Creatinine

Ergosterol (ERG, purity 98%) was purchased from National Institutes for Food and Drug Control (Beijing, China). Allopurinol (ALL) tablets were acquired from Simcare Drug Store (Nanjing, China). Streptozotocin (STZ) was provided by Sigma (St Louis, MO, USA). Insulin enzyme-linked immunosorbent assay (ELISA) kits were supplied by Nanjing KeyGEN Biotech. Co., Ltd. (Nanjing, China). Glucose, triglycerides (TG), total cholesterol (TC), uric acid, and creatinine commercial kits were purchased from Jiancheng Bioengineering Institute (Nanjing, China). Primary antibodies against PI3K (p85 subunit), p-PI3K (p85 subunit), Akt, p-Akt, NF-κBp65, p-NF-κBp65, IкBα, and p-IкBα were produced by Cell Signaling Technology Inc. (Beverly, MA, USA).
All other chemicals and reagents used for study were of analytical grade and were purchased from approved organizations.

Animals were fasted overnight, and blood was collected in heparin-containing tubes by means of heart puncture under anaesthesia. Blood samples were centrifuged at 3000 RPM for 10 min at 4 °C and stored at − 80 °C until analysis. The plasma concentrations of creatinine (#C011–2-1, Nanjing Jiancheng, China), aminotransferase (#C009–2 and #C010–2, Nanjing Jiancheng, China), total cholesterol (#A111–1-1, Nanjing Jiancheng, China), high-density lipoprotein cholesterol (#A112–1-1, Nanjing Jiancheng, China), low-density lipoprotein cholesterol (#A113–1-1, Nanjing Jiancheng, China), and triglycerides (#A110–1-1, Nanjing Jiancheng, China) were measured enzymatically using an automated chemical analyser. The levels of plasma cytokines, including monocyte chemoattractant protein-1 (MCP-1, #SW2544505), soluble tumour necrosis factor receptor 2 (STNFR2, #SW2544507), tumour necrosis factor-α (TNF-α, #YX-201407 M), interleukin-6 (IL-6, #YX-091206 M), interleukin-1β (IL-1β, #YX-091203 M), LPS (#YX-121619 M), D-lactose (#SW2544501), and diamine oxidase (DAO) (#SW2544502), were analysed with ELISAs according to the manufacturer’s instructions (Sino Best Biological Technology Co., Ltd., China).

Mice were administered with 0.2 mL saline containing 12 mg fluorescein isothiocyanate-4 kDa dextran, 8 mg rhodamine B isothiocyanate-70 kDa dextran, and 20 mg creatinine (Sigma) 3 h prior to the end of 24 h and 28 days PS micro- and nanoplastics exposure. Serum was harvested after 24 h of PS micro- and nanoplastics exposure. creatinine was measured using a kit from Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). Recovery of creatinine and fluorescent probes was measured in a Synergy HT plate reader (BioTek, Winooski, VT, USA) using freshly prepared standards. Fluorescein and rhodamine B fluorescence were measured using excitation wavelengths of 495 nm and 555 nm, and emission wavelengths of 525 nm and 585 nm, respectively.

Potassium oxonate, HypoXanthine, Xanthine, and XO were purchased from Shanghai yuan ye Bio-Technology Co., Ltd. (Shanghai, China). Assay kits for the determination of urea nitrogen (BUN), creatinine (CRE), Aspartate aminotransferase (AST), and Alanine aminotransferase (ALT) were obtained from Nanjing Jiancheng Bioengineering Inc., (Nanjing, China) Biotechnology. 0.9% NaCl injection was obtained from Anhui Shuanghe Pharmaceutical Co., Ltd. (Anhui, China). The uric acid Kit was purchased from Biosino Biotechnology and Science inc (Beijing, China). HPLC-grade methanol and acetonitrile were purchased from Merck (Beijing, China). Ultrapure water was prepared by A.S. Watson TM Limited (Beijing, China).

Misai Kuching, North Cordyceps, Longan Arillus, Folium Ginseng, and Corni Fructuswere purchased from the Yunnan Chinese Herbal Medicine Department. These herbs were authenticated by one of our authors, Professor Ziren Su, Guangzhou University of Chinese Medicine (GZUCM). The voucher specimens (Yu 15-6-25) were kept at School of Chinese Materia Medica, GZUCM, for reference. D-gal and vitamin E (VE) were purchased from Sigma-Aldrich (St. Louis, USA). Commercial biochemical assay kits for the measurements of superoxide dismutases (SOD), catalase (CAT), glutathione peroxidase (GPx), malondialdehyde (MDA), alanine aminotransferase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), and creatinine (CRE) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Tween 80 was purchased from Sinopharm (Shanghai, China). All reagents used were of either analytical or chromatographic grade.

GTPs (91.21% catechins and 71.72% EGCG) were purchased from Corona Science & Technology Development Co. Ltd. (Fu Zhou, China). Rabbit polyclonal antibodies (anti-MnSOD, anti-HMGCS2, anti-Nampt, and anti-catalase) were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, USA). Anti-FOXO3a and anti-SIRT3 antibodies were from Cell Signaling Technology Inc. (Danvers, USA). Anti-SOD2/MnSOD (acetyl K122) and anti-4-HNE antibodies were from Abcam Inc. (Cambridge, UK). Total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and blood glucose were detected using kits from BioSino Bio-Technology & Science Inc. (Beijing, China). ELISA kits for the quantitative measurement of serum cystatin C and insulin were from Biovendor Inc. (Heidelberg, Germany) and Mercodia AB (Uppsala, Sweden). Creatinine and N-acetyl-β-D-glucosaminidase (NAG) assay kits were purchased from the Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). DMEM/high glucose was purchased from Gibco Inc. (New York, USA). Lipofectamine 3000 transfection reagent was from Invitrogen Inc. (California, USA).