Stress-activated gene × environment interactions may contribute to individual variability in blood pressure reductions from behavioral interventions. We investigated effects of endothelin-1 (ET-1) LYS198ASN SNP and discriminatory stress exposure upon impact of 12-week behavioral interventions upon ambulatory BP (ABP) among 162 prehypertensive African American adolescents. Following genotyping, completion of questionnaire battery, and 24-hour ABP monitoring, participants were randomized to health education control (HEC), life skills training (LST), or breathing awareness meditation (BAM). Postintervention ABP was obtained. Significant three-way interactions on ABP changes indicated that among ET-1 SNP carriers, the only group to show reductions was BAM from low chronic stress environments. Among ET-1 SNP noncarriers, under low chronic stress exposure, all approaches worked, especially BAM. Among high stress exposure noncarriers, only BAM resulted in reductions. If these preliminary findings are replicated via ancillary analyses of archival databases and then via efficacy trials, selection of behavioral prescriptions for prehypertensives will be edging closer to being guided by individual's underlying genetic and environmental factors incorporating the healthcare model of personalized preventive medicine.
Guilt and shame are usually evoked during interpersonal interactions. However, no study has compared guilt and shame processing under such circumstances. In the present study, we investigated guilt and shame in an interpersonal context using functional magnetic resonance imaging (fMRI). Behaviorally, participants reported more “guilt” when their wrong advice caused a confederate's economic loss, whereas they reported more “shame” when their wrong advice were correctly refused by the confederate. The fMRI results showed that both guilt and shame activated regions related to the integration of theory of mind and self-referential information (dorsal medial prefrontal cortex, dmPFC) and to the emotional processing (anterior insula). Guilt relative to shame activated regions linked with theory of mind (supramarginal gyrus and temporo-parietal junction) and cognitive control (orbitofrontal cortex/ventrolateral prefrontal cortex and dorsolateral prefrontal cortex). Shame relative to guilt revealed no significant results. Using multivariate pattern analysis, we demonstrated that in addition to the regions found in the univariate activation analysis, the ventral anterior cingulate cortex and dmPFC could also distinguish guilt and shame. These results do not only echo previous studies of guilt and shame using recall and imagination paradigms but also provide new insights into the psychological and neural mechanisms of guilt and shame.
• A new low toxic dual-UADLLME coupled with microwave-assisted derivatization was proposed. • 4′-Carboxy-substituted rosamine was firstly used as derivatization reagent. • Simultaneous determination of PPD and PPT in rat plasma was achieved by UHPLC-MS/MS. • This method was successfully applied to pharmacokinetics study.<br>This paper, for the first time, reported a speedy hyphenated technique of low toxic dual ultrasonic-assisted dispersive liquid-liquid microextraction (dual-UADLLME) coupled with microwave-assisted derivatization (MAD) for the simultaneous determination of 20(<b>S</b>)-protopanaxadiol (PPD) and 20(<b>S</b>)-protopanaxatriol (PPT). The developed method was based on ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) detection using multiple-reaction monitoring (MRM) mode. A mass spectrometry sensitizing reagent, 4′-carboxy-substituted rosamine (CSR) with high reaction activity and ionization efficiency was synthesized and firstly used as derivatization reagent. Parameters of dual-UADLLME, MAD and UHPLC-MS/MS conditions were all optimized in detail. Low toxic brominated solvents were used as extractant instead of traditional chlorinated solvents. Satisfactory linearity, recovery, repeatability, accuracy and precision, absence of matrix effect and extremely low limits of detection (LODs, 0.010 and 0.015 ng/mL for PPD and PPT, respectively) were achieved. The main advantages were rapid, sensitive and environmentally friendly, and exhibited high selectivity, accuracy and good matrix effect results. The proposed method was successfully applied to pharmacokinetics of PPD and PPT in rat plasma.
The article discusses hypoxic pathophysiology and high-altitude medicine as of December 2012, with a focus on the therapeutic effects of traditional Tibetan medicine Ratanasampil (RNSP) on β-amyloid (Aβ) pathologies and its potential role in Alzheimer's disease (AD) treatments. Topics include Aβ peptide production and its connection to chronic hypoxia, the use of AD transgenic mouse models, and C-terminal fragment (CTF) densitometry. Additional information is presented on the tumor necrosis factor (TNF).
