丹参四倍体优良品系有效成分的含量测定及药理学试验

运用HPLC法测定丹参 (SalviamiltiorrhizaBunge)同源四倍体优良品系 6 1 2 2 2及安徽、江苏和陕西产丹参不同部位、不同生长年限的有效成分 (3种丹参酮和 2种水溶性酚酸 )含量。丹参品系 6 1 2 2 2的有效成分含量明显高于其他产地丹参的含量 ;根中有效成分含量明显高于茎和叶的含量 ;丹参酮和丹参素的含量呈明显相关。初步药理实验结果表明 ,在相同剂量下 ,优良品系 6 1 2 2 2具有更好的药理活性。

Chemical shift assignments of two oleanane triterpenes from Euonymus hederaceus

1H-NMR and 13C-NMR assignments of 12-oleanene-3,11-dione (compound 1) were completely described for the first time through conventional 1D NMR and 2D shift-correlated NMR experiments using 1H-1HCOSY, HMQC, HMBC techniques. Based on its NMR data, the assignments of 28-hydroxyolean-12-ene-3,11-dione (compound 2) were partially revised.

Characterization and evaluation in vivo of baicalin-nanocrystals prepared by an ultrasonic-homogenization-fluid bed drying method

AIM： To improve the absorption and bioavailability of baicalin using a nanocrystal （or nanosuspension） drug delivery system. METHODS： A tandem, ultrasonic-homogenization-fluid bed drying technology was applied to prepare baicalin-nanocrystal dried powders, and the physicochemical properties of baicalin-nanocrystals were characterized by scanning electron microscopy, photon correlation spectroscopy, powder X-ray diffraction, physical stability, and solubility experiments. Furthermore, in situ intestine single-pass perfusion experiments and pharmacokinetics in rats were performed to make a comparison between the microcrystals of baicalin and pure baicalin in their absorption properties and bioavailability in vivo. RESULTS： The mean particle size of baicalin-nanocrystals was 236 nm, with a polydispersity index of 0.173, and a zeta potential value of-34.8 mV, which provided a guarantee for the stability of the reconstituted nanosuspension. X-Ray diffraction results indicated that the crystallinity of baicalin was decreased through the ultrasonic-homogenization process. Physical stability experiments showed that the prepared baicalin-nanocrystals were sufficiently stable. It was shown that the solubility of baicalin in the form of nanocrystals, at 495 ug·mL-1, was much higher than the baicalin-microcrystals and the physical mixture （135 and 86.4 ug·mL- 1, respectively）. In situ intestine perfusion experiments demonstrated a clear advantage in the dissolution and absorption characteristics for baicalin-nanocrystals compared to the other formulations. In addition, after oral administration to rats, the particle size decrease from the micron to nanometer range exhibited much higher in vivo bioavailability （with the AUC（0-t） value of 206.96 ± 21.23 and 127.95 ± 14.41 mg·L-1·h-1, respectively）. CONCLUSION： The nanocrystal drug delivery system using an ultrasonic-homogenization-fluid bed drying process is able to improve the absorption and in vivo bioavailability of baicalin, compared with pure baicalin coarse powder and micronized baicalin.

Structural characterization and identification of major constituents in Radix Scrophulariae by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry

AIM: To analyze the major constituents in Radix Scrophulariae(Scrophularia ningpoensis). METHOD: Radix Scrophulariae was analyzed by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry(ESI-Q-TOF MS/MS). Compounds were separated by HPLC using a C18 column and gradient elution of acetonitrile and 0.1 %(V/V) acetic acid-water. Negative ion mode was employed. RESULTS: A total of thirty-six compounds, including fourteen iridoid glycosides, nineteen phenylpropanoid glycosides, and three organic acids, were identified from Radix Scrophulariae based on the accurate mass measurement of precursor and product ions. Twenty-one of the constituents were identified by comparing their retention times(tR) and ESI-MS/MS data with those of reference standards and/or previous publications, while another fifteen compounds were tentatively identified or deduced according to their Q-TOF MS/MS data which afforded sufficient structural information. CONCLUSION: It is believed that this study is useful for the identification of constituents in Radix Scrophulariae, as well as related plants and complex prescriptions.

