The experimental rats were anesthetized by intraperitoneal injection of 1%, 1.2 mL sodium pentobarbital (40 mg/kg of body weight) [76]. After 2 min, all anesthetized animals were immediately euthanized by cardiac blood collection just after loss of consciousness. The protocols of blood samples collection were reported previously [77, 78]. Briefly, the hair on the abdomen of animals was cut with sterile surgical scissors (Shinva Medical Instrument Co.Ltd., Shandong, China) and the skin was disinfected with 75% alcohol. Then, rats were held with left hand and kept in dorsal recumbency position at an approximately 30° angle (head lowermost). A #4 needle was inserted into the rat’s heart by the right hand at the 3rd and 4th intercostal space. About 4 mL blood sample was collected, the rat’s abdomen was quickly dissected by sterile surgical scissors and forceps (Shinva Medical Instrument Co.Ltd., Shandong, China). Then the liver and adrenal glands were collected, washed in ice-cold phosphate buffer solution (PBS) and snap-frozen immediately in liquid nitrogen. Total RNA of the liver and adrenal glands was extracted using RNA regent (HUAYUEYANG Biotechnology (Beijing) Co. Ltd) according to the manufacture’s protocol. NanoDrop 2000 (Thermo Fisher Scientific, Waltham, MA) was used to measure the concentrations and purity of RNA. The RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA) was employed to assess the integrity of RNA integrity.

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