OBJECTIVE: To explore the effects of traditional Tibetan medicine, Fructus Lonicerae microphyllae (FLM) on phagecytosis and cytokines production of murine macrophages.METHOD: The phagecytosis of murine macrophages was analyzed by neutral red phagecytosis assay. The activities of IL-1 and TNF-alpha were measured by biological methods. The mRNA of TNF-alpha and INF-gamma expressed by macrophages was detected by RT-PCR.
RESULT: The phagecytosis of murine macrophages was significantly enhanced by FLM at a concentration from 1 microg x mL(-1) to 100 microg x mL(-1) and the secretions of IL-1, and TNF-alpha from macrophages were markedly induced by FLM. Meanwhile, FLM also increased the expression of TNF-alpha mRNA and INF-gamma mRHA from macrophages in vitro.
CONCLUSION: FLM could promote phagecytosis and cytokines production of murine macrophages.
To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
The embryological characters of <i>Megacodon stylophorus</i> (C. B. Clark) H. Smith and <i>Veratrilla baillonii</i> Franch. are described for the first time and the systematic relationships of <i>Megacodon</i> and <i>Veratrilla</i> are discussed. Both species share the following embryological features. The anthers are tetrasporangiate. The formation of anther walls is of the Dicotyledonous Type. The tapetum is of glandular type with uninucleate cells which have a dual origin. The tapetal cells on the connective side show radial elongation or periclinal division and intrude into the anther locule to form ‘placentoids’. Cytokinesis in the microsporocyte meiosis is of the simultaneous type and the microspore tetrads are nearly always tetrahedral, rarely decussate. The ovary is bicarpellate and unilocular. The ovule is unitegmic and tenuinucellar. The formation of the embryo sac is of the Polygonum Type. Before fertilization, the two polar nuclei fuse into a secondary nucleus. Fertilization is porogamous. The development of the endosperm is of the Nuclear Type. However, the two species show variation in the following features: the number of cell layers which form the anther locule wall; construction of the wall of the mature anther; cell number in mature pollen grains; ovule number in cross sections of placentae; degree of ovule curvature; character of the hypostase and seed shape. In a comparison with the other taxa in the tribe Gentianeae using embryological features, <i>Megacodon</i> is referred to as an independent genus and should be treated as a member of the subtribe Swertiinae; <i>Veratrilla</i> is better separated from <i>Swertia s</i>.<i>l</i>. as an independent genus. <i>Veratrilla</i> is more derived than <i>Swertia s</i>.<i>s.</i> and shows a close relationship with <i>S. tetraptera</i>. © 2005 The Linnean Society of London, <i>Botanical Journal of the Linnean Society</i>, 2005, <b>147</b>, 317-331.
The embryological features of three species of <i>Swertia</i> (<i>s.l.)</i>-<i>S. erythrosticta</i>, <i>S. franchetiana</i>, and <i>S. tetraptera</i>- were characterized, and the observations were used, together with previously gathered data on other species, to evaluate a recently proposed polyphyly, based on molecular data, of <i>Swertia s.l.</i> Comparisons of species within the genus showed that they have diversified embryologically, and there are significant between-species differences. Notable features that vary between species include the number of cell layers that form the anther locule wall, the construction of the wall of the mature anther, tapetum origin, the cell number in mature pollen grains, the structure of the fused margins of the two carpels, the ovule numbers in placental cross-sections, the shape of the mature embryo sac, the degree of ovule curvature, antipodal variation and the presence of a hypostase, and seed appendages. They share characters that are widely distributed in the tribe Gentianeae, such as a dicotyledonous type of anther wall formation, a glandular tapetum with uninucleate cells, simultaneous cytokinesis following the meiosis of the microsporocytes, tetrahedral microspore tetrads, superior, bicarpellary and unilocular ovaries, unitegmic and tenuinucellar ovules, <i>Polygonum</i>-type megagametophytes, progamous fertilization, nuclear endosperm, and Solanad-type embryogeny. The presence of variation in embryological characters amongst the species of <i>Swertia s.l.</i> strongly supports the view that <i>Swertia s.l.</i> is not a monophyletic group. <i>Frasera</i> is better separated from <i>Swertia s.l.</i> as an independent genus, and is only distantly related to <i>Swertia s.s.</i> judging from the numerous differences in embryology. <i>Swertia tetraptera</i> is very closely related to <i>Halenia</i>, as they show identical embryology. © 2007 The Linnean Society of London, <i>Botanical Journal of the Linnean Society</i>, 2007, <b>155</b>, 383-400.
