Study of effects of Anmian Babu Plaster in rats with insomnia induced by p-chlorophenylalanine based on 5-hydroxytryptamine and its receptor
ZHAO Na1 XIE Chen2 LI Jinjin1 YU Xintong1 YANG Wenjia1 CHEN Yunfei1
1.Yueyang Integrative Medicine Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China;
2.Shanghai Institute of Acupuncture and Meridian, Shanghai 200030, China
Abstract：Objective To study the effects of Anmian Babu Plaster in rats with insomnia induced by p-chlorophenylalanine based on 5-hydroxytryptamine (5-HT) and its receptor. Methods A total of 48 SPF SD rats with the body weight of (200±10) g were installed with electroencephalography monitoring electrodes and divided into control group, model group, intervention group, and comfort group according to random number table method, with 12 rats in each group. Model group, intervention group, and comfort group were intraperitoneally injected with 300 mg/（kg·d） p-chlorophenylalanine for two days to make insomnia model, and control group was injected with the same amount of normal saline. After successful modeling, the control group and model group were routinely fed, shénquē in the intervention group were pasted with Anmian Babu Plaster, and shénquē in the comfort group were pasted with comfort plaster, the intervention was lasted for seven days. The sleep-wake accumulations of the four groups were observed. The levels of 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) and the expressions of 5-HT1A receptor (5-HT1AR) and 5-HT2AR in the dorsal raphe nucleus, hypothalamus, and hippocampus of the four groups were detected by enzyme-linked immunosorbent assay and immunohistochemical staining. The expression levels of 5-HT1ARmRNA and 5-H2ARmRNA were measured by RT-PCR. Results The 12 h daytime and 24 h all-day awakening volume of model group were higher than those of control group, while the rapid eye movement (REM) and non-rapid eye movement (NREM) sleep volume of model group were lower than those of control group, and the differences were statistically significant (P＜0.05). The 12 h daytime and 24 h all-day awakening volume in the intervention group were lower than those in the model group, while the REM and NREM sleep volume were higher than those in the model group, and the differences were statistically significant (P＜0.05). The levels of 5-HT, 5-HIAA, and the expression levels of 5-HT1AR mRNA in dorsal raphe nucleus, hypothalamus, and hippocampus in model group were lower than those in control group, while the expression levels of 5-HT2AR mRNA were higher than those in control group, and the differences were statistically significant (P＜0.05). The levels of 5-HT, 5-HIAA, and expression levels of 5-HT1AR mRNA in the dorsal raphe nucleus, hypothalamus, and hippocampus in the intervention group were higher than those in the model group, while the expression levels of 5-HT2AR mRNA were lower than those in the model group, and the differences were statistically significant (P＜0.05). The cell membrane staining of 5-HT1AR positive cells in the dorsal raphe nucleus, hypothalamus, and hippocampus were more deep in the control group and the intervention group, while the cell membrane staining was shallow and the number of positive cells was less in the model group. The cell membrane staining of 5-HT2AR positive cells in the dorsal raphe nucleus, hypothalamus, and hippocampus of the control group and the intervention group was shallow and the number was small; the cell membrane staining of positive cells in model group was deep and the number was large. There was no significant difference between the comfort group and the model group in each index(P＞0.05). Conclusion Anmian Babu Plaster may play a therapeutic role in rats with insomnia induced by p-chlorophenylalanine by regulating the central expression of 5-HT, 5-HT1AR, and 5-HT2AR.
赵娜1 谢晨2 李金金1 于心同1 杨文佳1 陈云飞1. 基于5-羟色胺及其受体研究安眠巴布贴对对氯苯丙氨酸致失眠大鼠的影响[J]. 中国医药导报, 2023, 20(12): 4-8.
ZHAO Na1 XIE Chen2 LI Jinjin1 YU Xintong1 YANG Wenjia1 CHEN Yunfei1. Study of effects of Anmian Babu Plaster in rats with insomnia induced by p-chlorophenylalanine based on 5-hydroxytryptamine and its receptor. 中国医药导报, 2023, 20(12): 4-8.
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