1.A high throughput screening model for P-gp functional inhibitors
Tingyuan LANG ; Jufang YAN ; Changhua HU
Chinese Pharmacological Bulletin 2003;0(10):-
Aim To establish a cell model for high throughput screening and search for novel inhibitors of P-gp.Methods A fluorescent cell-based assay was developed for application in high throughput screening.A library of 480 traditional Chinese medicine samples was screened. Results The value of Z′ was 0.72. Screening of a library of 480 traditional Chinese medicine identified 2 positive samples.Conclusions This HTS model for screening P-gp inhibitors was stable and sensitive.The 2 positive samples had good P-gp inhibitory activities.
2.Effects of Nicorandil on the Proliferation ,Migration Ability and Hippo/YAP Signaling Pathway of Pulmonary Artery Smooth Muscle Cells
Fengnan CHEN ; Tingyuan LANG ; Changyu WU ; Xiaojun YU ; Xinmu SHI ; Kai SHEN ; Haiyan YANG
China Pharmacy 2020;31(22):2736-2740
OBJECTIVE:To evaluate the effects of nicorand il on the proliferation ,migration ability and Hippo/YAP signaling pathway of pulmonary artery smooth muscle cells (PASMCs). METHODS :Human primary PASMCs were divided into normal control group ,model group ,nicorandil low ,medium and high concentration groups (50,100,200 μmol/L),with 3 holes in each group. In addition to the normal control group ,the rest of the cells were inoculated on the gel coated medium to simulate the pulmonary hypertension environment ,so as to establish AS cell model. Then ,each drug group was added with corresponding drugs,and the normal control group and model group were added with the same volume of normal saline ,and cultured for 48 h. CCK-8 assay and Transwell assay were used for the examination of cell proliferation (by light density )and migration ability , respectively. mRNA expression of YAP target factors (CTGF and AREG )were examined by qRT-PCR. Western blotting assay was used to detect the protein expression of CTGF and AREG. RESULTS :Compared with normal control group ,light density of cells was increased significantly in model group ;the number of migration cells per field of view increased significantly ;mRNA and protein expression of CTGF and AREG were significantly increased (P<0.01). Compared with model group ,light density ,the number of migration cells per field of view ,mRNA and protein expression of CTGF and AREG in nicorandil low ,medium and high concentration groups were decreased significantly , in concentration-dependent manner (P<0.05 or P<0.01). CONCLUSIONS:Nicorandil can inhibit the proliferation and migration of PASMCs in AS model ,the mechanism of which cstc2019jscx-msxmX0174) may be associated with the Hippo/YAP signaling pathway.