Journal: International Journal of Molecular Sciences

Article Title: The Effect of Cholesterol Efflux on Endothelial Dysfunction Caused by Oxidative Stress

doi: 10.3390/ijms24065939

Figure Lengend Snippet: H 2 O 2 can promote cholesterol accumulation by oxidative stress. ( A , B ) The protein levels of LXRα, LXRβ, HMGCR, ABCA1, and ABCG1 were measured by a Western blot analysis. ( C ) The accumulation of ER cholesterol after filipin staining was visualized under a fluorescence microscope (magnification 5×). n = 3, * p < 0.05, and ** p < 0.001 vs. the C group (treated by PBS). C, control group; H 2 O 2 , cells treated with H 2 O 2 (500 μmol/L) for 24 h. The scale of the pictures was 100 μm.

Article Snippet: All media were filled with 10% FBS, and all cell cultures were carried out in an environment at 37 °C containing 5% CO 2 , H 2 O 2 (500 μmol/L, Guangdong Hengjian Pharmaceutical Co., Ltd., Jiangmen, China), LXR-623 (5 μmol/L, HY10629, MedchemExpress, Shanghai, China), cholesterol (100 μmol/L, C8280, Solaibio, Beijing, China), tunicamycin (10 μmol/L, MB5419, Meilunbio, Dalian, China), and salinomycin (5 μmol/L, HY-15597, MedchemExpress, Shanghai, China).

Techniques: Western Blot, Staining, Fluorescence, Microscopy, Control

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Cholesterol Efflux on Endothelial Dysfunction Caused by Oxidative Stress

doi: 10.3390/ijms24065939

Figure Lengend Snippet: LXR-623 can promote cholesterol efflux, and adding cholesterol extracellularly can induce cholesterol accumulation. ( A , C ) The expressions of ABCA1 and ABCG1 were detected by Western blot. ( B , D ) Filipin staining detected the intracellular cholesterol of HUVECs (magnification 5×). n = 3, * p < 0.05, and ** p < 0.001 vs. the C group; # p < 0.05, and ## p < 0.001 vs. the H 2 O 2 + LXR-623 group; # p < 0.05, and ## p < 0.001 vs. the H 2 O 2 + CHOL group. The C (control) group of LXR-623 and cholesterol were treated by DMSO and chloroform, respectively; H 2 O 2 , cells treated with H 2 O 2 (500 μmol/L) for 24 h; LXR-623, cells treated with LXR-623 (5 μmol/L) for 24 h; H 2 O 2 + LXR-623, cells treated with LXR-623 (5 μmol/L) for 2 h and then treated with H 2 O 2 (500 µmol/L) for 22 h; CHOL, cells were treated by cholesterol (100 μmol/L) for 24 h; H 2 O 2 + CHOL, cells were firstly treated by cholesterol (100 μmol/L) for 2 h, and then H 2 O 2 (500 µmol/L) was used to treat cells for 22 h. The scale of the images is 100 μm.

Article Snippet: All media were filled with 10% FBS, and all cell cultures were carried out in an environment at 37 °C containing 5% CO 2 , H 2 O 2 (500 μmol/L, Guangdong Hengjian Pharmaceutical Co., Ltd., Jiangmen, China), LXR-623 (5 μmol/L, HY10629, MedchemExpress, Shanghai, China), cholesterol (100 μmol/L, C8280, Solaibio, Beijing, China), tunicamycin (10 μmol/L, MB5419, Meilunbio, Dalian, China), and salinomycin (5 μmol/L, HY-15597, MedchemExpress, Shanghai, China).

Techniques: Western Blot, Staining, Control

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Cholesterol Efflux on Endothelial Dysfunction Caused by Oxidative Stress

doi: 10.3390/ijms24065939

Figure Lengend Snippet: LXR-623 can attenuate H 2 O 2 -induced endothelial cell injury by increasing cholesterol efflux, and adding cholesterol extracellularly can induce cell damage by promoting the deposition of intracellular cholesterol. ( A ) LDH, ( B ) SOD, and ( C ) NOS levels were measured with assay kits. ( D ) The activity of ROS was detected by fluorescemce probe DCFH-DA (15 μmol/L). ( E ) LDH, ( F ) SOD, ( G ) NOS levels were detected with assay kits. ( H ) Using fluorescemce probe DCFH-DA (15 μmol/L) to detect ROS activity (magnification 10×). n = 3, * p < 0.05, and ** p < 0.001 vs. the C group; # p < 0.05 and ## p < 0.001 vs. the H 2 O 2 + LXR-623 group; # p < 0.05 and ## p < 0.001 vs. the H 2 O 2 + CHOL group. The C (control) group of LXR-623 and cholesterol were treated by DMSO and chloroform, respectively; H 2 O 2 , cells treated with H 2 O 2 (500 μmol/L) for 24 h; LXR-623, cells treated with LXR-623 (5 μmol/L) for 24 h; H 2 O 2 + LXR-623, cells treated with LXR-623 (5 μmol/L) for 2 h and then treated with H 2 O 2 (500 µmol/L) for 22 h; CHOL, cells were treated by cholesterol (100 μmol/L) for 24 h; H 2 O 2 + CHOL, cells were firstly treated by cholesterol (100 μmol/L) for 2 h, and then H 2 O 2 (500 µmol/L) was used to treat cells for 22 h. The scale of the images is 100 μm.

