Raloxifene

Recombinant human cytochrome P450 isoenzymes (rhCYP) were purchased from Cypex (Dundee, UK). The human liver microsomes (50-Donor, LOT: IHG) were obtained from Bioreclamation IVT (MD, USA). d-Glucose-6-phosphate (G-6-P, LOT: WXBC5003V), glucose-6-phosphate dehydrogenase (LOT: SLBJ9913V), NADP sodium salt (LOT: BCCD0008), sulfaphenazole (LOT: 083H0050) were obtained from Sigma–Aldrich (MO, USA). Raloxifene (LOT: J0206A), sulconazole (LOT: N1211AS), fluvastatin (LOT: J0701A), quinidine (LOT: M0110A), omeprazole (LOT: D0929As), clomethiazole (LOT: D1225A), 8-methoxypsoralen (LOT: M0113A), tetraethylenepentamine (TEPA, LOT: S0801A), ticlopidine (LOT: A0606AS) were purchased from Meilun Biotechnology (Dalian, China). Prothrombin time assay kit purchased from Nianjing Jiancheng Bioengineering Institute (Nanjing, China). The purity of DDAO and HXMB was >98%. All other reagents were of the highest grade commercially available.

This study was divided into four groups according to different combinations of drugs: 10^-7 M 17-beta estradiol (Abbott Healthcare Products B.V.) +10^-9M raloxifene (Dalian Mei Lun Biotechnology Co., Ltd.) as the TSEC group, 10^-7M 17-beta estradiol + 3.3×10^-8 M medroxyprogesterone acetate (Beijing Yimin Pharmaceutical Co., Ltd.) as the E+P group, 10^-7M 17-beta estradiol alone as the E group, and a control group (13 (link),29 (link),30 (link)). After different treatment, the proliferation and apoptosis of primary cultured uterine leiomyoma cells were analyzed by methylthiazolyldiphenyl-tetrazolium bromide (MTT) and western blot.

Cisplatin (molecular formula: Cl2H6N2Pt, molecular weight: 300.05) was purchased from Jiangsu Hansoh Pharmaceutical Group Co. Ltd., and raloxifene (molecular formula: C28H27NO4S.HCl, molecular weight: 510.05) from Dalian Meilun Biotechnology Co. Ltd. RIPA (radio immunoprecipitation assay) buffer, phenylmethanesulfonyl fluoride (PMSF), BCA protein quantification kit, and the superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH) detection kits, JC-1 mitochondrial membrane potential assay kit and 5-Ethynyl-2ʹ-deoxyuridine (EdU) Cell Proliferation Kit with Alexa Fluor 594 were purchased from Beyotime Biotechnology. Hematoxylin and eosin (H&E) staining solutions, cell counting kit 8 (CCK-8) were purchased from KeyGEN Biotechnology Co. Ltd. Periodic acid-Schiff (PAS) staining solutions and corn oil were purchased from Solaibao Technology Co. Ltd., ethanol and xylene from Aladdin reagents, and the creatinine and urea nitrogen detection kits from Kehua Biological Engineering Co. Ltd. Primary antibody: anti-Bcl-2, anti-Bax, anti-β-actin were purchased from Wanleibio Company (Shenyang, China). HRP-labeled secondary antibody was purchased from Proteintech Company (Wuhan, China).