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To build up an identification method on cardiac glycosides in Taxillus chinensis and its Nerium indicum host, and evaluate the influence on medicine quality from host to T. chinensis, ultra-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass-mass spectrometry(UPLC-Q-TOF-MS/MS)was applied. The samples of T. chinensis(harvested from N. indicum)and its N. indicum host were collected in field. The samples of T. chinensis(harvested from Morus alba)and its M. alba host was taken as control substance. All samples were extracted by ultrasonic extraction in 70% ethanol. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 C_(18)(2.1 mm×100 mm,1.8 μm)column at 40 ℃. Gradient elution was applied, and the mobile phase was consisted of 0.1% formic acid water and acetonitrile. The 0.5 μL of sample solution was injected and the flow rate of the mobile phase was kept at 0.6 mL·min~(-1) in each run. It was done to identify cardiac glycosides and explore the chemical composition correlation in T. chinensis and its N. indicum host by analyzing positive and negative ion mode mass spectrometry data, elemental composition, cardiac glycoside reference substance and searching related literatures. A total of 29 cardiac glycosides were identified, 28 of it belonged to N. indicum host, 5 belonged to T. chinensis(harvested from N. indicum host), none of cardiac glycoside was identified in T. chinensis(harvested from M. alba host). The result could provide a reference in evaluating the influence in T. chinensis medicine quality from host. It was rapid, accurate, and comprehensive to identify cardiac glycosides in T. chinensis and its N. indicum host by UPLC-Q-TOF-MS/MS.
Subject(s)
Cardiac Glycosides , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Loranthaceae , Chemistry , Nerium , Chemistry , Phytochemicals , Tandem Mass SpectrometryABSTRACT
Objective: To establish a method for identifying cardiac glycosides in Scurrula parasitica and its Nerium indicum host by UPLC-Q-TOF-MS/MS. With safflower parasitoids with sweet-scented osmanthus trees as the host and their host osmanthus tree samples used for control, the chemical constituents of the cardiac glycosides were identified by comparison between the cardiac glycoside reference substances and literatures, so as to analyze the correlation between the safflower parasitoid and its host oleander glycoside components,and evaluate the host' s impact on the quality of Taxilli Herba. Method: Samples of S. parasitica (parasitic on N. indicum and Osmanthus fragrans),N. indicum and O. fragrans were collected. Samples of S. parasitica and its O. fragrans host were taken for control. All of the samples were extracted through ultrasonic extraction with 70%ethanol. ACQUITY UPLC HSS T3 C18(2.1 mm×100 mm,1.8 μm) column was adopted with mobile phase A comprising 0.1%formic acid water and mobile phase B comprising acetonitrile for gradient elution. The sample size was 0.5 μL. The flow rate was 0.6 mL·min-1. The column temperature was maintained at 40℃. MassLynx V4.1 software was used to analyze the data. Identification and correlation of chemical constitute of cardiac glycosides in S. parasitica and its N. indicum host were performed through analysis on cardiac glycosides reference substances,relevant literatures,elemental composition of compounds and positive and negative ion mode mass spectrometry data. Result: A total of 26 compounds of cardiac glycoside were identified,including 25 compounds of cardiac glycoside from N. indicum host,and 5 compounds of cardiac glycoside from S. parasitic(parasitic on N. indicum). none of cardiac glycosides were found in S. parasitica (parasitic on O. fragrans ) and its O. fragrans host. Conclusion: It was rapid,accurate and comprehensive to identify cardiac glycosides in S. parasitica (parasitic on N. indicum) and its N. indicum host by UPLC-Q-TOF-MS/MS. S. parasitica itself does not contain cardiac glycosides,its host may impact the quality of S. parasitica by delivering cardiac glycosides, a kind of its characteristic compound.
