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Regulation of Hypnotic Active Components of Cnidii Fructus on Melatonin Synthesis Rate-limiting Enzyme AANAT in Rats with PCPA-induced Insomnia / 中国实验方剂学杂志
Article in Chinese | WPRIM | ID: wpr-906329
Responsible library: WPRO
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 reactionReal-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.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article