Effects of LW-AFC, a new formula derived from Liuwei Dihuang decoction, on intestinal microbiome in senescence-accelerated mouse prone 8 strain, a mouse model of Alzheimer disease / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effects of LW-AFC, a new formula derived from Liuwei Dihuang decoction, on gut microbiota and the behavior of learning and memory of SAMP8 mice, a mouse model of Alzheimer Disease (AD), and identify the specific intestinal microbiota correlating with cognitive ability. METHODS Morris-water maze test, novel object recognition test and shuttle-box test were conducted to observe the ability of learning and memory. 16S rRNA amplicon sequencing (Illumina, San Diego, CA, USA) was employed to investigate gut microbiota. RESULTS The treatment of LW- AFC improved cognitive impairments of SAMP8 mice, including spatial learning and memory ability, active avoidance response, and object recognition memory capability. Our data indicated that there were significantly 8 increased and 12 decreased operational taxonomic units (OTUs) in the gut microbiota of SAMP8 mice compared with senescence accelerated mouse resistant 1 (SAMR1) strains, the control of SAMP8 mice. The treatment of LW- AFC altered 22 (16 increased and 6 decreased) OTUs in SAMP8 mice and among them, 15 OTUs could be reversed by LW-AFC treatment resulting in a microbial composition similar to that of SAMR1 mice. We further showed that there were 7 (3 negative and 4 positive correlation) OTUs significantly correlated with all the three types of cognitive abilities, at the order level, including Bacteroidales, Clostridiales, Desulfovibrionales, CW040, and two unclassified orders. LW-AFC had influences on bacterial taxa correlated with the abilities of learning and memory in SAMP8 mice and restored them to SAMR1 mice. CONCLUSION The effects of LW-AFC on improving cognitive impairments of SAMP8 mice might be via modulating intestinal microbiome and LW-AFC could be used as a potential anti-AD agent.

[HPLC fingerprint of glycyrrhizea radix et rhizoma praeparata cum melle].

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.

Danggui-Shaoyao-San Improves Learning and Memory in Female SAMP8 via Modulation of Estradiol.

Previous studies showed that Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription, could alleviate cognitive dysfunction of Alzheimer's disease (AD) patients. However, the mechanisms remain unclear; we have now examined the effect of DSS on SAMP8 and elucidated the possible mechanism. Animals were treated with DSS for 2 months, and step-down test and Morris water maze (MWM) test were used to evaluated cognitive abilities. The estradiol (E2), NO, and glycine in blood plasma or in hippocampus were detected to explore the possible mechanisms. The latency of SAMP8 in step-down test was shorter than that of age-matched SAMR1, and DSS increased the latency especially in female animals. In MWM test, we got similar results; SAMP8 spent more time to find the platform, and DSS decreased the time before finding the platform, with little effect on swim velocity, during the training sessions. During test session, DSS increased the time spent in target quadrant especially in female SAMP8. In female SAMP8, plasma E2, NO, and glycine were elevated in plasma or hippocampus tissue. In conclusion, DSS could ameliorate deterioration of cognition in SAMP8, especially in female animals. Increasing E2, NO, and glycine might contribute to the cognitive improvement effect of DSS in female SAMP8.

[Chemical constituents of glucoside fraction from Liuwei Dihuang Gantang].

OBJECTIVE: To study chemical constituents of glucoside fraction from Liuwei Dihuang Gantang and clarify its substance foundation of active constituents. METHOD: Glucoside fraction was prepared by macroporous resin chromatography. Constituents were separated by silica gel and reverse phase silica gel column chromatography, and their structures were identified by MS and NMR. RESULT: Eleven compounds were separated and identified as 7-dehydrologanin (1), 7alpha-O-methylmorroniside (2), 7beta-O-methylmorroniside (3), 7alpha-O-ethylmorroniside (4), 7beta-O-ethylmorroniside (5), morroniside (6), sweroside (7), loganin (8), paeoniflorin (9), paeonolide (10) and loganic acid (11). CONCLUSION: All of those compounds were separated from Liuwei Dihuang Gantang for the first time.

[HPLC fingerprint of Liuwei Dihuang condensed pills].

OBJECTIVE: To establish HPLC fingerprints of Liuwei Dihuang condensed pills. METHOD: Dikma Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) was adopted, with acetonitrile (containing 0.05% phosphoric) -water (containing 0.05% phosphoric) as the mobile phase. The column temperature was set at 40 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was 276 nm (0-10 min), 236 nm (10-40 min) and 276 nm (40-60 min). The sample size was 20 microL. Chromatographic peaks were identified by Q-TOF-MS-IDA-MS/MS method. RESULT: Good precision, stability and repeatability were proved. Q-TOF-MS-IDA-MS/ MS method was adopted for qualitative determination of eighteen chromatographic peaks. Ten batches of Liuwei Dihuang condensed pills were determined with the method, and their similarities were above 0. 96. CONCLUSION: The study lays a foundation for the overall quality evaluation of Liuwei Dihuang condensed pills.

