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《Traditional Chinese Drug Research and Clinical Pharmacology》 2019-12
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Huatan Jiangqi Capsule Containing Serum Up-regulates MRP1 via Notch Pathway in Human Bronchial Epithelial Cells

LI Nini;JIANG Chengjun;GUO Yan;WANG Dianlei;Graduate School, Anhui University of ChineseMedicine;Laboratory of Drug Metabolism and Pharmacokinetics, AnhuiUniversity of Chinese Medicine;  
Objective The aim of this study was to investigate the effect and mechanism of Huatan Jiangqi capsule containing serum on the expression and function of MRP1 based on the Notch signaling pathway through in vitro experiments. Methods According to the method of serum pharmacology,the drug containing serum and blank serum were prepared, and then 5%, 10%, 15% and 20% drug containing serum were used to intervene with human bronchial epithelial(16 HBE)cells. CCK-8 kit was used to detect the proliferation of 16 HBE cells for the optimal concentration. Cells were divided into blank serum control group,Huatan Jiangqi capsule containing serum group,cigarette smoke extract(CSE)+ blank serum group, CSE + drug containing serum group, DAPT + blank serum group,DAPT+Huatan Jiangqi capsule containing serum group. Western Blot was used to detect protein expression levels of Notch1,Hes1 and MRP1. Using fluorescein 5-carboxydiacetic acid(5-CFDA)as substrate,the change of intracellular fluorescence intensity in 16 HBE cells was detected by flow cytometry, which can reflect the efflux function of MRP1. The NICD nuclear translocation activity was detected by immunofluorescence. Results The optimal concentration of drug containing serum was 15%. Compared with the blank control group,the expression levels of Notch1,Hes1 and MRP1 protein were significantly up-regulated in Huatan Jiangqi capsule containing serum group(P 0.05,P 0.01),and the increase was concentration-dependent. The efflux capacity of MRP1 transporter was significantly improved(P 0.01)and the fluorescence intensity of NICD in the nucleus was enhanced. While the protein expression levels in CSE+ blank serum group were significantly reduced(P 0.01). Compared with the CSE+blank serum group, protein expression levels of Notch1, Hes1 and MRP1 were all increased in the CSE + drug containing serum group(P 0.05,P 0.01). Compared with the Huatan Jiangqi capsule containing serum groups,the expression levels of Notch1,Hes1 and MRP1 protein in DAPT+Huatan Jiangqi capsule containing serum group were all decreased(P 0.05, P 0.01), MRP1 efflux capacity was significantly decreased(P 0.01), NICD fluorescence intensity in cytoplasmic and nuclei were all weakened. Conclusion Huatan Jiangqi capsule containing serum can up-regulate the expression and efflux function of MRP1 in 16 HBE cells through Notch pathway,which may be one of the mechanisms of its treatment of chronic obstructive pulmonary disease(COPD).
【Fund】: 国家自然科学基金项目(81473536)
【CateGory Index】: R285.5
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