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, 2019, 4827342
eCollection

Shuang-Huang-Lian Attenuates Airway Hyperresponsiveness and Inflammation in a Shrimp Protein-Induced Murine Asthma Model

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Shuang-Huang-Lian Attenuates Airway Hyperresponsiveness and Inflammation in a Shrimp Protein-Induced Murine Asthma Model

Yuan Gao et al. Evid Based Complement Alternat Med.

Abstract

Shuang-Huang-Lian (SHL), an herbal formula of traditional Chinese medicine, is clinically used for bronchial asthma treatment. Our previous study found that SHL prevented basophil activation to suppress Th2 immunity and stabilized mast cells through activating its mitochondrial calcium uniporter. Sporadic clinical reports that SHL was used for the treatment of bronchial asthma can be found. Thus, in this study, we systematically investigated the effects of SHL on asthmatic responses using a shrimp protein (SP)- induced mouse model. SHL significantly inhibited airway inspiratory and expiratory resistance, and histological studies suggested it reduced thickness of airway smooth muscle and infiltration of inflammation cells. It also could alleviate eosinophilic airway inflammation (EAI), including reducing the number of eosinophils and decreasing eotaxin and eosinophil peroxidase levels in the bronchoalveolar lavage fluid (BALF). Further studies indicated that SHL suppressed SP-elevated mouse mast cell protease-1 and IgE levels, prevented Th2 differentiation in mediastinal lymph nodes, and lowered Th2 cytokine (e.g., IL-4, IL-5, and IL-13) production in BALF. In conclusion, SHL attenuates airway hyperresponsiveness and EAI mainly via the inhibition of mast cell activation and Th2 immunity, which may help to elucidate the underlying mechanism of SHL on asthma treatment and support its clinical use.

Figures

Figure 1
Figure 1
Electrophoretogram of SP by SDS-PAGE.
Figure 2
Figure 2
Protocol for the induction of murine asthma model and SHL treatment.
Figure 3
Figure 3
SHL prevents SP-induced AHR in the murine asthma model. (a-b) SHL prevents SP-induced AHR by comparing RL (a) and Re (b) values with NS mice. 24 h after the last nebulization of SP, the mice were treated (i.v.) with Mch at the indicated doses and RL and Re values were recorded by AniRes 2005 mouse lung function analysis system. (c) Representative histological images of AR in lung tissue. Values were expressed as mean ± SD (n = 8). ##P < 0.01 versus NS; P < 0.05 and ∗∗P < 0.01 versus SP.
Figure 4
Figure 4
Effect of SHL on SP-induced EAI. (a) Representative images of eosinophil infiltration in BALF. The mice were sacrificed and the BALF was collected. Cytospin preparations were made for the leukocytes differential cell count by Wright's staining. (b) The proportion of eosinophils in leukocytes in BALF. (c-d) SHL reduced eotaxin and EPO levels in BALF. Values were expressed as mean ± SD (n = 8). ##P < 0.01 versus NS; P < 0.05 and ∗∗P < 0.01 versus SP.
Figure 5
Figure 5
Effects of SHL on SP-elevated IgE and mMCP-1 levels. (a-b) SHL reduced tIgE and sIgE in serum. (c-d) SHL decreased tIgE and mMCP-1 levels in BALF. Values were expressed as mean ± SD (n = 8). ##P < 0.01 versus NS; P < 0.05 and ∗∗P < 0.01 versus SP.
Figure 6
Figure 6
Effect of SHL on Th2 cytokines in BALF and their production in MLNs cells. (a) SHL decreased IL-4, IL-5, and IL-13 levels in BALF. (b) SHL decreased IL-4, IL-5, and IL-13 production in MLNs cells. Three days after the last inhalation of SP, mice were sacrificed and the MLNs were isolated. Single cell suspensions were aseptically prepared and seeded in a round bottom 96-well plate (2 × 106 cells/well) and then stimulated with SP (10 μg/mL) at 37°C for 72 h. The levels of IL-4, IL-5, and IL-13 in the culture medium were measured using the ELISA kits. Values were expressed as mean ± SD (n = 8). ##P < 0.01 versus NS; P < 0.05 and ∗∗P < 0.01 versus SP.

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