Icarisid II rescues cognitive dysfunction via activation of Wnt/ß-catenin signaling pathway promoting hippocampal neurogenesis in APP/PS1 transgenic mice.

Restoring the compromised neurogenesis has been served as a potential strategy to rescue cognitive dysfunction of Alzheimer's disease (AD). In this study, we explored whether icarisid II (ICS II), a natural product possessing powerful neuroprotection, could recover the neurogenesis dysfunction of APP/PS1 mice, and investigated its underlying mechanisms. Our results showed that oral administration of ICS II could alleviate cognitive injuries of APP/PS1 mice, promote hippocampal neurogenesis, as well as stimulate Wnt/ß-catenin signal pathway confirmed by upregulated Wnt-3a, phosphorylated glycogen synthase kinase-3ß (p-GSK-3ß), and ß-catenin. ICS II also depressed mitochondrial fission evidenced by upregulated Mitofusin 1 (Mfn 1) and Mitofusin 2 (Mfn 2), and downregulated mitochondrial fission 1 protein (Fis 1), mitochondrial fission factor (Mff), and phosphorylated dynamin-related protein 1 (p-Drp 1). However, these effects of ICS II were blunted by XAV-939, an inhibitor of Wnt/ß-catenin signaling pathway. In summary, our findings revealed that ICS II could improve neurogenesis and inhibit mitochondrial fission via activation of the Wnt/ß-catenin signaling pathway, which contributed to cognitive function restoration of APP/PS1 mice. This study discovered a novel mechanism involving neurogenesis regulation underlying the therapeutic effects of ICS II against AD.

Icarisid II promotes proliferation and neuronal differentiation of neural stem cells via activating Wnt/ß-catenin signaling pathway.

Adult neurogenesis plays a vital role in maintaining cognitive functions in mammals and human beings. Mobilization of hippocampal neurogenesis has been regarded as a promising therapeutic approach to restore injured neurons in neurodegenerative diseases including Alzheimer's disease (AD). Icarisid II (ICS II), an active ingredient derived from Epimedii Folium, has been reported to exhibit multiple neuroprotective effects. In the present study, we investigated the effects of ICS II on the proliferation and differentiation of neural stem cells (NSCs) and amyloid precusor protein (APP)-overexpressing NSCs (APP-NSCs) in vitro. Our results demonstrated that ICS II dose-dependently suppressed apoptosis and elevated viability of APP-NSCs. ICS II (1 µM) potently promoted proliferation and neuronal differentiation of NSCs and APP-NSCs. ICS II (1 µM) significantly upregulated Wnt-3a expression, increased the phosphorylation of glycogen synthase kinase-3ß and enhanced the nuclear transfer of ß-catenin. Moreover, ICS II also promoted astrocytes to secrete Wnt-3a, which positively modulates Wnt/ß-catenin signaling pathway. These findings demonstrate that ICS II promotes NSCs proliferation and neuronal differentiation partly by activating the Wnt/ß-catenin signaling pathway.

[Comparison of 12 constituents of Astragali Radix directionally processed with organic acid by UPLC-MS].

To establish an UPLC-MS method for the simultaneous content determination of 4 saponins and 8 flavonoids, in order to analyze the effect of Astragali Radix directionally processed with organic acid on the content of glycosides and aglycones. The separation was carried out on ACQUITY UPLC HSS T_3(2.1 mm×100 mm, 1.8 µm), the mobile phase was eluted with the mixture of 0.1 mol·L~(-1) formic acid water solution and 0.1 mol·L~(-1) formic acid acetonitril in a gradient mode. The detection wavelength was 260 nm, the flow rate was 0.5 mL·min~(-1), the column temperature was 30 ℃, and the injection volume was 2 µL. Mass spectrometry analysis was performed with an electrospray ionization(ESI) source in a positive ion mode. The 12 constituents showed good linear relations within their own ranges(R~2≥0.999 2),with good average recoveries. The results showed no significant change in saponins but both qualitative and quantitative changes in flavonoids after directional processing of Astragali Radix with organic acid. The established method can provide methodological reference for analyzing the effect of Astragali Radix directionally processed with organic acid on glycosides and aglycones.

[Verification of processing theory of "reducing ketone and dryness, and increasing ester and effect" for bran-fried atractylodes].

