C57bl 6j mice

Specific-pathogen-free (SPF) male Wistar rats (weighting 200 ± 10 g) were purchased from Qingdao Peitefude rat breeding Co., Ltd. (Qingdao, China, license number: SCXK(Lu)20130004). Eight-week-old male and female C57BL/6J mice (SPF, 22–25 g) were obtained from Beijing Huafukang Bioscience Co., Ltd. (Beijing, China, license number: SCXK(Jing)20140004). Animals were housed in cages under standardized conditions: a 12-h light/dark cycle, 23 ± 2°C room temperature, and 55 ± 5% relative humidity. Animals had free access to food and water. The Ethics Committee of Qingdao University approved all the experimental procedures, and animals were treated according to the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The ethical committee approval number of this project is 2017HC11LQ076.

All animal experiments were performed with the approval of the Institutional Animal Care and Use Committee of Peking University First Hospital (Approval Number: J201534). Male KK-Ay mice (N = 50) and C57BL/6J mice (N = 10) were purchased from Beijing HFK Bioscience Co., Ltd. [License No. SCXK (Jing) 2012-0001]. The mice were aged 8–9 weeks old and weighed 30 ± 2 g. Blood glucose was measured via the glucose oxidase method. All KK-Ay mice with PPG ≥11.1 mmol/L or FG ≥7.0 mmol/L and MA tested in the urine were used as DN samples, and all others were excluded. The mice recruited were randomly divided into six groups as follows: (1) control (untreated C57BL/6J mice); (2) model (untreated KK-Ay mice); (3) metformin (KK-Ay mice treated with 250 mg/kg metformin); (4) low JTD (KK-Ay mice treated with 2 g/kg ethanol extract of JTD); (5) medium JTD (KK-Ay mice treated with 4 g/kg ethanol extract of JTD); and (6) high JTD (KK-Ay mice treated with 8 g/kg JTD ethanol extract). All drugs were administered orally once per day for 12 weeks.

JUN mice were provided by the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences, and housed in a specific pathogen‐free (SPF) and standard barrier environment with free access to food and water. The JUN mice bred up to 47 generations and 6 generations in our breeding environment. C57BL/6J mice purchased from Beijing Huafukang Bioscience company were used as controls. Both females and males had significant phenotypes, so the sex of the mice used in the study was random. All procedures in this study involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College (GR20004).

Male KK-Ay mice (n = 69, 3-month-old) and age-matched male C57BL/6J mice (n = 10) were purchased from HFK Bioscience Co. Ltd. (Beijing, China). KK-Ay mice were fed a high-fat diet. Age-matched male C57BL/6J mice fed with the standard chow diet were employed as non-diabetic control by oral gavage distilled water. Male KK-Ay mice were randomly divided into four groups: untreated group (n = 19) and treated groups. Treated groups received PHPB (50 mg/kg/day, n = 17 and 150 mg/kg/day, n = 17, Yunnan Haobang Pharmaceutical Co. Ltd., Yunnan, China) [13 (link)] or Rosiglitazone (5 mg/kg/day, n = 16, Chongqing Taiji Pharmaceutical Co., Ltd., Chongqing, China) [15 (link),42 (link)] by oral gavage. All treatment continued for two months, beginning at 3-month-old, and body weight was monitored throughout the experiment.
During the experimental period, animals were allowed free access to food and water ad libitum in a temperature-controlled environment of 23 ± 2 °C and maintained at a 12-h light/dark cycle per day. All experiments were approved and performed in accordance with the guidelines for the care and use of laboratory animals and were approved by the Animal Care Committee of the Peking Union Medical College and the Chinese Academy of Medical Sciences (Beijing, China) (Approval number 00000634).

Forty male C57BL/6 J mice (aged 8 weeks old; weighing 20–22 g) were purchased from HFK Bioscience (Beijing, China). The mice were kept in a room at a relative humidity of 55% ± 10%, a temperature of 23°C ± 2°C, and a light/dark cycle of 12 h. The mice were allowed free access to drinking water and standard laboratory food. Animals and all study protocols were performed strictly in accordance with Chinese standards and guidelines for the use of laboratory animals (GB14925-2001 and MOST 2006 a), and were carried out with the approval of the Experimental Animal Welfare Ethics Committee of the North Sichuan Medical College (Approval No. 202205).