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. 2011:2011:604196.
doi: 10.1093/ecam/nep151. Epub 2011 Mar 10.

Proteomic Analysis of Anti-inflammatory Effects of a Kampo (Japanese Herbal) Medicine "Shoseiryuto (Xiao-Qing-Long-Tang)" on Airway Inflammation in a Mouse Model

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Proteomic Analysis of Anti-inflammatory Effects of a Kampo (Japanese Herbal) Medicine "Shoseiryuto (Xiao-Qing-Long-Tang)" on Airway Inflammation in a Mouse Model

Takayuki Nagai et al. Evid Based Complement Alternat Med. 2011.

Abstract

Effects of a Kampo (Japanese herbal) medicine "shoseiryuto (SST, xiao-qing-long-tang in Chinese)", which has been used for the treatment of allergic bronchial asthma clinically, were examined on ovalbumin (OVA)-sensitized allergic airway inflammation model (i.e., bronchial asthma) in a mouse. When SST was orally administered at 0.5 g kg(-1) day(-1) from day 1 to 6 after OVA inhalation, SST reduced the inflammation in lung tissue, the number of eosinophils and the OVA-specific immunoglobulin E (IgE) antibody titer in bronchoalveolar lavage (BAL) fluids at 7 days after the OVA inhalation. SST also reduced the airway hyperreactivity at 6 days after the OVA inhalation. Proteomic analysis with the agarose two-dimensional electrophoresis showed that the expression of spectrin α2 was reduced in the lung tissue of OVA-sensitized mice and SST recovered the expression. Western blot and immunohistochemical analyses of lung tissue also confirmed this result. When prednisolone was orally administered at 3 mg kg(-1) day(-1) from day 1 to 6 after OVA inhalation, the inflammation in lung tissue, the number of eosinophils in BAL fluids and airway hyperreactivity were reduced in the OVA-sensitized mice. However, prednisolone did not reduce the OVA-specific IgE antibody titer in BAL fluids and did not recover the expression of spectrin α2 in lung tissue. These results suggest that at least a part of action mechanism of SST against OVA-sensitized allergic airway inflammation in a mouse model is different from that of prednisolone.

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Figures

Figure 1
Figure 1
Chemical profile of the hot water extract from shoseiryuto as analyzed by 3D HPLC.
Figure 2
Figure 2
Effects of SST and prednisolone on lung tissue eosinophilia, hypertrophied smooth muscle and mucus secretion in OVA-sensitized mice were measured. Non-sensitized BALB/c mice were treated p.o. with water (a and e), and OVA-sensitized mice were treated p.o. with water (b and f), SST (0.5 g kg−1) (c and g) or prednisolone (3 mg kg−1) (d and h) five times from day 1 to 6 after OVA exposure. Lung tissues were removed at 7 days after OVA exposure and stained with hematoxylin and eosin (a–d) or periodic acid-Schiff (e–h) as described in “Methods” section. Magnification ×100.
Figure 3
Figure 3
Effects of SST and prednisolone on number of eosinophils in blood and BAL fluid of OVA-sensitized mice were measured. OVA-sensitized mice were treated with SST, prednisolone (PRE) or water as described in legend of Figure 2. Number of eosinophils in blood (a) and BAL fluid (b) were counted on 7 days after the OVA exposure. Each column represents the mean  ±  SEM of five to seven mice per group. Statistical analysis was conducted using Tukey's test.
Figure 4
Figure 4
Effects of SST and prednisolone on differential cell number in blood and BAL fluid of OVAsensitized mice were measured. OVA-sensitized mice were treated with SST, prednisolone (PRE) or water as described in legend of Figure 2. Differential cell number in blood (a) and BAL fluid (b) were counted on 7 days after the OVA exposure. Each column represents the mean  ±  SEM of five to seven mice per group. # P < .0001 versus control; *P < .001 and **P < .0001 versus water/OVA-sensitized group (Tukey's test). mac (macrophage), lym (lymphocyte), neu (neutrophil), eo (eosinophil), baso (basophil), mono (monocyte).
Figure 5
Figure 5
Effects of SST and prednisolone on OVA-specific IgE antibody titers in OVA-sensitized mice were measured. OVA-sensitized mice were treated with SST, prednisolone (PRE) or water as described in legend of Figure 2. OVA-specific IgE antibody titers in plasma (a) and BAL fluid (b) were determined at 7 days after OVA exposure. Each column represents the mean  ±  SEM of five to six mice per group. Statistical analysis was conducted using Tukey's test.
Figure 6
Figure 6
Effects of SST and prednisolone on airway hyperreactivity (AHR) in OVA-sensitized mice were measured with plethysmography. Non-sensitized mice were treated with water (closed circle), and OVA-sensitized mice were treated with SST (closed inverted triangle), prednisolone (open inverted triangle) or water (open circle) as described in legend of Figure 2. AHR to aerosolized methacholine (MeCh) was measured in unrestrained conscious mice at 1 h after the final treatment of drug. Mice were placed into the main chamber and were nebulized with PBS or MeCh (3.125, 6.25, 12.5, 25, 50 mg ml−1) for 3 min. Recording of Penh was taken for 5 min after each MeCh nebulization. Each bar represents the mean  ±  SEM of four mice per group. # P < .05, ## P < .01 and ### P < .001 versus water/non-sensitized group (Fisher's PLSD test). *P < .05, **P < .01 and ***P < .001 versus water/OVA-sensitized group (Dunnett's test).
Figure 7
Figure 7
Effect of SST on protein expression in lung tissues of OVA-sentitized mice was measured by agarose 2D electrophoresis. OVA-sensitized or non-sensitized mouse was treated with SST or water as described in legend of Figure 2. Lung tissue was obtained at 7 day after the OVA exposure. Agarose 2D electrophoresis patterns of non-sensitized/water-treated mouse (a), OVA-sensitized/water-treated mouse (b), non-sensitized/SST-treated mouse (c) and OVA-sensitized/SST-treated mouse (d), and results of statistical analysis of spectrin α2 expression (e). Each column represents the mean  ±  SEM of four mice per group. Statistical analysis was conducted using Tukey's test.
Figure 8
Figure 8
Effects of SST and prednisolone on spectrin α2 expression level in lung tissue of OVA-sensitized mice were measured by western blot analysis. OVA-sensitized mice were treated with SST, prednisolone (PRE) or water as described in legend of Figure 2. Lung tissue was obtained at 7 day after the OVA exposure. Western blot with β-actin was performed to verify that equivalent amounts of proteins were loaded in each lane. Western blotting patterns of spectrin α2 and β-actin (a), and results of statistical analysis of spectrin α2/β-actin ratio (b). Each column represents the mean  ±  SEM of five to seven mice per group. Statistical analysis was conducted using Tukey's test.
Figure 9
Figure 9
Effects of SST and prednisolone on spectrin a2 expression in lung of OVA-sensitized mice were measured by immunohistochemical staining. The spectrin a2 was stained with antispectrin a2 antibody. OVA-sensitized mice were treated with SST, prednisolone or water as described in legend of Figure 2. Lung tissue was obtained at 7 day after the OVA exposure. (a) Non-sensitized/water-treated mouse, (b) OVA-sensitized/watertreated mouse, (c) OVA-sensitized/SST-treated mouse and (d) OVA-sensitized/prednisolone-treated mouse. Magnification ×200.
Figure 10
Figure 10
Hypothetical diagram of therapeutic effects of SST on airway inflammation.

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