3100 automatic biochemistry analyzer

For biochemical analyses, the blood samples were centrifuged at 3000 rpm and 4 °C for 10 min. Serum concentrations of aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), serum total cholesterol (CHO) and serum total triglycerides (TG) were monitored by a 3100 automatic biochemistry analyzer (Hitachi Ltd., Tokyo, Japan). Insulin levels were detected by an enzyme-linked immunosorbent assay kit (Mercodia, Sweden) [29 (link)].
A liver homogenate was prepared in nine volumes of ice-cold physiological saline, and the supernatant was assayed for enzyme activities. According to Da Silva et al. [30 (link)], the supernatant of the liver homogenate was prepared for antioxidant enzyme activity under the manufacturer’s instructions of the diagnostic kits from the Nanjing Jiancheng Bioengineering Institute (China), such as glutathione peroxidase (GSH-Px) activity, catalase (CAT) activity, glutathione (GSH) and malondialdehyde (MDA) contents. The activity of cytochrome P4502E1 (CYP2E1) was measured by a CYP2E1 assay kit (Jiangsu Jingmei Biotechnology Co., Ltd., Yancheng, China) according to the instructions.

Fasting blood glucose concentrations were measured by ACCU-CHEK^® Performa (Roche, Basel, Switzerland, Germany). Plasma concentrations of cardiometabolic risk factors, including triglycerides (TG), total cholesterol (TC), HDL-cholesterol (HDL-C), and LDL-cholesterol (LDL-C) were measured using a 3100 automatic biochemistry analyzer (Hitachi, Tokyo, Japan), with enzymatic or immunoturbidimetric reagents as per the manufacturer’s protocols. Plasma concentrations of VLDL-cholesterol (VLDL-C) were calculated as VLDL-C = TG/2.2 [27 (link)], and non-HDL-cholesterol (non-HDL-C) was calculated as non-HDL-C = TC − HDL-C [28 (link)]. Plasma PCSK9 concentrations were determined with an ELISA kit (Sino Biological, Beijing, China) according to the manufacturer’s protocol.