Rab0480

The levels of cardiac NOX1, MMP2, and TNF-α were estimated in the supernatants of the rat ventricular tissue homogenates using rat ELISA kits for TNF-α (RAB0480, Sigma-Aldrich, St. Louis, MO, USA), MMP2 (GWB-ZZD134, Genway Biotech, San Diego, CA, USA), and NOX1 (MBS2702015, MyBiosource, Inc., San Diego, CA, USA) according to the manufacturers’ instructions [68 (link)].

A quantitative chromogenic limulus amebocyte lysate assay (Aihua Medical, Shanghai, China) was performed to determine plasma endotoxin, according to the manufacturer's protocol [17 (link)]. The detection limit of endotoxin was 0.2 units, and the results are presented as EU/ml. Tumour necrosis factor- (TNF-) α levels in rat serum were detected using an ELISA kit (RAB0480, Sigma-Aldrich; Merck kGaA) and presented as ng/ml. The concentration of serum diamine oxidase (DAO), a biomarker of intestinal barrier dysfunction, was determined using ultraviolet spectrophotometry (UV-2102C, Juchne, USA).

Blood was collected by cutting the tail tips of rats at 4 weeks after surgery (n = 5 for each group). The serum β-hydroxybutyrate (BHB) levels were determined using a metabolism assay kit (MAK041, Sigma-Aldrich). Then, transcardial perfusion was conducted with a heparinized-PBS solution. SN of these rats was quickly isolated and washed using ice-cold PBS. After being dried, weighed, grinded, homogenized, and centrifuged, the obtained supernatant was analyzed to determine the levels of tumor necrosis factor-α (TNF-α, RAB0480, Sigma-Aldrich), interleukin-1β (IL-1β, AB1832P, Sigma-Aldrich), and interleukin-6 (IL-6, RAB0311, Sigma-Aldrich) in SN through ELISA kits.