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Introduction: To explore the mechanism of action of appling Radix Ginseng and Semen Ziziphi Spinosae Drug pair (R-S) in the treatment of insomnia by investigating the effect of R-S on GLU/GABA-GLN metabolic cycle and intestinal microflora of rats with insomnia. Methods: Rats were intraperitoneally injected with 4-chloro-DL-phenylalanine (PCPA) to make sleep deprivation (SD) models. The rats were divided into 6 groups, with 8 rats in each group. The general status of the rats was observed and the pentobarbital sodium sleep synergy experiment was performed. The contents of GABA, GLU, GLN, GAD65, and GS in hippocampus of rats were determined by ELISA. The expressions of GABAARα1mRNA, mGluR5mRNA, NR1mRNA and GluR1mRNA in rats' hippocampal tissue were determined by Realtime PCR. 16SrRNA gene sequencing was used to analyze the intestinal microflora of insomnia rats. Results: In PCPA-induced insomnia rats, the state of insomnia was relieved, the sleep rate was improved, the duration of sleep latency was shortened and the sleep duration was prolonged in each dose group of R-S (p < 0.05, p < 0.01) compared with the model group. The contents of GABA, GLN, GAD65 and GS were increased (p < 0.05, p < 0.01) while GLU content was decreased (p < 0.01) in both medium and high dose groups, especially in the high dose group. The expression of GABAARα1mRNA was increased (p < 0.01), and the expressions of mGluR5mRNA, NR1mRNA and GluR1mRNA were decreased (p < 0.01) in hippocampal tissue of rats in R-S groups, especially in the high dose group. At the same time, the various dose groups of R-S could improve the species diversity, microflora abundance of insomnia rats and regulate the KEGG metabolic pathway related to sleep. Discussion: R-S can improve the sleep of PCPA-induced insomnia rats by regulating GLU/GABA-GLN metabolic cycle and intestinal microflora, which provides experimental basis for appling R-S in the treatment of insomnia.
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BACKGROUND: The rhizome of Atractylodes lancea (AL) is usually used for the treatment of various diseases such as spleen deficiency syndrome (SDS). Both bran-processed and crude AL is included in Chinese Pharmacopoeia. The different efficacies of bran-processed and crude AL on SDS are largely unknown, and the mechanisms of AL effects have not been fully elucidated. OBJECTIVE: The objective of the study was to compare the effects of bran-processed and crude AL and then assess the mechanisms of treating SDS. MATERIALS AND METHODS: The model of SDS in rats was established using excessive exertion, combined with an irregular diet and intragastric administration of the extract of Sennae Folium, and different doses of bran-processed and crude AL were gavaged. The serum was analyzed by an enzyme-linked immunosorbent assay (ELISA), and small intestinal tissues were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The injury of SDS was alleviated by the treatment of bran-processed and crude AL. Compared to model group, the indexes of trypsin (TRY), amylase (AMS), vasoactive intestinal peptide (VIP), somatostatin (SS), gastrin (GAS), substance P (SP), Na+-K+-ATPase, and succinic dehydrogenase in serum of each administration group were increased by ELISA, and the mRNA expressions of VIP, SS, GAS, and SP in small intestinal tissues were increased by RT-PCR. Furthermore, in a dose-dependent manner, the bran-processed and crude AL increased the levels of TRY, AMS, VIP, and GAS and the mRNA expression levels of VIP. Compared with the crude AL, the bran-processed AL was more effective in treating SDS. CONCLUSION: Through the mechanisms of treating SDS by AL, both bran-processed and crude AL has alleviated the symptoms of SDS. SUMMARY: Both bran-processed and crude Atractylodes lancea (AL) alleviated symptoms of spleen deficiency syndrome (SDS)Comparing with crude AL, bran. processed AL was more effective in treating SDSThe efficacy of AL could be partly attributed to digestive enzyme activity, gastrointestinal hormone levels, membrane protein activity, and changes in mitochondrial activity. Abbreviations used: AL: Atractylodes lancea; TRY: Trypsin; AMS: Amylase; VIP: Vasoactive intestinal peptide; SS: Somatostatin; GAS: Gastrin; SP: Substance P; ELISA: The enzyme-linked immunosorbent assay; mRNA: Messenger ribonucleic acid; SDH: Succinic dehydrogenase; RT-PCR: Reverse transcription-polymerase chain reaction; TCM: Traditional Chinese medicine; SDS: Spleen deficiency syndrome.
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To study the influence of astragaloside on mRNA expression of PI3K/Akt/mTOR signal transduction in anemia model mice induced by chemotherapy, 48 male BALB/c mice which were 6-7 week old were picked as the research objects and randomly divided into four groups, blank group, model group, astragaloside group and astragaloside IV group. Each group was 12 mice. Chemotherapy anemia model was established by cyclophosphamide. The mice were drawn blood from eyeball after 14 days treatment. The QPCR was used to test the mRNA concentrations of Akt, PI3K, BCL-xl, bad, FoxO, mTOR, PTEN in mouse spleen. In comparison of blank group, astragaloside group and astragaloside IV group,the erythrocyte counting and values of Hb in model group were significantly lower (P<0.05). The volumes mRNA of Akt,PI3K,BCL-xl,bad,mTOR were lower in blank group, compared with other groups (P<0.05 or P<0.01). The similar trend in astragaloside IV group except PI3K, comparing with blank group (P<0.05 or P<0.01). The contents of these five genes were no significant differentiations between astragaloside group and blank group. The statistics were obvious between astragaloside group and astragaloside IV group (P<0.05 or P<0.01). The concentrations of FoxO, PTEN were higher in model group,compared with blank group and astragaloside group (P<0.05 or P<0.01), but no difference with astragaloside IV group. Comparing with blank group, the volumes of these two genes were increased in astragaloside IV group (P<0.05), FoxO was higher in astragaloside group (P<0.05), but PTEN was not significant. There was no the same as astragaloside group and Astragaloside IV group. Therefore, astragaloside could increase the contents of Akt, PI3K, BCL-xl, bad, mTOR (P<0.01), decrease the concentrations of FoxO, PTEN (P<0.05). The changes in cyclophosphamide-induced anemia were highly significant by astragaloside. It could be related to the mRNA expression of PI3K/Akt/mTOR Signal Transduction.