The characteristic fingerprint of conventional dairy Nanhanshuishi was established by X-ray diffraction (XRD), based on similarity of caculation on public peaks by MATLAB software, and the feasibility of new dairy technology of microwave method was explored between XRD and the dissolution rate in artificial simulation gastric juices. The result showed that similarity of shared peak in XRD of conventional dairy Nanhanshuishi was > 95%, This XRD characteristic fingerprint of conventional dairy Nanhanshuishi had strong specificity, could be used to provide a reference for identification and quality evaluation. This study also showed that the similarity of microware dairy products and conventional dairy products was good, and the sample of microwave 15 min was the best, and new dairy method by the microwave could replace the traditional method.
<i>Codonopsis</i>, in the family Campanulaceae, is a genus containing 42 species of dicotyledonous herbaceous perennial plants, predominantly found in Central, East and South Asia. Several <i>Codonopsis</i> species are widely used in traditional medicine and are considered to have multiple medicinal properties. Among the <i>Codonopsis</i> species, <i>Codonopsis pilosula</i> (Franch.) Nannf. and <i>C. lanceolata</i> (Sieb. et Zucc.) Benth. & Hook. f. ex Trautv. are more popular than others according to the findings, especially phytochemical and bioactive studies. Phytochemical research shows that <i>Codonopsis</i> species contain mainly polyacetylenes, phenylpropanoids, alkaloids, triterpenoids and polysaccharides, which contribute to multiple bioactivities. However, the mechanisms of their bioactivities need to be further elucidated. The less popular <i>Codonopsis</i> species remain to be studied and exploited. In addition, although a series of methods for the quality evaluation of <i>Codonopsis</i> species have been developed, a feasible and reliable approach to the efficacious and safe use of various <i>Codonopsis</i> species is still needed, with considering botanical origin, chemical constituents and bioactive effects. This review aims to provide up-to-date and comprehensive information on the phytochemistry, bioactivity and quality control of medicinal plants in the genus <i>Codonopsis</i> and to highlight current gaps in knowledge, which is useful for the wider development of the <i>Codonopsis</i> genus.
This study aims to reveal the potential relationship between 5-HT and oxidative stress in the organism. Our in vitro experiments in RIN-14B cells showed that anoxia leads the cells to the state of oxidative stress. Administration of exogenous 5-HT exacerbated this effect, whereas the inhibition of Tph1, LP533401 alleviated the oxidative stress. Several research articles reported that Cinnabar (consists of more than 96% mercury sulfide, HgS), which is widely used in both Chinese and Indian traditional medicine prescriptions, has been involved in the regulation of 5-HT. The present research revealed that HgS relieved the level of oxidative stress of RIN-14B cells. This pharmacological activity was also observed in the prescription drug Zuotai, in which HgS accounts for 54.5%, and these effects were found to be similar to LP533401, an experimental drug to treat pulmonary hypertension. Further, our in vivo experiments revealed that the administration of cinnabar or prescription drug Zuotai in zebrafish reduced the reactive oxygen species (ROS) induced by hypoxia and cured behavioral abnormalities. Taken together, in organisms with hypoxia induced oxidative stress 5-HT levels were found to be abnormally elevated, indicating that 5-HT could regulate oxidative stress, and the decrease in the 5-HT levels, behavioral abnormalities after treatment with cinnabar and Zuotai, we may conclude that the therapeutic and pharmacologic effect of cinnabar and Zuotai may be based on the regulation of 5-HT metabolism and relief of oxidative stress. Even though they aren't toxic at the present dosage in both cell lines and zebrafish, their dose dependent toxicities are yet to be evaluated.
Drawn from ancient documents of Chinese and Tibetan medicine, and including explanatory commentaries and over 700 drawings from the original texts, this book presents authentic ancient Qigong forms in an easy-to-follow format. Focusing on health practices, the selected forms also include Qigong for more esoteric purposes.
Drawing on ancient documents from China and Tibet, archaeological findings and cultural relics, this illustrated handbook presents authentic Qigong forms from the Warring States period right up the late Qing dynasty of the early twentieth century. Twenty-six sets of pictures relating to Qigong, Daoyin, diet and living habits are included, each set introduced with a brief overview of the origin, development, changes and practice modes of each method. Presented in chronological order, each chapter describes the source from which the exercises are derived, and then provides a description of the ancient form, its health and other benefits, uses, and how to do it, together with drawings of the original illustrations where these exist, or line drawings of the movements described where the original text was not illustrated. The forms are easy to learn, and easy to do, providing also a direct link to the authentic ancient forms. The selection focuses on the forms oriented to achieving resilience and general good health, but also includes many simple ancient Qigong forms for particular health problems, ranging from fatigue, indigestion and headache, to more serious conditions such as rheumatism, and even typhoid and cholera. Qigong forms directed towards the more esoteric outcomes of Qigong practice are also included. This valuable resource includes over 700 illustrations, and will be a treasure trove for all practitioners and students of Qigong, as well as anyone interested in the history of the Oriental internal arts.