JD-30, an active fraction extracted from Danggui-Shaoyao-San, decreases ß-amyloid content and deposition, improves LTP reduction and prevents spatial cognition impairment in SAMP8 mice.

JD-30 is an active fraction extracted from Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription. We previously showed that JD-30 could alleviate cognitive dysfunction of the mice induced by intracerebroventricular injection of ß-amyloid (Aß). However, data remain scarce on the effect of JD-30 on an Alzheimer's disease (AD) model and the underlying mechanisms are unknown. Further detailed studies on the effects of JD-30 on spatial cognition of senescence-accelerated mouse prone 8 (SAMP8), a suitable rodent model for cognitive impairment of aged subjects were investigated to elucidate the possible mechanisms. Long-term treatment with JD-30 significantly decreased the prolonged latency of SAMP8 in the Morris water-maze test. It also ameliorated the reduction of long-term potentiation (LTP) and reduced the damage of neurons in the hippocampus of SAMP8. Finally, JD-30 decreased the content and deposition of Aß in the brain of SAMP8. The results show that JD-30 improves deterioration of spatial learning and memory in the SAMP8 mouse model, and by decreasing the content and deposition of Aß, neuronal activity and synaptic plasticity improve, suggesting one of the mechanisms involved.

Danggui-Shaoyao-San and its active fraction JD-30 improve Abeta-induced spatial recognition deficits in mice.

AIMS OF THE STUDY: Previous studies showed that Danggui-Shaoyao-San (DSS), a traditional Chinese medicinal prescription, could alleviate cognitive dysfunction of Alzheimer's disease (AD) patients. However, the mechanism and substance basis remain unknown. JD-30 is a fraction extracted from DSS, whose activity we previously was evaluated. beta-Amyloid (Abeta) is believed to be a critical etiological factor of AD. We have now examined the effect of DSS and JD-30 on AD model mice induced by Abeta, and elucidated the possible mechanism. MATERIALS AND METHODS: Mice were intracerebroventricular injected with the aggregated Abeta(25-35) to mimic AD. Groups of mice were treated with DSS or JD-30 by intragastric infusion over 2 weeks, and their spatial learning and memory capacities were measured by the Morris water maze procedure. The mechanisms were investigated by extracellular microelectrode recordings, and also electron microscopy. RESULTS: Our results show that Abeta(25-35) induced impairment of spatial learning and memory in mice, as well as inhibition of long-term potentiation (LTP) in the hippocampus. The impairments were ameliorated by DSS or JD-30 administration. Additionally, JD-30 not only prevented the aggregation of Abeta(25-35), but disrupted aggregated Abeta(25-35) fibrils. CONCLUSION: These results suggest that JD-30 is one of the chief active fractions extracted from DSS by its ability to ameliorate deterioration of cognition, and by blocking and disrupting the aggregation of Abeta so that synaptic plasticity was improved, which may be one of the mechanisms involved.

[A new monoterpene glycoside from active fraction (DSS-A-N-30) of Danggui Shaoyao San].

OBJECTIVE: To study the chemical constituents of an active fraction (DSS-A-N-30) from Danggui Shaoyao San. METHOD: DSS-A-N30 was prepared by macroporous resin chromatography, the compound was isolated by column chromatography on silica gel and RPC-18, the structure was elucidated by spectroscopic methods. RESULT: A new monoterpene glycoside was isolated and identified from DSS-A-N-30. CONCLUSION: The new monoterpene glycoside was identified as 4"-hydroxyl-albiflorin.

[Determination of eleutheroside B in antifatigue fraction of Acanthopanax senticosus by HPLC].

OBJECTIVE: To develop an HPLC method for the determination of eleutheroside B in As1 (the extract fraction from Acanthopanax senticosus, which has a good effect of antifatigue). METHOD: The antifatigue effect of Asl was evaluated by mice burden swimming testing. One compound was isolated by silica gel, Sephadex LH-20 column chromatography and preparative high performance liquid chromatography. The structure was identified by physicochemical properties and spectral evidences. Eleutheroside B in Asl was determined by HPLC. Chromatographic conditions included Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase consisting of a mixture of acetonitrile-water-ethanoic acid (10: 90: 0.01). The UV detection wavelength was set at 344 nm. RESULT: As1 showed an excellent antifatigue activity; One compound was isolated from As1 and its structure was identified as Eleutheroside B; The calibration cure was linear in the range of 0.104-20.8 microg (r = 0.9999), the average recovery was 97.68%, RSD 1.4% (n=6). CONCLUSION: This HPLC method is simple,accurate and reliable.

[Studies on bioassay-guided anti-inflammatory fraction in bark of Albizia julibrissin combined determination with LC-MS-MS].

