Fig 2: Fag decreases the plasma levels of (A) LPS, (B) IL-1ß, (C) IL-6 and (D) TNF-a in TNBS-induced colitis mice. n=6. #P<0.01 vs. control group; *P<0.05 and **P<0.01, vs. TNBS group. Fag, Fagopyrum cymosum (Trev.) Meisn; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor; TNBS, 2,4,6-trinitrobenzenesulfonic acid; SASP, salicylazosulfapyridine.

Fig 3: Anti-IFP35 protected C57BL/6J mice from lethal influenza virus infection(A) Schematic of the experimental protocol using anti-IFP35 against influenza infection.(B–D) Body weight loss (percent) (B), clinical score (C), and survival rate (percent) (D) of anti-IFP35- or mIgG-treated mice under 2 × 106 PFU PR8 virus infection; n = 12.(E) H&E staining of the lung tissue. Scale bars, 100 µm. Multifocals with collagen fibrin exudation are indicated by a red arrow. Inflammatory cells are indicated by a black arrow.(F) The pathological score of the lung tissue in anti-IFP35-treated mice decreased.(G) Serum TNF in anti-IFP35- or mIgG-treated mice infected with 2LD50 PR8 virus.(H) PR8 viral genome RNA (nucleoprotein, NP) in the lung tissue of C57BL/6J mice; n = 3.Data in (F)–(H) are mean values ± SEM. Significance in (B) and (C) was calculated with two-way ANOVA with Sidak’s post-test. Significance in (D) was determined using log rank test (*p < 0.05, **p < 0.01). Significance in (F)–(H) was assessed by Mann-Whitney U test (*p < 0.05, **p < 0.01, ***p < 0.001; ns, no significance).

Fig 4: Malva parviflora hydroalcoholic extract attenuates microglia pro-inflammatory M1 phenotype in the cortex of 5XFAD mice. Total RNA was isolated from the cortex of Wt or 5XFAD mice fed with either normal diet (ND) or high-fat diet (HFD) non-treated (Vehicle) or treated with MpHE (M. parviflora) for 8 months. a The transcript levels of CD86 (marker of M1 state) were determined by RT-qPCR as described in the “Methods” section. Data are shown as mean ± SEM, n = 3 animals per group. Statistical analysis was performed by three-way ANOVA followed by Tukey’s multiple comparisons test. This analysis revealed a significant effect for the genotype F(1,16) = 42.45, p < 0.001; for the diet F(1,16) = 0.4022, p = 0.53; for the M. parviflora treatment F(1,16) = 29.79, p < 0.001; for the genotype and diet interaction F(1,16) = 0.04041, p = 0.84; for the M. parviflora treatment and diet interaction F(1,16) = 0.2594 p = 0.62; for the genotype and M. parviflora treatment interaction F(1,16) = 20.67 p < 0.001; for the genotype, M. parviflora treatment and diet interaction F(1,16) = 2.037, p = 0.17. b TNF (marker of M1 state) mRNA levels. Data are shown as mean ± SEM, n = 3 animals per group. Statistical analysis was performed by three-way ANOVA followed by Tukey’s multiple comparisons test. This analysis revealed a significant effect for the genotype F(1,16) = 25.65 p < 0.001; for the diet F(1,16) = 5.758, p = 0.03; for the M. parviflora treatment F(1,16) = 32.4, p < 0.001; for the genotype and diet interaction F(1,16) = 4.955, p = 0.04; for the M. parviflora treatment and diet interaction F(1,16) = 2.259 p = 0.15; for the genotype and M. parviflora treatment interaction F(1,16) = 26.77 p < 0.001; for the genotype, M. parviflora treatment and diet interaction F(1,16) = 2.189, p = 0.16. c Mgl1 (marker of M2 state) mRNA levels. Data are shown as mean ± SEM, n = 3 animals per group. Statistical analysis was performed by three-way ANOVA followed by Tukey’s multiple comparisons test, and d TREM-2 mRNA levels. Data are shown as mean ± SEM, n = 3 animals per group. Statistical analysis was performed by three-way ANOVA followed by Tukey’s multiple comparisons test. Microglia from 8-month-old Wt or 5XFAD mice were unstimulated or stimulated with LPS (100 ng/mL) in the presence or absence of MpHE (M. parviflora; 1 mg/mL) for 24 h. Control cells were treated with PBS (Ctrl) or MpHE alone (M. parviflora). Supernatants were used to determine TNF and IL6 levels by ELISA as described in the “Methods” section. e TNF levels. Data are shown as mean ± SEM, n = 3 animals per group. Statistical analysis was performed by three-way ANOVA followed by Tukey’s multiple comparisons test. This analysis revealed a significant effect for the genotype F(1,16) = 7.878, p = 0.0127; for the LPS treatment F(1,16) = 17.74, p = 0.0007; for the M. parviflora treatment F(1,16) = 66.30, p < 0.0001; for the genotype and LPS treatment interaction F(1,16) = 6.105, p = 0.0251. f IL6 levels. Data are shown as mean ± SEM, n = 3 animals per group. Statistical analysis was performed by three-way ANOVA followed by Tukey’s multiple comparisons test. This analysis revealed a significant effect for the genotype F(1,16) = 25.76, p = 0.0001; for the LPS treatment F(1,16) = 19.86, p = 0.0004; for the M. parviflora treatment F(1,16) = 309.3, p < 0.0001; for the genotype and LPS treatment interaction F(1,16) = 20.71, p = 0.0003

Fig 5: Two milligrams of MSU crystals in 1 mL of PBS were injected into the peritoneum of mice in the presence or absence of BPI (50 µg/mL). In a group of mice, crystals were co-injected with a composition of BPI 50 µg/mL (1.5 µg/g body weight) and HA (0.02%). Controls received the same volume of sterile PBS alone. A group of mice was injected with crystals in the presence of HA (0.02%). Mice were sacrificed after 3 h, and (A) the number of leucocytes (white blood cells (WBC)) and (B) the percentage of polymorphonuclear cells (PMN) accumulated in peritoneal fluids were determined. Supernatants from the peritoneal wash fluids were analyzed for (C) IL-1ß, (D) CXCL1, (E) IL-6, (F) TNF-a, and (G) VEGF. Results are presented as the mean ± SD of 14 mice per group. *** p < 0.001 vs. MSU group, # p < 0.05 vs. MSU + BPI group, ## p < 0.01 vs. MSU + BPI group, ### p < 0.001 vs. MSU + BPI group, $ p < 0.05 vs. MSU + BPI + HA group, $$$ p < 0.001 vs. MSU + BPI + HA group. BPI: bactericidal/permeability-increasing protein, CXCL1: chemokine (C–X–C motif) ligand 1, HA: hyaluronic acid, IL: interleukin, PBS: phosphate-buffered saline, MSU: monosodium urate crystals, PMN: polymorphonuclear cells, TNF: tumor necrosis factor, VEGF: vascular endothelial growth factor, WBC: white blood cells.