Mechanism-based anti-anxiety effects of polysaccharides extracted from shudihuang (radix rehmanniae preparata) by two-dimensional electrophoresis analysis in rat hippocampus proteins.

OBJECTIVE: To investigate mechanism-based anti-anxiety effects of Shudihuang (Radix Rehmanniae Preparata) polysaccharides (RRPPs) through two-dimensional electrophoresis (2-DE) analysis with mass spectrometry (MS) of hippocampus proteins in rats treated with monosodium L-glutamate (MSG). METHODS: MSG (4 g/kg) or normal saline (NS) was injected subcutaneously into infant male rats on days 2, 4, 6, 8, 10 after birth. MSG-treated rats at 8 weeks old were given NS, diazepam, or RRPPs daily for seven consecutive days by intragastric administration, while NS-treated rats given the same volume of NS. Elevated plus maze (EPM) and light/dark transition (LDT) tests were used to observe anti-anxiety effects of RRPPs at 1 h after the last administration. After EPM and LDT tests, hippocampus tissues were excised on ice rapidly from the brains of rats. Thereafter, 2-DE and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) were used for detecting differential proteins in hippocampus of rats so as to explore the potential mechanisms. RESULTS: RRPPs (200, 400 mg/kg) could significantly inhibit MSG-induced decrease of time and entries percentages in open zones in EPM test and numbers of light-dark transition in LDT test. Further analysis of 2-DE and MALDI-TOF/MS indicated that beta-synuclein, protein DJ-1, peroxiredoxin-2, peroxiredoxin-6, dimethylarginine dimethylaminohydrolase 1 (DDAH-1) and iron-sulfur proteins were all found to be down-regulated significantly in MSG-treated rats, while such down-regulation was significantly inhibited after treatment with RRPPs. CONCLUSION: RRPPs showed anti-anxiety effects and potential mechanisms might be related to inhibiting MSG-induced down-regulation of beta-synuclein, DJ-1, peroxiredoxin-2, peroxiredoxin-6, DDAH-1 and iron-sulfur proteins in hippocampus of rats.

[Effects of Semen Ziziphi Spinosae on the anxiety behavior of the yin deficiency mice].

OBJECTIVE: To observe the effects of Semen Ziziphi Spinosae on the anxiety behavior of the yin deficiency mice, to filter the active material of Semen Ziziphi Spinosae and explore its anti-anxiety mechanism. METHODS: The yin deficiency mice model with intragastrical 320 mg/kg thyroid tablet for nine days exception normal control group. To observe yin deficiency model mice anxiety action by means of the elevated plus-maze and light-dark transitions of mice. After the experiment, peeled off cerebra at once. The left saved in liquid nitrogen for assaying GABA and Glu with TLCS, and right saved in formalin for the protein expression of GABAaR1 and NMDAR1 with immunohistochemical method. RESULTS: (1) Compared with yin deficiency model group, Tabellae Diazepami group and the middle alcohol extracts group increased instinctively in the percentage of time and degree entering open-arm in eleveated plus maze, and the passing times in light-dark transitions increased instinctively (P<0.05). (2) Compared with yin deficiency model group, the contents of GABA increased evidently (P<0.05) and Glu reduced evidently (P<0.05) in Tabellae Diazepami group and the middle alcohol extracts group. (3) Compared with yin deficiency model group,the expression of GABAAR1 increased evidently (P<0.05) and NMDAR1 reduced evidently (P<0.05) in the diszepam group and the middle alcohol extracts group. CONCLUSIONS: The alcohol extracts of Semen Ziziphi Spinosae have the anti-anxiety effects instinctively. Its mechanism may be related to increasing the GABA and expression of GABAAR1 and reducing the Glu and expression of NMDAR1.