OBJECTIVE: To assess the immunosuppressive effect of Artemisia vestita Wall Extract (AV-ext) on mice.METHODS: The proliferative reaction of lymphocyte and the mixed lymphocytes reaction were used to determine the effects of AV-ext on the proliferation of mouse splenocyte in vitro and in vivo; Proliferative reaction of mouse splenocyte was used for detecting the effects of AV-ext on the level of IL-2 secreted by mouse activated splenocyte in vitro. Gelatin enzymogram method and adherence analytical method were employed to disclose the effects of AV-ext on mouse activated T-lymphocytes mobility and adherence.
RESULTS: 1-100 microg/mL AV-ext exerted no obvious toxicity to mouse splenocyte, but it had obvious inhibitory effect on proliferative reaction of mouse splenocyte and mixed lymphocytes reaction induced by ConA. It also had obvious inhibitory effect on the level of IL-2 secreted by mouse activated splenocyte, on the production of MMP-9 by mouse activated T-lymphocytes, and on adherence. 150 mg/kg and 300 mg/kg of AV-ext, given to mouse per os for 7 days, could inhibit the proliferation of splenocyte and the secretion of MMP-9 by activated splenocyte of mouse.
CONCLUSION: AV-ext can inhibit the cellular immune reaction of mouse obviously.
Innovative development extends the vitality of ethnomedicines. Developing ethnomedicines is not only beneficial to the public but also to the related industry and transforms economic growth, driving local social and economic development further. Its economic benefit can be used to optimize and promote the hardware and software of the platform, as well as support the sustainable development of ethnomedicines. Apart from research and discussion on the innovative development of ethnomedicines on the basis of theory and regulations, this series of articles also summarizes cases that are conducive to the overall understanding of the necessity and feasibility of the innovative development. In terms of industrial development, large enterprises and products, such as Yunnan Baiyao, Guizhou Miao ethnomedicines, Cheezheng Tibetan Medicine, products developed from Dengzhanhua (Erigeron breviscapus), the Gold series of Yi ethnomedicines, and products developed from Sanqi (Panax notoginseng), in China are introduced and summarized, focusing on resource superiority, sustainable innovation, standard research and development, and production, as well as intellectual property protection.
Innovative development extends the vitality of ethnomedicines. Developing ethnomedicines is not only beneficial to the public but also to the related industry and transforms economic growth, driving local social and economic development further. Its economic benefit can be used to optimize and promote the hardware and software of the platform, as well as support the sustainable development of ethnomedicines. Apart from research and discussion on the innovative development of ethnomedicines on the basis of theory and regulations, this series of articles also summarizes cases that are conducive to the overall understanding of the necessity and feasibility of the innovative development. In terms of industrial development, large enterprises and products, such as Yunnan Baiyao, Guizhou Miao ethnomedicines, Cheezheng Tibetan Medicine, products developed from Dengzhanhua (Erigeron breviscapus), the Gold series of Yi ethnomedicines, and products developed from Sanqi (Panax notoginseng), in China are introduced and summarized, focusing on resource superiority, sustainable innovation, standard research and development, and production, as well as intellectual property protection.
