  Inhibitory effects of A. triplinervia ethyl acetate fraction (AtF) on H₂O₂, NO and TNF-α production.

Inhibition (%)
	AtF concentrations
	15.62 µg/mL	62.5 µg/mL
H2O2 production	72.25 ± 4.68	69.64 ± 4.21
NO production	47.8 ± 8.96	76.77 ± 8.11
TNF-α production	16.41 ± 6.38	21.02 ± 8.67

Figure 2.  Effects of A. triplinervia ethyl acetate fraction (AtF) on H₂O₂ production in peritoneal macrophages. PEC at 2 × 10⁶cells/mL was used and complete buffer was added to the adherent cells. The cells were exposed to the fraction and PMA 0.2 μM. Cells incubated only with PMA were used as a positive control and cells in complete buffer as a negative control (C). Results are the means ± SD of five separate experiments. One-way ANOVA with Dunnett’s post test was performed. *p <0.01 versus PMA control.

Figure 3.  Effects of A. triplinervia ethyl acetate fraction (AtF) on NO production in peritoneal macrophages. PEC at 5 × 10⁶ cells/mL was used and the adherent cells were incubated for 24 h with the fraction and LPS (1 μg/mL). Cell-free supernatant was mixed with Griess reagent. Cells incubated only with LPS were used as a positive control and cells in culture medium (RPMI-1640) as a negative control (C). Results are the means ± SD of five separate experiments. One-way ANOVA with Dunnett’s post test was performed. *p <0.01 versus LPS control.

Figure 4.  Effects of A. triplinervia ethyl acetate fraction (AtF) on TNF-α production in peritoneal macrophages. For the cytokine immunoassay, PEC at 5 × 10⁶/mL was used. Adherent cells were incubated for 24 h with the fraction and LPS (1 μg/mL). Cells incubated only with LPS were used as a positive control and cells in culture medium (RPMI-1640) as a negative control (C). Results are the means ± SD of five separate experiments. One-way ANOVA with Dunnett’s post test was performed. *p<0.01 versus LPS control.