Acetonitrile and formic acid

The following reagents and instruments were used for this study: 7600 automatic biochemical analyzer from Japan's Hitachi company, blood coagulation analyzer from German BE company, Wasters UPLC-Q Exactive HF MS (Thermo Fisher Scientific, Rockford, IL, USA), centrifuge (Microfuge 22, Beckman Coulter company), vortex mixer (Vortex Genius 3, German IKA group), Acetonitrile and formic acid (from the Fisher company in the United States), ammonium acetate, and analytically pure (from the Sigma-Aldrich company in the United States), and ultrapure water was prepared by the MILLI-Q ultra purified water preparation system (US Millipore Corporation).

A 2,2-diphenyl-1-picrylhydrazyl (DPPH•), 2,20-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), Folin–Ciocalteu reagent, 2,20-azobis (2-methylpropionamide) dihydrochloride (AAPH), Trolox, acetonitrile and formic acid (both MS grade), berberine (95% purity), and ferulic acid were purchased from Sigma–Aldrich (Stenheim, Germany). Ascorbic acid was purchased from Stanlab (Lublin, Poland), and analytical-grade methanol and aluminium chloride hexahydrate were purchased from POCH (Gliwice, Poland). Ultrapure water was prepared with a Milli-Q purification system (Millipore, Burlington, MA, USA).
The SuperScript Vilo cDNA Synthesis kit was used according to the manufacturer’s recommendations. cDNA was stored at −20 °C until subsequent analysis. PCR reactions were performed using SG onTaq qPCR Master Mix (Eurx, Gdańsk, Poland) on the QuantStudio 5 Real-Time PCR System (Applied Biosystems, Waltham, MA, USA).