Article Snippet: All media were filled with 10% FBS, and all cell cultures were carried out in an environment at 37 °C containing 5% CO 2 , H 2 O 2 (500 μmol/L, Guangdong Hengjian Pharmaceutical Co., Ltd., Jiangmen, China), LXR-623 (5 μmol/L, HY10629, MedchemExpress, Shanghai, China), cholesterol (100 μmol/L, C8280, Solaibio, Beijing, China), tunicamycin (10 μmol/L, MB5419, Meilunbio, Dalian, China), and salinomycin (5 μmol/L, HY-15597, MedchemExpress, Shanghai, China).

Techniques: Measured Assay, Activity Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Cholesterol Efflux on Endothelial Dysfunction Caused by Oxidative Stress

doi: 10.3390/ijms24065939

Figure Lengend Snippet: LXR-623 can attenuate the H 2 O 2 -induced adhesion activity of HUVECs by increasing cholesterol efflux, and adding cholesterol extracellularly can augment the adhesion activity of HUVECs by promoting intracellular cholesterol accumulation. ( A , C ) The expressions of ICAM-1, VCAM-1, and MCP-1 were detected by Western blot. ( B , D ) BCFECF/AM (2',7'-Di-(2-carboxyethyl)-5(6)-carboxy fluorescein) was used to label THP-1. The number of THP-1 attached to HUVEC represents the adhesion capacity of HUVEC. Green fluorescence was labeled THP-1 (magnification, 4×). n = 3, * p < 0.05 and ** p < 0.001 vs. the C group; # p < 0.05 and ## p < 0.001 vs. the H 2 O 2 + LXR-623 group; # p < 0.05 and ## p < 0.001 vs. the H 2 O 2 + CHOL group. The C (control) group of LXR-623 and cholesterol were treated by DMSO and chloroform, respectively; H 2 O 2 , cells treated with H 2 O 2 (500 μmol/L) for 24 h; LXR-623, cells treated with LXR-623 (5 μmol/L) for 24 h; H 2 O 2 + LXR-623, cells treated with LXR-623 (5 μmol/L) for 2 h and then treated with H 2 O 2 (500 µmol/L) for 22 h; CHOL, cells were treated by cholesterol (100 μmol/L) for 24 h; H 2 O 2 + CHOL, cells were firstly treated by cholesterol (100 μmol/L) for 2 h, and then H 2 O 2 (500 µmol/L) was used to treat cells for 22 h. The scale of the images is 100 μm.

Article Snippet: All media were filled with 10% FBS, and all cell cultures were carried out in an environment at 37 °C containing 5% CO 2 , H 2 O 2 (500 μmol/L, Guangdong Hengjian Pharmaceutical Co., Ltd., Jiangmen, China), LXR-623 (5 μmol/L, HY10629, MedchemExpress, Shanghai, China), cholesterol (100 μmol/L, C8280, Solaibio, Beijing, China), tunicamycin (10 μmol/L, MB5419, Meilunbio, Dalian, China), and salinomycin (5 μmol/L, HY-15597, MedchemExpress, Shanghai, China).

Techniques: Activity Assay, Western Blot, Fluorescence, Labeling, Control

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Cholesterol Efflux on Endothelial Dysfunction Caused by Oxidative Stress

doi: 10.3390/ijms24065939

Figure Lengend Snippet: LXR-623 can inhibit ER stress and the Wnt/β-catenin pathway by promoting cholesterol efflux, and adding cholesterol extracellularly can induce ER stress and activate the Wnt/β-catenin pathway by increasing the deposition of intracellular cholesterol. ( A – D ) The expressions of GRP78, CHOP, β-catenin, and p-β-catenin were detected by Western blot. n = 3, ** p < 0.001 vs. the C group; ## p < 0.001 vs. the H 2 O 2 + LXR-623 group. The C (control) group of LXR-623 and cholesterol were treated by DMSO and chloroform, respectively; H 2 O 2 , cells treated with H 2 O 2 (500 μmol/L) for 24 h; LXR-623, cells treated with LXR-623 (5 μmol/L) for 24 h; H 2 O 2 + LXR-623, cells treated with LXR-623 (5 μmol/L) for 2 h and then treated with H 2 O 2 (500 µmol/L) for 22 h; CHOL, cells were treated by cholesterol (100 μmol/L) for 24 h; H 2 O 2 + CHOL, cells were firstly treated by cholesterol (100 μmol/L) for 2 h, and then H 2 O 2 (500 µmol/L) was used to treat cells for 22 h.

Article Snippet: All media were filled with 10% FBS, and all cell cultures were carried out in an environment at 37 °C containing 5% CO 2 , H 2 O 2 (500 μmol/L, Guangdong Hengjian Pharmaceutical Co., Ltd., Jiangmen, China), LXR-623 (5 μmol/L, HY10629, MedchemExpress, Shanghai, China), cholesterol (100 μmol/L, C8280, Solaibio, Beijing, China), tunicamycin (10 μmol/L, MB5419, Meilunbio, Dalian, China), and salinomycin (5 μmol/L, HY-15597, MedchemExpress, Shanghai, China).

Techniques: Western Blot, Control