Astragalin

Mulberry leaves were collected from Longling county (Baoshan, China) and identified to be the leaves of Mours alba by Prof. Baolian Guo, the Institute of Medicinal Plant Development, Peking Union Medical College. The insulin detection kit was purchased from Wuhan Miting Biotechnology Co., Ltd. (Wuhan, China). Assay kits for triglyceride (TG), total cholesterol (T-CHO), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were purchased from Jiancheng Bioengineering Institute (Nanjing, China). Streptozotocin was purchased from Sigma Co., (Saint Louis, MO, USA). Standard mouse diet was purchased from Mouse One Mouse Two Biological Co., Ltd. The antibodies for AMPK, SREBP1, ACC, CPT1A, FAS, and horseradish peroxidase-conjugated goat-anti-rabbit IgG were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Other chemicals were supplied by Damao Co., Ltd. (Tianjin, China). Standards of rutin, hyperoside, and astragalin were purchased from Chengdu Must Bio-Technology Co., Ltd. (Chengdu, China).

Reference substances of hyperin, neochlorogenic acid, chlorogenic acid, p-coumaric acid, astragalin, isoquercitrin, and formononetin (IS) with purity over 98.0% were achieved from Chengdu Must Biotechnology Co., Ltd. (Chengdu, China). The acetonitrile and formic acid were obtained from Dikma (Foothill Ranch, CA, USA). Ultra-pure water was provided by Milli-Q system (Millipore, Boston, MA, USA).

Astragalin (purity ≥98%) was purchased from Chengdu Must Bio-Technology Co., Ltd. (Sichuan, China) and dissolved in dimethyl sulfoxide (Sigma) before use. The dimethyl sulfoxide concentration in the working solutions was <0.1%, which had no effect on the present study. Terminal deoxynucleotidyl nick-end labeling (TUNEL) assay was conducted using in situ cell death detection kit (POD, Roche, Germany). All other reagents were of standard biochemical quality and were obtained from commercial suppliers.