Abstract: Objective To observe the regulating effects of drug-containing serum of Li-pi compound preparation on LP,NPY and CCK-8 proteins and genes in rat hypothalamic neuron cells and to explore the mechanism of its treatment for anorexia.Methods Fifty rats were randomly divided into control group, low-, medium- and high-dose Li-pi compound preparation group and zinc sulfate group, with 10 in each group. Control group was given equal volume of drinking water by gavage. Zinc sulfate group was given 20 mg/kg zinc sulfate oral solution. Low-, medium- and high-dose Li-pi compound groups were given 2.4,4.8 and 9.8 g/kg of the compound, respectively. Each group was given gavage once a day for 7 days in a row, and the femoral artery blood was collected on the 7th day after gavage to prepare the drug-containing serum. Rat hypothalamic neuron cells were cultured in vitro and identified by MAP2 immunofluorescence. The hypothalamic neurons were randomly divided into control group, low-, medium- and high-dose Li-pi compound preparation group, and zinc sulfate group by CCK-8, and effect on the activity of hypothalamic neuron cells was assessed at 0 h, 2 h,4 h,6 h,8 h,12 h,24 h,36 h and 48 h. After 24 hours of the corresponding serum, the activity of hypothalamic neurons in each drug-containing serum group began to increase and became different. ELISA was used to detect LP, NPY and CCK-8 content in supernatant of hypothalamic neuron cells; Western Blot and real-time fluorescence quantitative PCR were used to detect the relative expression of LP, NPY and CCK-8 protein and genes in hypothalamic neuronal cells.Results The cells were identified as hypothalamic neuronal cells. Compared with the control group, the LP,NPY and CCK-8 content in supernatant of hypothalamic neuron cells was decreased in the supernatant of hypothalamic neuron cells of low-，medium- and high-dose Li-pi compound preparation, and the differences were statistically significant(P<0.05);the relative expression of LP,NPY and CCK-8 protein in hypothalamic neuron cells in Li-pi compound preparation of medium and high doses was decreased, and the differences were statistically significant(P<0.05); the relative expression of LP gene in hypothalamic neuron cells in high-dose Li-pi compound preparation group was decreased, and the difference was statistically significant(P<0.05); the relative expression of NPY and CCK-8 genes in hypothalamic neuron cells in low-, medium- and high-dose Li-pi compound preparation groups was decreased, and the differences were statistically significant(P<0.05).Conclusion Li-pi compound preparation may improve anorexia-related symptoms by comprehensively regulating the expression of LP, NPY and CCK-8 proteins and genes in hypothalamic neurons, but its effect is negatively correlated with dose.

Key words:

Li-pi compound preparation, Hypothalamus, Neuron cells, Primary culture, LP, NPY, CCK-8