OBJECTIVE: To search the anti-inflammatory fraction of Albizia julibrissin. METHOD: Inflammatory model of Kunming mice ear edema induced by croton oil and determination combined with the LC-MS-MS-guided fractionation and isolation were used. RESULT: The n-butanol fraction (AJ-B) obtained from the ethanolic extract of the Cortex albiziae was the major active fraction. The lignan glycosides fraction (AJ-B-1), which was further isolated from AJ-B, showed significant anti-inflammatory activity and exhibited dose-dependent relationship in the dose of 5 to 20 mg x kg(-1). CONCLUSION: The method of bioassay-guided fractionation and isolation combined with the LC-MS-MS determination may be of benefit to the logical studies on the bioactive fractions or constituents of traditional Chinese materia medica.

[Studies on chemical constituents from stellaria media. I].

OBJECTIVE: To study the chemical constituents of Stellaria media. METHOD: The compounds were isolated with various column chromatography and semi-preparative HPLC. The structures were determined by modem spectroscopic methods (IR, UV, 1H-NMR, 13C-NMR, HMBC, ESI-MS). RESULT: Five compounds were isolated and identified as apigenin 6-C-beta-D-galactopyranosyl-8-C-alpha-L-arabinopyranoside (1), apigenin 6-C-alpha-L-arabinopyranosyl-8-C-beta-D-galactopyranoside (2), apigenin 6-C-beta-D-galactopyranosyl-8-C-beta-L-arabinopyranoside (3), apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-galactopyranoside (4), apigenin 6, 8-di-C-alpha-L-arabinopyranoside (5). CONCLUSION: Compounds 1-5 were obtained from Stellaria genus for the first time.

[Advances on the study of Danggui Shaoyao powder].

As a traditional Chinese medicine, Danggui Shaoyao powder has been studied for more than one thousand years. This article reviews the recent advances in chemical constituents, pharmacological activities and clinical application.

Determination of rizatriptan in human plasma by liquid chromatographic-eletrospray tandem mass spectrometry: application to a pharmacokinetic study.

A sensitive liquid chromatographic-tandem mass spectrometry(LC-MS/MS) method was developed for the determination of rizatriptan in human plasma. The analytes were extracted from plasma samples by liquid-liquid extraction, separated on a Zorbax XDB C8 column (150 x 4.6 mm i.d.) and detected by tandem mass spectrometry with an electrospray ionization interface. Zomitriptan was used as the internal standard. The method had a lower limit of quantitation of 50 pg/mL for rizatriptan, which showed more sensitivity and speed of analysis compared with reported methods. The within- and between-day precision was measured to be below 11.71% and accuracy between -5.87 and 0.86% for all quality control samples. This quantitation method was successfully applied to the evaluation of the pharmacokinetic profiles of rizatriptan after single oral administration of 5, 10 and 15 mg rizatriptan tablets to 10 healthy volunteers (five males and five females).

[Advances in study on bark of Albizzia julibrissin].

Cortex Albizziae, the stem bark of the leguminous plant Albizzia julibrissin, is specified in Chinese pharmacopoeia as a traditional Chinese medicine used to relieve melancholia and uneasiness of body and mind, invigorate the circulation of blood and subside a swelling. This article reviews the recent advances in chemical constituents and pharmacological activities of Cortex Albizziae.

High-performance liquid chromatography combined with mass spectrum analysis for identifying the antifatigue components in Acanthopanax senticosus Harms.

The roots of Acanthopanax senticosus (Rupr. et Maxim.) Harm were obtained for the acquisition of the crude extractives to isolate the 19 components by means of preliminary extraction and isolation. High-performance liquid chromatography followed by primary and secondary mass spectrum analysis (HPLC/MS/MS) was employed to identify the antifatigue components from the extracts of the plant with reference to the literature. Good isolation results were achieved and several known and unknown chemical compounds were identified, for instance, in one sample (No. 11) 5 known components were detected along with 6 unknown compounds with relative molecular mass of 302, 318, 346, 354, 390, and 406, respectively.

[Determination of glycosides in traditional Chinese medicine liu-wei di-huang by RP-HPLC].

OBJECTIVE: To establish a RP-HPLC method for determination of glycosides in Traditional Chinese Medicine Liu-wei Di-huang. METHOD: The samples were analyzed on an ODS column at 30 degrees C, with mobile phase of methanol/water (33:67) at flow rate 1.0 mL.min and detection at wavelength of 236 nm. RESULT: Three major components reached base-line separation and were identified to be mononiside, loganin, paeoniflorin. Respectively for the three components, linear correlations were found between peak areas and concentrations in the ranges of 7.4-60, 7.7-62 mg.L-1 and 8.5-68 mg.L-1, and the recoveries were 98.8%, 98.3%, 99.6%. CONCLUSION: The established method is proved to be suitable for simultaneous quantification of three major glycosidic components in Liuwei Dihuang decoction and can be used for evaluation of the quality of Liuwei Dihuang preparations.