Chemical profiles and anticancer effects of saponin fractions of different polarity from the leaves of Panax notoginseng

AIM： To evaluate the chemical profiles and cytotoxic effects among the total saponin fraction （TSF）, 25% ethanol fraction （25EF）, 50% ethanol fraction （50EF）, and 85% ethanol fraction （85EF） prepared by macroporous resin from the leaves of Panax notoginseng. METHOD： The simultaneous determination of thirteen main saponins, as well as the chemical profiles of saponin fractions of different polarity, was made by HPLC-DAD and LC-ESI-MSn analysis. The cytotoxic effects were determined against KP4 cells （human pancreatic cancer）, NCI-H727 ceils （human lung cancer）, HepG2 cells （human hepatocellular cancer）, and SGC-7901 cells （human gastric adenocarcinoma）. RESULTS： Chemical analysis indicated that 85EF possessed the most abundant cytotoxic protopanaxadiol saponins, including the marker saponins F2, 20（R）-Rg3, 20（S）-Rg3, and Rh2. The MTT assay showed that 85EF also had the strongest cytotoxic effects among the four fractions. 25EF showed no anti-proliferative effects, while 50EF and TSF exhibited weak anti-proliferative activity. CONCLUSION： From the aspect of comprehensive utilization of resources, 85EF, enriched with low polarity PPD group saponins, is a new alternative source of anticancer saponins, and a promising botanical preparation for further anticancer studies.

Chemical constituents of Euphorbia tibetica and their biological activities

AIM: To investigate the chemical constituents and their biological activities of the aerial parts of Euphorbia tibetica. METHOD: Compounds were isolated and purified by various chromatographic methods, and their structures were elucidated through the use of extensive spectroscopic techniques including 2D-NMR, the structures of compounds 5 and 6 were confirmed by single-crystal X-ray crystallography. Bioactivities of all the isolated compounds were evaluated by the MTT method on A549 and anti-angiogenesis assay. RESULTS: One new compound, methyl 4-epi-shikimate-3-O-gallate(1), together with twenty-three known constituents(2–24) were isolated from the aerial parts of E. tibetica. CONCLUSION: Compound 1 is new, and the other compounds were isolated from this plant for the first time. Compounds 5–7, 9, 11, 17, 18 and 20 exhibited inhibitory effects on the growth of human lung cancer cell A549 and compounds 5, 7, 12, 13, 17 and 19 showed anti-angiogenic effects in a zebrafish model.

Ursolic acid induces U937 cells differentiation by PI3K/Akt pathway activation

AIM: Ursolic acid(UA), a pentacyclic triterpenoid, is used as an anti-inflammatory and anti-cancer agent. There were few studies on the effects of UA on differentiation, and this is the first time to elucidate the potential effect and molecular mechanism of UA on inducing differentiation in the human leukemia cell line U937. METHODS: Wright-Giemsa staining, nitroblue tetrazolium reduction assay and flow cytometric analysis were utilized to demonstrate the differentiation of U937 cells induced by UA. Western blotting and immunofluorescence assay were used to investigate the possible mechanism. RESULTS: It was found that UA could induce the differentiation of U937cells and Akt-activity was significantly increased during differentiation. Additionally, LY294002, a PI3K-Akt inhibitor, could block the differentiation of U937 cells induced by UA. CONCLUSION: UA could induce the differentiation of U937 cells by activating the PI3K/Akt pathway, and it could be a potential candidate as a differentiation-inducing agent for the therapy of leukemia.

DT-13, a saponin of dwarf lilyturf tuber, exhibits anti-cancer activity by down-regulating C-C chemokine receptor type 5 and vascular endothelial growth factor in MDA-MB-435 cells