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AIM To observe the effects of Taxilli Herba from six different hosts (Morus alba L.,Salix babylonica L.,Camellia oleifera Abel.,Castanea mollissima B1.,Liquidambar formosana Hance and Nerium indicum Mill.) on lowering blood pressure of spontaneously hypertensive rats (SHR).METHODS SHR were randomly divided into 14 groups,captopril positive group (20 mg/kg),model group,and Taxilli Herba groups of 6 different hosts,and each Taxilli Herba group was further divided into high-dose group (5.9 g/kg) and low-dose group (1.48 g/kg);WKY,in addition,was the blank control.And the 20-day consective correspondence medication was applied to the groups,each with eight rats.The caudal arterial systolic blood pressure (SBP) was measured by tail-cuff method before the administration,and on the 10th day and 20th day of the administration.Anaesthesia was performed at the blood collection 12 h after the last administration;and thus final serum contents of nitric oxide (NO) and changes of superoxide dismutase (SOD) activity,plasmatic contents of angiotensin Ⅱ (Ang Ⅱ) and endothelin-1 (ET-1) were determined.RESULTS From the data before and after administration,an SBP drop among all SHR groups was observed on the 10th day of administration,among which the blood pressure lowering effect by high-dose Taxilli Herba from Morus alba L.was very obvious (P ＜ 0.01);remarkable SBP decrease on the 20th day of administration induced by Taxilli Herba from Salix babylonica L,Liquidambarformosana Hance and Camellia oleifera Abel,and high-dose Taxilli Herba from Morus alba L,low-dose Taxilli Herba from Castanea mollissima B1 were detected (P ＜ 0.01).No significant SBP variation was available between the model group and Taxilli Herba groups after10-day administration;all the Taxilli Herba groups exhibited obvious effect in lowering SBP except Taxilli Herba from Nerium indicum Mill,low-dose Taxilli Herba from Morus alba L.and high-dose Taxilli Herba from Castanea mollissimaBl.after 20-day administration,compared to the model group (P ＜ 0.05).Highdose Taxilli Herba from Morus alba L.and Salix babylonica L significantly decreased plasmatic Ang Ⅱ and ET-1 contents of SHR groups in comparison with the model group (P ＜0.01).High-dose Taxilli Herba from Morus alba L.,Salix babylonica L.and Liquidambarformosana Hance significantly increased serum NO release and superoxide dismutase (SOD) activity (P ＜ 0.01).CONCLUSION Taxilli Herba from the five different hosts,except Nerium indicum Mill,can lower blood pressure,and there exists an effect difference due to the host variation.
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Objective To explore the expression level and possible reasons of Tau protein in Beagle dog model of ischemic cerebral white matter(WM) demylination.Methods Sixteen adult Beagle dogs were divided into the sham operation group(A) and observation group B,C and D according to the completely random mumber table method,4 cases in each group.Different.degrees of cerebral ischemia animal models were established by 4-vessel occlusion(4-VO) method.The bilateral ventricle edge white matter (WM) was selected.The oligodendrocyte precursors(OPCs) were labeled by NG2.The mature oligodendrocytes(Ols) were labeled by CNPase.Tau,NG2 and CNPase were detected by using the immunohistochemical method.The expression level was quantified by the mean optical value.The correlation among Tau,NG2 and CNGase was analyzed by adopting the Pearson linear correlation analysis.Results The HE staining showed obvious changes of WM demylination after chronic cerebral ischemic.The scores after LFB staining in the group A,B,C and D were(0.75 ± 0.71) points,(1.38 ± 0.06) points,(1.63 ± 0.52) points and (1.88 ± 0.64)points.Compared with the group A,the scores in the group B,C and D were much higher(P＜0.05).Compared with the group A,the expression levels of Tau protein and NG2 were significantly increased(P＜0.05),while the CNPase expression level was significantly decreased(P＜0.05).The Pearson correlation analysis showed that Tau expression level was positively correlated with the NG2 expression level(r=0.277,P=0.006);Tau and NG2 were negatively correlated with the CNPase level(r=-0.303,-0.402,P=0.003,0.001).Conclusion The increase of Tau expression in Beagle dog model of ischemic cerebral WM demylination may be related to the differentiation dysmaturity of OPCs.
