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Article in Chinese | WPRIM | ID: wpr-906329

ABSTRACT

Objective:To observe the effects of Cnidii Fructus hypnotic active components (CHC) on the behaviors of rats with p-chlorophenylalanine (PCPA)-induced insomnia and melatonin (MT) synthesis rate-limiting enzyme arylalkylamine <italic>N</italic>-acetyltransferase (AANAT), and explore the protective mechanism of CHC on the pineal gland. Method:Male SD rats of SPF grade were randomly divided into a blank control group, a model group, a MT group, and high-, medium-, and low-dose CHC groups with 10 rats in each group. Except for the blank control group, other groups received 4.5% PCPA suspension at 10 mL·kg<sup>-1</sup>, intragastric administration, for two consecutive days. After PCPA model of insomnia was established, normal and model groups were gavaged at the same volume of 2% Tween-80, MT control group (10 mg·kg<sup>-1</sup>), CHC was high, medium and low (60, 30, 15 mg·kg<sup>-1</sup>), 10 mL·kg<sup>-1</sup>, once a day, for consecutive 7 days. Four days after administration, open field, elevated cross maze, and pentobarbital sodium-induced sleep tests were conducted, respectively. Serum MT was detected by enzyme-linked immunosorbent assay. The mRNA expression level of AANAT was determined by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR). The expression of AANAT protein in the pineal gland was detected by Western blot. Result:Compared with the results in the blank control group, the total distance of open field activity and standing times and duration in the central area were increased (<italic>P</italic><0.05, <italic>P</italic><0.01), the proportions of open arm entry (OE%) and open arm time (OT%) were decreased (<italic>P</italic><0.05), and the sleep latency was prolonged (<italic>P</italic><0.01) in the model group. Compared with the model group, no significant difference was observed in the low-dose CHC group, while other groups exhibited reduced total distance of activity (<italic>P</italic><0.05, <italic>P</italic><0.01), elevated OE% (<italic>P</italic><0.05), shortened sleep latency, and prolonged sleep time (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with the serum MT in the blank control group, that in the model group was decreased (<italic>P</italic><0.01). Compared with the model group, no significant difference was observed in the low-dose CHC group, while other groups displayed increased serum MT (<italic>P</italic><0.05). The mRNA and protein expression of AANAT was decreased in the model group as compared with that in the blank control group (<italic>P</italic><0.01). Compared with the model group, the MT group and the high-dose CHC group showed up-regulated expression (<italic>P</italic><0.05). Conclusion:CHC improved the behavioral indexes of PCPA-induced insomnia, increased the synthesis and secretion of MT in pineal cells, and elevated the serum MT level, which was related to the up-regulation of the mRNA and protein expression of AANAT in the pineal gland.

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