Citric acid sodium citrate buffer

The T2DM model was induced as follows: Experimental animals were fed basal chow for 1 week. After 1 week, the diabetic group was fed high-fat and high-sugar chow, and the control group was fed basal chow. At week 5, all animals were fasted for 12 h (without water), and STZ (1%) was dissolved in 0.1 mmol/L citric acid-sodium citrate buffer (pH 4.2; Solarbio, Beijing, China), fully dissolved and stored on ice away from light. A single rapid intraperitoneal injection of 30 mg/kg streptozotocin (MCE, USA) was administered to the diabetic group. An equal volume of 0.1 mM citric acid-sodium citrate buffer was intraperitoneally administered to the control group. Seven days later, blood was drawn from the tail vein to check for glucose levels of the rats in the diabetic group. Rats with blood glucose ≥ 16.7 mmol/L were fed a high-fat, high-sugar diet for another 8 weeks, while rats that did not meet the blood glucose standard were discarded. Blood glucose was retested after 8 weeks, and modeling was successful if blood glucose was still ≥ 16.7 mmol/L [16 (link)].

BCL-2 (ab196495) and GAPDH (#2881S) antibodies were obtained from Abcam (Cambridge, United Kingdom), whereas Caspase-3 (#9662S) antibody was purchased from Cell Signaling Technology (Danvers, MA, United States). TGF-β (A18692) antibody was purchased from Abclonal (Wuhan, China), and Bax (ET1603-34), MMP9 (ER1706-40), MyHC (ET1702-88), and P2X7R (ER1901-99) antibodies were obtained from Huabio (Hangzhou, China). The above antibodies were diluted at 1:1,000. In addition, Collagen 1 (sc293182) and ANP (sc-515701), which were diluted at 1:200, were obtained from Santa Cruz Biotechnology (Dallas, TX, United States). Goat anti-rabbit secondary antibodies (A0208) and goat anti-mouse secondary antibodies (A0216) which were used in the Western blot and One-Step TUNEL Apoptosis Assay Kit, respectively, were purchased from Beyotime (Shanghai, China). Hematoxylin-eosin (H&E) Staining Kit and Masson’s Trichrome Stain Kit were obtained from Solarbio (Beijing, China). STZ was purchased from Sigma (CA, United States) and citric acid-sodium citrate buffer was purchased from Solarbio (Beijing, China).