Cytokine levels in plasma and peritoneal lavage fluid after intraperitoneal or intravenous anti-IL-17A antibody blockade.

<p>The mice underwent a sham procedure, CLP, or CLP with intraperitoneal or intravenous anti-IL-17A administration (<i>n</i> = 5 for each group). The levels of TNF-α and IL-6 in the blood (A, B) and peritoneal lavage fluid (C, D) were measured 12 h after surgery. **<i>P</i><0.01, *<i>P</i><0.05.</p

Superior effect of intraperitoneal vs intravenous IL-17A antibody administration.

<p>Intraperitoneal (A) or intravenous (B) anti-IL-17A antibody administration protected the mice from severe CLP. The CLP mice were intraperitoneally or intravenously given 50 µg of anti-IL-17A antibodies (<i>n</i> = 15), 50 µg of isotype control antibodies (<i>n</i> = 12), or 0.2 mL of PBS 3 h after severe CLP surgery. **<i>P</i><0.01: i.p. anti-IL-17A compared with the CLP group; ^##<i>P</i><0.01: i.p. anti-IL-17A compared with the i.p. isotype group; <i>P</i><0.05: i.v. anti-IL-17A compared with the CLP group.</p

Flow cytometric analysis of IL-17A–producing cells in peritoneal fluid after CLP and the percentage of neutrophil granulocytes in peritoneal fluid after intraperitoneal blockade with anti-IL-17A antibodies.

<p>(A) Representative flow cytometric dot plots. (B) Percentage of IL-17A+ -T cells in peritoneal lavage fluid after CLP or sham operation. (C) Percentage of neutrophil granulocytes in peritoneal fluid after blockade of anti-IL-17A in the four groups. The mice underwent a sham procedure, CLP, CLP with intraperitoneal anti-IL-17A or isotype administration (<i>n</i> = 6 for each group). Peritoneal lavage fluid was harvested 24 h after surgery. *<i>P</i><0.05.</p

Bacterial clearance after intraperitoneal anti-IL-17A antibody administration.

<p>The mice underwent a sham procedure, CLP, CLP with intraperitoneal anti-IL-17A, or isotype administration (n = 10 for each group). Blood and peritoneal lavage fluid were harvested 24 h after surgery. Mice that received intraperitoneal anti-IL-17A antibodies exhibited a deceased bacterial burden in the blood (A) and peritoneal cavity (B) compared with mice that received isotype antibodies or PBS. **P<0.01.</p

IL-17A concentration in blood and peritoneal lavage fluid after severe CLP.

<p>(A) IL-17A concentration in blood 3, 6, 12, and 24 h after the sham operation and CLP, which peaked at 12 h. (B) IL-17A concentration in peritoneal lavage fluid 3, 6, 12, and 24 h after the sham operation and CLP, which peaked at 6 h. **P<0.01: Compared with CLP 3 h group, IL-17A concentration elevated significantly. ##P<0.01 and #P<0.05: IL-17A concentration elevated significantly in CLP mice than sham group mice on the same time point.</p

Survival rate of septic mice after injection with baicalin.

<p>Baicalin improves survival of polymicrobial sepsis in CLP mice. In the bai group, mice were treated with intra-peritoneal baicalin (100 mg/kg) at 1 h, 6 h and 12 h after CLP. Normal saline of equal volume was administered in sham and CLP groups. Bai, baicalin group; CLP, CLP group. Survival analyses with log-rank test. ** <i>P</i><0.01.</p

Bacterial clearance in blood (A) and peritoneum (B) in septic mice after baicalin treatment.

<p>Baicalin enhanced bacterial clearance in blood (A) and peritoneum (B). Blood and peritoneal lavage fluid were harvested 24 h after CLP. Bai, baicalin group; CLP, CLP group. Data analyses with one-way ANOVA and Newman-Keuls. ** <i>P</i><0.01.</p

The number of neutrophils in peritoneum in septic mice after administration of baicalin.

<p>Baicalin reduced the number of neutrophils in peritoneum. Peritoneal lavage fluid were harvested 24 h after CLP. Bai, baicalin group; CLP, CLP group. Data analyses with one-way ANOVA and Newman-Keuls. * <i>P</i><0.05.</p

Histopathological changs of septic mice after baicalin treatment.

<p>The tissues were harvested 24 h after CLP for histopathologic examination using hematoxylin and eosin staining. Representative images from six animals per group were shown. Histopathological tests showed milder impairment in lung and liver after baicalin administration. A, After baicalin treatment, lung showed less neutrophil accumulation and alveolar destruction. B, Hepatocytes were protected and hepatic sinusoid was also preserved in baicalin treated mice. C, the severity of lung injury was scored as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035523#s2" target="_blank">Materials and methods</a>. D, the severity of liver injury was scored as above. ** <i>P</i><0.01 vs. sham. ## <i>P</i><0.01 vs. CLP. Bai, baicalin group; CLP, CLP group.</p