ABSTRACT
Objective:To study the protective effect and possible mechanism of psoralen corylifolia on non-alcoholic steatohepatitis (NASH) induced by high-fat diet in mice.Methods:The newly weaned female mice in the offspring of C57BL/6J mice fed with normal diet were selected as the control group (gavage of distilled water); the newly weaned female mice in the offspring of C57BL/6J mice fed with high-fat diet were randomly divided into model group (gavage distilled water), low-dose group[psoralen corylifolia 1.125 mg/(g·d)], high-dose group [psoralen corylifolia 2.25 mg/(g·d)] and vitamin E group [vitamin E 0.01 mg/(g·d)]. Six mice in each group were fed continuously for 8 weeks. Automatic biochemical analyzer was used to detect serum alanine aminotransferase (ALT), aspartate transaminase (AST), triglyceride (TG), total cholesterol (TC) level in mice; The liver tissue pathological changes were observed by hematoxylin-eosin (HE) and Sirius-red (SR) staining; The level of reactive oxygen species (ROS) in liver tissue was detected by dihydroethidium (DHE) fluorescence probe; the activity of NADPH oxidase was detected by kit; The protein expressions of nuclear factor-κB (NF-κB), phosphatidylinositol 3 kinase (PI3K p85), protein kinase B (Akt), P47 phox and protein kinase C-α (PKC-α) were detected by Western blot.Western blot. Results:The levels of serum ALT, AST, TG, TC and homeostasis model assessment of insulin resistance (HOMA-IR) index in the model group were higher than those in the control group (all P<0.01). After treatment, the levels of serum ALT, AST, TG, TC and HOMA-IR in low-dose group, high- dose group and vitamin E group were lower than those in model group (all P<0.05). HE and SR staining showed that hepatocytes in the model group were swollen, and there were lipid droplets of different sizes, vacuoles and obvious fibrosis. After treatment, hepatocyte steatosis and fibrosis decreased and the contents of ROS and NADPH oxidase in liver decreased(all P<0.05); Western blot showed that the p-p65/p65, p-Akt/Akt, p-PKC-α/PKC-α, PI3K, p85 and P47 phox protein expression in the model group were higher than those in the control group (all P<0.01). After treatment, the protein expression levels of p-p65/p65, p-Akt/Akt, p-PKC-α/PKC-α, PI3K, p85 and P47 phox decreased (all P<0.01). Among the above indexes, the protective effect of high-dose group on liver NASH was better than those of vitamin E group and low-dose group (all P<0.05). Conclusions:Psoralen corylifolia can improve the liver function of NASH model mice, which may be related to the inhibition of oxidative stress, inflammatory reaction and liver fibrosis, which provides a new idea for the prevention and treatment of children with NASH.
ABSTRACT
Objective: To establish an accurate and selective UPLC-MS/MS) method for the determination of afatinib in rat plas-ma. Methods: Protein precipitating by acetonitrile was used to prepare the samples. A CORTECS BEH C18column ( 50 mm × 2. 1 mm, 1. 6 μm) was used to separate the analytes at 40℃. The mobile phase consisted of acetonitrile and water (0. 1% formic acid) with the flow rate of 0. 4 ml·min-1. The analytes were quantified by multiple reaction monitoring ( MRM) mode with positive electrospray ionization, while the target fragment ions were m/z 486. 19→112. 1 for afatinib and m/z 557. 3→112. 15 for neratinib (IS). Results: The calibration curve obtained good linearity for afatinib within the range of 1–200 ng·ml-1(r=0. 998 1), and the LLOQ in rat plasma was 1. 0 ng/ml. The intra-and inter-day precisions were both≤9. 51% . The recovery of afatinib from plasma was above 77. 1% . After intragastric administration and intravenous administration of afatinib in rats, the t1/2was 7. 19 h and 2. 69 h, Cmax was 97. 78 ng·ml-1and 123. 37 ng·ml-1,and AUC(0-∞)was 1 505. 4 ng·ml-1·h and 405. 55 ng·ml-1·h, respectively. Con-clusion: The validated method can be applied in the pharmacokinetic study of afatinib at the intragastric and intravenous dosage of 10 and 2 mg·kg-1, respectively.