We previously proposed the processing theory of "reducing ketone and dryness, and increasing ester and effect" for bran-fried atractylodes, and made a preliminary study. To further verify the science and rationality of the theory, we determined the changes in the content of atractylenolide I, II, III and atractylon in atractylodes after and before being fried with bran, in order to compare the effect of raw and bran-fried atractylodes on the water intake and urination in rats in this study. The effect of raw and bran-fried atractylodes on the content of four gastrointestinal hormones and two neurotransmitters in serum was observed in an attempt to verify the science and rationality the processing theory of "reducing ketone and dryness, and increasing ester and effect" for bran-fried atractylodes.

Simultaneous Determination of Five Chromones of Radix Saposhnikoviae Extract in Rat Plasma by UPLC-MS/MS: Application to a Comparative Pharmacokinetic Study in Normal and Febrile Rats.

A rapid and sensitive quantitative analytical method was established for the simultaneous determination of five chromones (prim-O-glucosylcimifugin, cimifugin, 4'-O-ß-D-glucosyl-5-O-methylvisamminol, 5-O-methylvisammiol, and sec-o-glucosylhamaudol) in the plasma of RS-treated rats for the first time using ultra performance liquid chromatography- (UPLC-) tandem mass spectrometry. The Waters Acquity UPLC BEH C18 (50 mm × 2.1 mm, 1.7 µm) was used as the chromatographic column, 0.1% formic acid water and 0.1% formic acid acetonitrile comprised the mobile phases, and all samples were determined under positive ion mode. The results showed that all analytes had good linearity (r > 0.9902), between-day and within-day precisions less than 15%, accuracy between -5.50% and 5.53%, and extraction recovery between 88.26% and 97.65%. Both the matrix effect and stability met the requirements. This method was successfully applied for the comparative pharmacokinetics of five active components of RS in normal and febrile rats. The results showed that the pharmacokinetic behavior of RS extract significantly differed between the two types of rats.

Metabolomics analysis and rapid identification of changes in chemical ingredients in crude and processed Astragali Radix by UPLC-QTOF-MS combined with novel informatics UNIFI platform.

Astragali Radix, the root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge., is widely used as a tonic decoction pieces in the clinic of traditional Chinese medicine (TCM). Astragali Radix has various processed products with varying pharmacological actions. There is no modern scientific evidence to explain the differences in pharmacological activities and related mechanisms. In the present study, we explore the changes in chemical components in Astragali Radix after processing, by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with novel informatics UNIFI platform and multivariate statistical analysis. Our results showed that the crude and various processed products could be clearly separated in PCA scores plot and 15 significant markers could be used to distinguish crude and various processed products by OPLS-DA in UNIFI platform. In conclusion, the present study provided a basis of chemical components for revealing connotation of different processing techniques on Astragali Radix.

Metabolomic study of raw and processed Atractylodes macrocephala Koidz by LC-MS.

Bai-Zhu, the dried rhizome of Atractylodes macrocephala Koidz (AMK), is widely used as a tonic herbal in eastern Asia. It is commonly used as prepared slices in clinic by stir-frying with wheat bran (processed AMK). In the theories of traditional Chinese medicine (TCM), Bai-Zhu possesses significantly different therapeutic effects before and after processing. However, the molecular mechanics of this processing is still unknown. In this paper, the strategy of metabolomics was employed to investigate the changes of chemical constituents in Atractylodes macrocephala Koidz after processing. Meanwhile, the cell activity test variation of processed and unprocessed medicine was used to interpret the processing mechanism of AMK. Using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF/MS) with the method of multivariate statistic analyses including principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA), atractylenolide I, atractylenolide II, atractylenolide III, atractylenolide VI, 7-hydroxycoumarin, 8-ß-methoxy atractylenolide I and Selina-4 (14), 7 (11)-dien-8-one were rapidly explored as the potential chemical markers of raw and processed AMK, respectively. Furthermore, it could be speculated that the processing mechanism of AMK was to increase the content of atractylenolide III which could strengthen the effect on gastrointestinal function.

Determination of orcinol glucoside by LC-MS in Curculigo orchioides and its application to a pharmacokinetic study

ABSTRACT This study was designed to explore the pharmacokinetic regularity of the plasma concentration, tissue distribution and excretion of orcinol glucoside from aqueous extracts of raw and processed Curculigo orchioides Gaertn., Hypoxidaceae. The experiment first used an ultrahigh-performance liquid chromatography-tandem mass spectrometry approach with multiple reaction monitoring and a positive mode to separate orcinol glucoside from naringin to obtain the plasma concentration curves, bar graph of tissue distribution and excretion curves. These results might be beneficial for reasonable clinical application of C. orchioides and for further development of its wine and salt-processing mechanism.