AIM： To investigate the anticancer activity of DT-13 under normoxia and determine the underlying mechanisms of action. METHODS： MDA-MB-435 cell proliferation, migration, and adhesion were performed to assess the anticancer activity of DT-13, a saponin from Ophiopogonjaponicus, in vitro. In addition, the effects of DT-13 on tumor growth and metastasis in vivo were evaluated by orthotopic implantation of MDA-MB-435 cells into nude mice; mRNA levels of vascular endothelial growth factor （VEGF）, C-C chemokine receptor type 5 （CCR5） and hypoxia-inducible factor 1a （HIF-1a） were evaluated by real-time quantitative PCR; and CCR5 protein levels were detected by Western blot assay. RESULTS： At 0.01 to 1 umol·L -1, DT-13 inhibited MDA-MB-435 cell proliferation, migration, and adhesion significantly in vitro. DT-13 reduced VEGF and CCR5 mRNAs, and decreased CCR5 protein expression by down-regulating HIF-1 a. In addition, DT-13 inhibited MDA-MB-435 cell lung metastasis, and restricted tumor growth slightly in vivo. CONCLUSION： DT-13 inhibited MDA-MB-435 cell proliferation, adhesion, and migration in vitro, and lung metastasis in vivo by reducing VEGF, CCR5, and HIF-la expression.

Pyrolysis characteristics of Radix Rhizoma Rhei, Cortex Moudan Radicis, and Radix Sanguisorbae and correlations with the carbonizing process of Chinese herbs

AIM： The aim of the work is to study the pyrolysis characteristics of Radix Rhizoma Rhei, Cortex Moudan Radicis, and Radix Sanguisorbae in an inert atmosphere of argon （Ar）, and to investigate the mechanism of the carbonizing process of the three traditional Chinese herbs. METHODS： The pyrolysis characteristics of the crude materials and their extracts were studied by thermogravimetry-mass spectrometry （TG-MS） in a carrier gas of argon, coupled with Fourier transform infrared spectrometry （FTIR） and scanning electron microscopy （SEM） methods. Correlation of the pyrolysis behaviors with the carbonizing process by stir-frying of traditional Chinese medicines was made. RESULTS： Within the temperature range of 200-300 ℃, which is the testing range for the study of the carbonizing process of Chinese herbs, the temperatures indicated by the maximum weight loss rate peak of the above three extracts were taken as the upper-limit temperatures of the carbonizing process of the herbs, and which were 200, 240 and 247 ℃ for Radix Rhizoma Rhei, Cortex Moudan Radicis, and Radix Sanguisorbae, respectively. The ion monitoring signal peaks detected by the TG-MS method corresponded with reports that the level of chemical components of traditional Chinese medicinal materials would decrease after the carbonizing process. It was confirmed by Fourier transform infrared spectrometry （FTIR） and scanning electron microscopy （SEM） methods that better results of ＂medicinal property preservation＂ could be obtained by heating at 200 ℃ for Radix Rhizoma Rhei, at about 250 ℃ for Cortex Moudan Radicis, and Radix Sanguisorbae, as the relative intensity values of the common peaks were among the middle of their three carbonized samples by programmed heating. CONCLUSION： The upper-limit temperatures of the carbonizing process for Radix Rhizoma Rhei, Cortex Moudan Radicis and Radix Sanguisorbae were 200, 240 and 247 ℃ respectively. It is feasible to research the mechanism and technology of the carbonizing process of traditional Chinese medicinal materials using thermogravimetry, Fourier transform infrared spectrometry, and scanning electron microscopy methods.

Crebanine inhibits voltage-dependent Na～+ current in guinea-pig ventricular myocytes

AIM: To study the effects of crebanine on voltage-gated Na+ channels in cardiac tissues. METHODS: Single ventricular myocytes were enzymatically dissociated from adult guinea-pig heart. Voltage-dependent Na+ current was recorded using the whole cell voltage-clamp technique. RESULTS: Crebanine reversibly inhibited Na+ current with an IC50 value of 0.283 mmol?L-1(95% confidence range: 0.248-0.318 mmol?L-1). Crebanine at 0.262 mmol?L-1 caused a negative shift(about 12 mV) in the voltage-dependence of steady-state inactivation of Na+ current, and retarded its recovery from inactivation, but did not affect its activation curve. CONCLUSION: In addition to blocking other voltage-gated ion channels, crebanine blocked Na+ channels in guinea-pig ventricular myocytes. Crebanine acted as an inactivation stabilizer of Na+ channels in cardiac tissues.