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Objective To use high-performance liquid chromatography-time of flight mass spectrometry (HPLC-TOF/ MS) for examining the steroidal saponins in rat urine after oral administration of Paris polyphylla extract, so as to lay a foundation for studying the metabolism of steroidal saponins in vivo. Methods SD rats were administered with an oral dose of 1 6 g Paris polyphylla extracts/kg body weight. The urine samples were collected 24 h after administration by oral gavage. The sample analysis was carried out on a reverse phase MG-C18 column (3. 0 mm× 100 3. 0μm) using a gradient mobile phase system of acetonitrile-water containing 0. 1% formic acid. TOF/MS was applied for qualitative analysis under positive and negative ion modes. The steroidal saponins in rat urine were identified by using the accurate molecular weight obtained by TOF/ MS and formula database. Results A total of 20 steroidal saponins were identified in the rat urine, and two pairs of isomers were deduced through their fragment ions and standards: polyphyllin VI and pennogenin-3-0-α-L-rhamnopyranosyl(l →4)-α-L-rhamnopyranosyl (1 → 3)-[-α--rhamnopyranosyl (1 → 2)]-β-D-glucopyranoside; and gracillin and pennogenin-3-O-α-L-rhamnopyranosyKl → 4)-α-L-rhamnopyranosyl (1 → 4)-β-D-glucopyranoside. Conclusion The established analysis method is accurate, reliable for identifying steroidal saponins in vivo, which paves a way for further pharmacokinetics and pharmacodynamics study of Paris polyphylla.
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Silent information regulator 1 (SIRT1) is a NAD+-dependent histone deacetylase,which plays an important role in cerebral ischemia.It mainly stabilizes cerebral vascular endothelium,prevents vascular stenosis and the anti-inflammatory and anti-oxidative stress effects after cerebral ischemia.This article reviews the protective effects of STRT1 and its related mechanisms in cerebral ischemia.
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Recent studies have shown that Notch signaling pathway can improve neural stem cell selfrepair ability and promote angiogenesis,but at the same time,it will also aggravate the inflammatory response after cerebral ischemia.This article reviews the relationship between Notch signaling pathway and cerebral
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Objective To introduce an accelerated solvent extraction (ASE) -HPLC-TOF/MS method for simultaneous determination of Polyphyllin I, Polyphyllin II, Polyphyllin VI, Polyphyllin VII, Dioscin and Gracillin in Paris Polyphylla. Methods The extraction was performed by ASE under the following conditions: extracting solvent 70% ethanol; extracting temperature 120°C; pressure 1. 17 MPa (1 700 psi); and static time 6 min. The separation was carried out on a MGC18 column (3.0 mm× 100 mm, 3. 0 μm). Elution was done with a linear gradient mobile phase system consisting of 0. 1% formic acid (V/V) water and acetonitrile. The flow rate was set at 0. 8 ml/min, the sample injection volume was 3 μl, the column temperature was kept constant at 25°C, nebulizer gas pressure was 275. 8 kPa (40 psi), drying gas flow rate was 10 L/min, and gas temperature was 350°C. Results The calibration curves for Polyphyllin I, Polyphyllin II, Polyphyllin VI, Polyphyllin VII, Dioscin and Gracillin were linear within the range of 0. 500 0-50. 00 μg/ml (r = 0. 999 8), 0. 422 3-42. 23 μg/ml (r = 0. 999 7), 0. 612 0-61. 20 μg/ml (r = 0. 999 9), 0. 714 0-71. 40 μg/ml (r = 0. 999 5), 0. 448 0-44. 80 μg/ml (r = 0. 999 9) and 0. 436 0-43.60 μg/ml (r = 0. 9997), respectively; and the average recoveries (n=6) were 98. 9%, 98. 0%, 102. 5%, 101. 9%, 103. 1% and 97. 9%, respectively. Conclusion The introduced method is solvent-saving and time-saving; it can also be used for high-throughput analysis and determination of steroidal saponins in Paris Polyphylla.