Ethnopharmacological relevance: Saussurea laniceps Hand.-Mazz. (SL) has long been used under the herbal name Tibetan 'Snow Lotus' for the treatment of rheumatoid arthritis, stomachache and dysmenorrhea in Tibetan folk medicine. Since herbal medicine (HM) is a synergistical system with multiple components, both of the metabolism and pharmacokinetic studies of HM are interdependent. This study aimed to develop an integrated strategy based on the UPLC-DAD-QTOF-MS technique for metabolism and pharmacokinetic studies of HM.Material and methods: SL was used here as a test herb to verify the feasibility of the proposed strategy. SL was administered to rats, then, the blood plasma, urine and feces were analyzed to determine the metabolic profiles. Using our strategy, umbelliferone and scopoletin were evaluated to be the key bioactive components. Their pharmacokinetic parameters were measured and biotransformation pathways were elucidated.Results: After oral administration of SL to rats, 17 components in blood, 10 components in urine and 2 components in feces were identified and characterized using our UPLC-DAD-QTOF-MS method. Umbelliferone, scopoletin and their metabolites were found to be the major components involved in the metabolism process. Literature reports also suggest that umbelliferone and scopoletin are responsible for the therapeutic effects of SL, thus these two components were selected as the active markers for pharmacokinetic study. In the test of validity, the established method presented good linearity with R-2> 0.99. The relative standard deviation value was below 13.9% for precision, and recovery studies for accuracy were found to be within the range 91.8-112.5%.Conclusion: The present strategy offers, simultaneously, precision in quantitative analysis (metabolism study) and accuracy in quantitative analysis (pharmacokinetic study) with greater efficiency and less costs, which is therefore reliably used for integrated metabolism and pharmacokinetic studies of HM. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
OBJECTIVE: To investigate the role of Tibetan medicine-Twenty Wei Chenxiang Pill interfering with serum ET-1 level, in order to confirm that ET-1 is involved to the pathogenesis of hypoxic pulmonary hypertension.METHODS: 165 Wistar rats were randomly divided into high altitude control group,Tibetan medicine-Twenty Wei Chenxiang Pill group and plain control group. The physiological signal acquisition system was used to record pulmonary arterial pressure, and RV/(LV + S) ratio were caculated. Serum HIF-1alpha and ET-1 protein levels were determined by the method of ELISA, and ETA protein levels in lung tissue were determined by Western Blot method.
RESULTS: Compared with the high altitude group,in the rats of Tibetan medicine-Twenty Wei Chenxiang Pill group,the pulmonary arterial pressure decreased significantly from the seventh day and the seventh day (P < 0.01), the RV/(LV + S) ratio and serum HIF-1alpha levels decreased significantly from the third day (P < 0.05 or P < 0.01), the serum ET-1 levels decreased significantly from the third day (P < 0.05 or P < 0.01), and the expression of ETA protein decreased significantly from the beginning (P < 0.01 or P < 0.001).
CONCLUSION: ET-1 is one of the important factors causing pulmonary artery pressure increasing and right ventricular wall thickening, which plays a role in hypoxic pulmonary artery only involved in the early period hypoxia, but not in the later period. Tibetan medicine--twenty Wei Chenxiang Pill can prevent the pulmonary artery hypertension and the right ventricular wall thickening in rats, and its mechanism may be related to the direct inhibition of ET-1 and protein levels of ETA or the indirect downregulation of ET-1 level and ETA through inhibition of HIF-la level.
A facile I2O5-mediated direct oxidative coupling of aromatic alkenes with thiols toward vinyl sulfones has been developed under metal-free conditions. This methodology provides a convenient and efficient approach to various (E)-vinyl sulfones from readily available starting materials with excellent regioselectivity. The present oxidative coupling reaction, not only expands the scope of functionalization of alkenes with thiols, but also makes it a practical and powerful complement to traditional methods for the synthesis of (E)-vinyl sulfones.
We previously found that Ratanasampil (RNSP), a traditional Tibetan medicine, improves the cognitive function of mild-to-moderate AD patients living at high altitude, as well as learning and memory in an AD mouse model (Tg2576); however, mechanism underlying the effects of RNSP is unknown. In the present study, we investigated the effects and molecular mechanisms of RNSP on oxidative stress-induced neuronal toxicity using human neuroblastoma SH-SY5Y cells. Pretreatment with RNSP significantly ameliorated the hydrogen peroxide- (H2O2-) induced cytotoxicity of SH-SY5Y cells in a dose-dependent manner (up to 60 μg/mL). Furthermore, RNSP significantly reduced the H2O2-induced upregulation of 8-oxo-2'-deoxyguanosine (8-oxo-dG, the oxidative DNA damage marker) but significantly reversed the expression of repressor element-1 silencing transcription factor (REST) from H2O2 associated (100 μM) downregulation. Moreover, RNSP significantly attenuated the H2O2-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase 1/2 (ERK 1/2) in SH-SY5Y cells. These observations strongly suggest that RNSP may protect the oxidative stress-induced neuronal damage that occurs through the properties of various antioxidants and inhibit the activation of MAPKs. We thus provide the principle molecular mechanisms of the effects of RNSP and indicate its role in the prevention and clinical management of AD.
