Peg400

PEG
400, acetonitrile,
and methanol were procured from Sigma-Aldrich (Mumbai, India), CDH,
and Spectrochem (India), respectively. Sodium acetate and acetic acid
were procured from Sigma-Aldrich, Mumbai, India. The acetate buffer
was freshly prepared for dissolution and analytical studies. Solvents
were of analytical reagent grade (AR). Millipore water was used as
an aqueous medium for varied PEG 400 to water ratio in the mixture.

Cardiac dysfunction was induced by an intraperitoneal injection of LPS (10 mg/kg; cat. no. L2880; Sigma-Aldrich; Merck KGaA) for 24 h, as previously described (14 (link)). ATX (40 mg/kg; cat. no. SML0982; Sigma-Aldrich; Merck KGaA) dissolved in polyethylene glycol 400-N, N-dimethylacetamide (PEG400; 1:1 v/v; cat. no. 91863; Sigma-Aldrich; Merck KGaA) was administered to the mice in the LPS + ATX group 30 min after LPS administration for 24 h. The mice in the control and LPS groups were administered an equivalent volume of the vehicle (PEG400; 1:1 v/v; cat. no. 91863; Sigma-Aldrich; Merck KGaA). The health, behavior and death of mice were monitored every 6 h following LPS administration. In the LPS and LPS + ATX groups, 8 and 3 mice died, respectively, of severe infection following LPS treatment. All the surviving animals were anesthetized with pentobarbital solution (50 mg/kg) via an intraperitoneal injection 24 h after LPS treatment. Following anesthesia, 0.5 ml blood was collected from the mice by eyeball enucleation; which prompted two mice to die from severe anemia. To isolate myocardial tissue, mice were sacrificed by cervical dislocation. Following a 2 min observation, no respiration and corneal reflex in the mice confirmed death. The heart tissue was isolated post-mortem and stored in 4% paraformaldehyde or at −80°C.

C57BL/6 male mice (six-week-old, Charles River, Wilmington, MA) were randomly divided into control and treatment groups (n = 8 for each group). The mice in the control group were treated with a modified AIN-93G diet (Table S1) containing 0.5% v/w polyethylene glycol 400 (PEG 400, EMD Millipore, Billerica, MA), and the mice in the treatment group were treated with the diet containing TCC (99%, Sigma-Aldrich, St. Louis, MO) dissolved in PEG 400 during the whole experiment. After 3 weeks, the mice were treated with 2% DSS (36–50 kDa, MP Biomedicals, Solon, OH) in drinking water to induce acute colitis. After 9 days, the mice were sacrificed, and the blood and colon tissues were collected for analysis.