Andrographolide sulfate inhibited NF-κB activation and alleviated pneumonia induced by poly I:C in mice

Abstract

Pneumonia is a common illness that continues to be the major killer of remaining to be a significant source of morbidity and mortality in the patient population. Many microorganisms cause pneumonia, and now concern is turning to the importance of the cause the new therapies for viral pneumonia. In the current study, we report the effect of andrographolide sulfonate, a water-soluble form of andrographolide (trade name: Xi-Yan-Ping Injection), on poly I: C-induced pneumonia. Andrographolide sulfonate was administrated through intraperitoneal injection to mice with poly I: C-induced pneumonia. Recruitment of airway inflammatory cells, alteration of lung histological induced by Poly I: C were significantly ameliorated by andrographolide sulfonate. The protein levels of pro-inflammatory cytokines in bronchoalveolar fluid (BALF) and serum were reduced by andrographolide sulfonate treatment. The levels of MUC5AC and MUC5B in lung tissue were also suppressed. These results reveal that andrographolide sulfate remarkably alleviated pneumonia induced by poly I:C in mice. Moreover, andrographolide sulfonate markedly inhibited the activation of nuclear factor-κB (NF-κB). Taken together, we demonstrated that andrographolide sulfonate ameliorated poly I: C-induced pneumonia in mice, suggesting the possible use of andrographolide sulfonate for virus-induced pneumonia in clinical.

Keywords: Andrographolide; NF-κB; Pneumonia; Poly I:C; Sulfonate.

Fig. 1

Andrographolide sulfonate treatment ameliorated lung damage induced by poly I:C in mice. Lung tissue from each group were taken and fixed in 4% formalin and subjected to hematoxylin/eosin (H&E) staining (A, B) and Masson staining (C, D). Scale bar 100 μm. Andro-S: andrographolide sulfonate.

Fig. 2

Andrographolide sulfonate treatment prevented poly I:C-induced recruitment of inflammatory cells in lung tissue. Cells in BALF (bronchoalveolar lavage fluid) from each group were gathered and counted. Then they were stained with anti-CD3-APC, anti-CD11b-PE, anti-Gr1+-FITC and analyzed by FACS. Values were shown as the means ± SEM of 6 mice. ^∗P < 0.05, ^∗∗P < 0.01 vs. mice treated with poly I:C group. Andro-S: andrographolide sulfonate.

Fig. 3

Andrographolide sulfonate treatment suppressed poly I:C-induced cytokines expression. Cytokine levels in BALF supernatant (A) and serum (B) from each group were detected by ELISA. (C) RNA of lung tissue from the mice was extracted. The mRNA expression of cytokines was examined by real-time PCR. Values were shown as the means ± SEM of 6 mice. ^∗P < 0.05, ^∗∗P < 0.01 vs. mice treated with poly I:C group. Andro-S: andrographolide sulfonate.

Fig. 4

Andrographolide sulfonate suppressed mucins expression in lung of mice with poly I:C-induced pneumonia. (A) RNA of lung tissue from the mice in each group was extracted. The mRNA expression of MUC1, MUC5AC, MUC5B were examined by real-time PCR. Values were shown as the means ± SEM of 6 mice. (B) Paraffin-embedded lung tissue sections from each group were stained for MUC5AC. Scale bar: 100 μm ^∗P < 0.05, ^∗∗P < 0.01 vs. mice treated with poly I:C group. Andro-S: andrographolide sulfonate.

Fig. 5

Andrographolide sulfonate decreased activations of NF-κB in the lung of mice with poly I: C-induced pneumonia. Paraffin-embedded lung tissue sections from each group were stained for p-p65. Scale bar 100 μm. Andro-S: andrographolide sulfonate.