Bc2235

For renal function assessment, lactate (BC2235, Solarbio, Beijing, China) and creatine kinase (BC1145, Solarbio, Beijing, China) were measured using assay kits according to the manufacturer’s instructions. For inflammatory cytokines, serum TNF-α (Neobioscience, Shenzhen, China), IL-1β (Solarbio, Beijing, China) and IL-6 (Neobioscience, Shenzhen, China) were determined using commercial enzyme-linked immunosorbent assay (ELISA) kits. For ELISA, data were detected by absorbance at 450 nm using a Labserv K3 Touch microplate reader (ThermoFisher, U.S.A.).
Kidney tissues was gently homogenized in homogenization solution (10 mM Tris-HCl, 1 Mm EDTA, pH 7.4). After centrifuged at 3500 g for 10 min at 4°C, the supernatant was used for the measurement of malondialdehyde (MDA) concentration (A003-1, Nanjing Jiancheng Bioengineering Institute, Jiangsu, China) and superoxide dismutase (SOD) activity (A001-1, Nanjing Jiancheng Bioengineering Institute, Jiangsu, China) using assay kits according to the manufacturer’s instructions, and IL-6 level (Neobioscience, Shenzhen, China) was determined using the ELISA kit.

Glucose concentration in culture medium and intracellular lactate concentration were determined using glucose concentration assay kit (BC2500, Solarbio) and lactic acid assay kit (BC2235, Solarbio), respectively. All these assays were performed according to the manufacturer’s protocol.

The degree of glycolysis in breast cancer cells under different conditions was detected by means of a glycolysis assay (ab197244, Abcam). Meanwhile, the degree of glucose consumption in breast cancer cells was measured using the micro method (BC2505, Solarbio). Briefly, the collected cells were centrifuged with the supernatant removed. According to the ratio of 500:1–1000:1 of the number of cells (10,000 cells): volume of distilled water (mL), the cells were broken by ultra-sonification (ice bath, power 20% or 200 W, ultrasonic 3 s, 10 s interval, repeated for 30 times) and then water-bathed at 95°C for 10 min (cover tightly to prevent water loss). After being allowed to cool down, the cells were centrifuged at 8000 × g at 25°C for 10 min, with the supernatant collected for later use. A spectrophotometer/plate reader was subsequently warmed up for >30 min, with the wavelength adjusted to 505 nm for detection purposes.
Additionally, the micro method was applied another time for detection of the degree of lactic acid production (BC2235, Solarbio), and the steps were the same as those for glucose detection. The spectrophotometer/plate reader was warmed up for more than 30 min, with the wavelength adjusted to 570 nm for detection purposes.