Am102 k

Triglycerides and total cholesterol were extracted from the liver tissue with chloroform-methanol (2:1, vol:vol), as previously described [21 (link)]. Briefly, chloroform-methanol was added to homogenized liver tissue, vortexed, and centrifuged; the lower phase was collected and evaporated at room temperature under a fume hood. The resulting semi-dried pellets were then dissolved in 1% Triton X-100 (VWR, USA). The hepatic and serum triglycerides (TG) and total cholesterol (TC) contents were quantified using commercial kits (Asan Pharmaceutical Co., Seoul, Korea, #AM157S-K, #AM202-K). Hepatic functional parameters such as alanine aminotransferase (ALT) and aspartate transaminase (AST) activities were measured using commercial kits (Asan Pharmaceutical Co., Seoul, Korea, #AM102-K, #AM103-K). Serum concentrations of TNF-α and IL-6 were determined using Quantikine ELISA kits (R&D Systems, Minneapolis, MN, USA, #MTA00B, #M6000B).

Mouse blood was harvested from one-side eyeball while the mice were in deep anesthesia and then stored at −80 °C. Serum samples were extracted by centrifuge. Biochemical parameters, aspartate aminotransferase (AST, AM102-K, Asan Pharmaceutical, Seoul, Korea), alanine aminotransferase (ALT, AM103-K), blood urea nitrogen (BUN, BUN Serum Detection Kit, ARBOR ASSAYS, Ann Arbor, MI, USA), and creatinine (Creatinine Serum Detection Kit, ARBOR ASSAYS, MI, USA) were detected according to the manufacturers’ manuals.

Serum aspartate aminotransferase (AST/GOT, glutamate-oxaloacetate transaminase) and alanine aminotransferase (ALT/GPT, glutamate pyruvate transaminase) were measured using GOT reagents (AM103-K; Asan Pharmaceutical, Seoul, Korea) and GPT reagents (AM102-K; Asan Pharmaceutical) according to the manufacturer’s instructions.
Serum levels of 17β-estradiol, TNF-α, and IL-6 were measured using a mouse ELISA kit of 17β-estradiol (ES180S-100; Calbiotech, El Cajon, CA, USA), IL-6 (M6000B; R&D systems), TNF-α (MTA00B; R&D system) according to the manufacturer’s instructions.
The levels of hepatic LXA₄ were measured using mouse LXA₄ ELISA kit (CSB-E13268m, Cusabio, Wuhan, China) following to manufacturer’s protocols.