Inhibition of the Interactions Between Eosinophil Cationic Protein
and Airway Epithelial Cells by Traditional Chinese Herbs
By Hao-Teng Chang, Louis J Tseng, Ta-Jen Hung, Blacky T Kao, Wei-Yong Lin,
Tan-chi Fan, Margaret Dah-Tsyr Chang, Tun-Wen Pai
Background: The eosinophil cationic protein (ECP) is cytotoxic to bacteria, viruses, parasites and mammalian cells. Cells are damaged via processes of pore formation, permeability alteration and membrane leaking. Some clinical studies indicate that ECP gathers in the bronchial tract of asthma sufferers, damages bronchial and airway epithelial cells, and leads to in breathing tract inflammation; therefore, prevention of the cytotoxicity caused by ECP may serve as an approach to treat airway inflammation. To achieve the purpose, reduction of the ECP-cell interactions is rational. In this work, the Chinese herbal combinative network was generated to predict and identify the functional herbs from the pools of prescriptions. It was useful to select the node herbs and to demonstrate the relative binding ability between ECP and Beas-2B cells with or without herb treatments.
Results: Eighty three Chinese herbs and prescriptions were tested and five effective herbs and six prescription candidates were selected. On the basis of effective single-herbal drugs and prescriptions, a combinative network was generated. We found that a single herb, Gan-cao, served as a node connecting five prescriptions. In addition, Sheng-di-huang, Dang-guei and Mu-tong also appeared in five, four and three kinds of prescriptions, respectively. The extracts of these three herbs indeed effectively inhibited the interactions between ECP and Beas-2B cells. According to the Chinese herbal combinative network, eight of the effective herbal extracts showed inhibitory effects for ECP internalizing into Beas-2B cells. The major components of Gan-cao and Sheng-di-huang, glycyrrhizic acid and verbascose, respectively, reduced the binding affinity between ECP and cells effectively.
Conclusions: Since these Chinese herbs reduced the binding affinity between ECP and cells and inhibited subsequent ECP entrance into cells, they were potential for mitigating the airway inflammation symptoms. Additionally, we mentioned a new concept to study the Chinese herbs using combinative network in the field of systems biology. The functional single herbs could be identified from the set of prescriptions.
- Results and Discussions
- The pre-screening of Chinese herbs: two herbal sets containing 90 and 83 kinds of herbs or prescriptions were selected and purchased from Sun Ten Pharmaceutical Co. Ltd. and another pharmaceutical Co. Ltd., separately.
- Gan-cao, Sheng-si-huang, Dan-guei and Mu-tong were potential for inhibiting the ECP-cell interactions
- The inhibition of ECP translocating into Beas-2B cells
- The major components of herbs inhibited the interaction of ECP and Beas-2B cells
- Materials and Methods
- The flowchart
- Antibodies and reagents
- Cells and cell culture
- Purification of recombinant proteins
- Preparation of Chinese herbs
- Cell-based ELISA
- Drug treatment
- Generation of combinative network
- Immuno-fluorescence staining and confocal microscopy
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