Potent anti-angiogenicactivity of B19—a mono-carbonyl analogue of curcumin

AIM： The compound B19 （C21H22O5） is a newly synthesized, mono-carbonyl analog of curcumin that has exhibited potentialantitumor effects. This present study was performed to identify the anti-angiogenic activity of this compound. METHODS AND RESULTS： B19 inhibited migration and tube formation of human umbilical vein endothelial cells, and arrested microvessel outgrowth from rat aortic rings.ln addition, B19 suppressed the neovascularization of chicken chorioallantoic membrane. Mechanistic studies revealed that B19 suppressed the downstream protein kinase activation of vascular endothelial growth factor （VEGF） by decreasing phosphorylated forms of serine/threonine kinase Akt, extracellular signal-regulated kinase, and p38 mitogen-activated protein kinase, with or without stimulating vascular endothelial growth factor （VEGF）. CONCLUSIONS： B 19 exerted anti-angiogenic activity in vitro and ex vivo, which suggests that it merits further investigation as a promising anticancer angiogenesis compound.

Ethnopharmacology, phytochemistry and pharmacology of Tephrosia purpurea

Tephrosia purpurea(L.) Pers. is popularly known as ‘Sarapunkha' in classical Ayurvedic texts. It is a perennial plant belonging to the family Fabaceae, and occurs throughout the Indian subcontinent. T. purpurea is traditionally used to treat spleenomegaly, cirrhosis, cough and cold, abdominal swelling and as an antidote in the Ayurvedic system of medicine. Phytochemical investigations indicate the presence of semiglabrin, pongamole, lanceolatins A and B, rutin, lupeol, and β-sitosterol. Flavonoids including(+)-tephrorin A and B,(+)-tephrosone, an isoflavone, 7, 4'-dihydroxy-3', 5'-dimethoxyisoflavone and a chalcone,(+)-tephropurpurin were isolated from the whole plant. Pharmacological activities of different parts of the plant reported include anti-inflammatory, antiulcer, antimicrobial, antioxidant, antiallergic, antidiabetic, hepatoprotective, antitumor and insect repellent activity. In the present review, the literature on the phytochemical and pharmacological investigations of Tephrosia purpurea(L.) Pers. are summarized to August, 2012.

Triterpenoid saponins from Patrinia scabra

AIM： To study the chemical constituents and bioactivity of the roots ofPatrinia scabra Bunge. METHODS： The chemical constituents were isolated using various chromatographic methods, and the structures were elucidated on the basis of spectral analysis and chemical methods. In addition, cytotoxic activities toward HepG2 cells were tested by the MTT method. RESULTS： A new triterpenoid saponin, 3-O-（4＇-isovaleryl）-O-β-D-xylose-12,30-dihydroxy-oleanane-28,13-1actone-22-O- β-D-glucoside （1）, along with two known triterpenoid saponins, acanthopanax saponin CP3 （2） and foetoside C （3）, were isolated. CONCLUSION： The aglycone of compound 1 was a new skeleton derivative of oleanolic acid. Compound 2 showed strong cyto- toxicity to HePG2 cells （IC50 1.49 umol.L-1）.

Mixed nanomicelles loaded with thymopeptides-sodium deoxycholate/phospholipid improve drug absorption

AIM: To improve the absorption of thymopeptides(TH) by preparing sodium deoxycholate/phospholipid-mixed nanomicelles(SDC/PL-MMs). METHODS: TH-SDC/PL-MMs were prepared by a film dispersion method, and then evaluated using photon correlation spectroscopy(PCS), zeta potential measurement, as well as their physical stability after storage for several days. Furthermore, in situ intestinal single-pass perfusion experiments and pharmacodynamics in immunodeficient mice were performed to make a comparison with TH powders and the control drug in absorption properties. RESULTS: A narrow size distribution of nanomicelles, with a mean particle size of(149 ± 8.32) nm and a zeta potential of(-31.05 ± 2.52) mV, was obtained. The in situ intestine perfusion experiments demonstrated a significant advantage in absorption characteristics for TH compared to the other formulations, and oral administration of TH-SDC/PL-MMs potentiated an equivalent effect with i.h. TH in pharmacodynamic studies in immunodeficient mice. CONCLUSIONS: TH-SDC/PL-MMs prepared by a film dispersion method are able to improve the absorption of TH. SDC/PL-MMs might be a good approach for the more effective delivery of drugs like TH.