We previously found that Ratanasampil (RNSP), a traditional Tibetan medicine, improves the cognitive function of mild-to-moderate AD patients living at high altitude, as well as learning and memory in an AD mouse model (Tg2576); however, mechanism underlying the effects of RNSP is unknown. In the present study, we investigated the effects and molecular mechanisms of RNSP on oxidative stress-induced neuronal toxicity using human neuroblastoma SH-SY5Y cells. Pretreatment with RNSP significantly ameliorated the hydrogen peroxide- (H2O2-) induced cytotoxicity of SH-SY5Y cells in a dose-dependent manner (up to 60 μg/mL). Furthermore, RNSP significantly reduced the H2O2-induced upregulation of 8-oxo-2'-deoxyguanosine (8-oxo-dG, the oxidative DNA damage marker) but significantly reversed the expression of repressor element-1 silencing transcription factor (REST) from H2O2 associated (100 μM) downregulation. Moreover, RNSP significantly attenuated the H2O2-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase 1/2 (ERK 1/2) in SH-SY5Y cells. These observations strongly suggest that RNSP may protect the oxidative stress-induced neuronal damage that occurs through the properties of various antioxidants and inhibit the activation of MAPKs. We thus provide the principle molecular mechanisms of the effects of RNSP and indicate its role in the prevention and clinical management of AD.
We previously found that Ratanasampil (RNSP), a traditional Tibetan medicine, improves the cognitive function of mild-to-moderate AD patients living at high altitude, as well as learning and memory in an AD mouse model (Tg2576); however, mechanism underlying the effects of RNSP is unknown. In the present study, we investigated the effects and molecular mechanisms of RNSP on oxidative stress-induced neuronal toxicity using human neuroblastoma SH-SY5Y cells. Pretreatment with RNSP significantly ameliorated the hydrogen peroxide- (H2O2-) induced cytotoxicity of SH-SY5Y cells in a dose-dependent manner (up to 60 μg/mL). Furthermore, RNSP significantly reduced the H2O2-induced upregulation of 8-oxo-2′-deoxyguanosine (8-oxo-dG, the oxidative DNA damage marker) but significantly reversed the expression of repressor element-1 silencing transcription factor (REST) from H2O2 associated (100 μM) downregulation. Moreover, RNSP significantly attenuated the H2O2-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase 1/2 (ERK 1/2) in SH-SY5Y cells. These observations strongly suggest that RNSP may protect the oxidative stress-induced neuronal damage that occurs through the properties of various antioxidants and inhibit the activation of MAPKs. We thus provide the principle molecular mechanisms of the effects of RNSP and indicate its role in the prevention and clinical management of AD.
<br>Display Omitted<br>• A new coupled method of stable isotope-labeling derivatization with UA-DLLME was reported. • Simultaneous determination of multiple neurotransmitters with UHPLC-MS/MS. • Heavy labeled d3-MASC standards were used as the internal standards for quantification. • The method was sensitive, accurate and low matrix effect. • Application for neurotransmitters dynamic changes in rats brain microdialysates.<br>In this work, for the first time, a new hyphenated technique of stable isotope-labeling derivatization-ultrasound-assisted dispersive liquid-liquid microextraction has been developed for the simultaneous determination of monoamine neurotransmitters (MANTs) and their biosynthesis precursors and metabolites. The developed method was based on ultra high performance liquid chromatography tandem mass spectrometry detection using multiple-reaction monitoring mode. A pair of mass spectrometry sensitizing reagents, d0-10-methyl-acridone-2-sulfonyl chloride and d3-10-methyl-acridone-2-sulfonyl chloride, as stable isotope probes was utilized to facilely label neurotransmitters, respectively. The heavy labeled MANTs standards were prepared and used as internal standards for quantification to minimize the matrix effects in mass spectrometry analysis. Low toxic bromobenzene (extractant) and acetonitrile (dispersant) were utilized in microextraction procedure. Under the optimized conditions, good linearity was observed with the limits of detection (S/N > 3) and limits of quantification (S/N > 10) in the range of 0.002-0.010 and 0.015-0.040 nmol/L, respectively. Meanwhile, it also brought acceptable precision (4.2-8.8%, peak area RSDs %) and accuracy (recovery, 96.9-104.1%) results. This method was successfully applied to the simultaneous determination of monoamine neurotransmitters and their biosynthesis precursors and metabolites in rat brain microdialysates of Parkinson's disease and normal rats. This provided a new method for the neurotransmitters related studies in the future.
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