Quantitative determination of four compounds and fingerprint analysis in the rhizomes of Drynaria fortunei (Kunze) J. Sm.

A rapid, sensitive, and accurate reversed-phase high-performance liquid chromatography with photodiode array detection method was developed for both quantitative determination of four compounds (caffeic acid-4-O-β-d-glucopyranoside, 5,7-dihydroxychromone-7-O-rutinoside, neoeriocitrin and naringin) and fingerprint analysis of the rhizomes of Drynaria fortunei (Kunze) J. Sm. The chromatographic separation was accomplished on an MZ-C18 column (4.6 × 250 mm, 5 μm) using gradient elution with acetonitrile and 0.02% aqueous acetic acid, at a flow rate of 1.0 mL min⁻¹, an operating temperature of 25°C, and a wavelength of 260 nm. The four compounds showed good regression relationship (R ² > 0.9990) within linear ranges, and their recoveries were in the range of 98.11–102.23%. In the chromatographic fingerprint, thirteen common peaks were found and selected as characteristic peaks to assess the consistency of ten batches of the rhizomes of D. fortunei. The results indicate that the method of multiple compounds determination in combination with chromatographic fingerprint analysis is suitable for systematic quality evaluation of D. fortunei.     

HPLC法测定骨碎补酊中柚皮苷含量

目的建立骨碎补酊的质量标准。方法采用HPLC法测定柚皮苷含量。结果柚皮苷在9.76~146.4ng范围内呈良好线性关系,平均加样回收率为98.9%,RSD为1.5%。结论本法可用于骨碎补酊的质量控制。     

Osteoblastic proliferation stimulating activity of Psoralea corylifolia extracts and two of its flavonoids.

Psoralea corylifolia L. fruit extracts exhibited osteoblastic proliferation stimulating activity in UMR106 cell line cultured in vitro. The flavonoids of corylin and bavachin were isolated and identified as active principles by activity-guided fractionation. The results suggested that Psoralea corylifolia L. fruit extracts and corylin and bavachin might stimulate bone formation or have potential activity against osteoporosis.     

Effect of Guizhi Fuling capsule and its extracts on human breast cancer cells proliferation

OBJECTIVE To evaluate the effect of Guizhi Fuling Capsule active pharmaceutical ingredient(API)and its fractions on human breast cancer cells proliferation by high-throughput screening assay.METHODS The crude fractions were obtained from the extraction and elution of the API of Guizhi Fuling Capsule,and 929 standard fractions were obtained by the optimal separation conditions.Sulforhodamine B(SRB)method was used to evaluate the effects of the Guizhi Fuling capsule API and929 kinds of fractions on the proliferation of human breast cancer cells MCF-7 and MDA-MB-231.RESULTS The Guizhi Fuling capsule API had a strong ability to inhibit the proliferation of MCF-7 cells at high concentration and the ability to inhibit MDA-MB-231 cells' proliferate at low concentration following 72 h treatment;some samples of 929 fractions(5μg·mL~(-1))was found to have a breast cancer cell growth inhibition rate above 50%,without toxicity on HUVECs proliferation.CONCLUSION The API of Guizhi Fuling capsule had significant cytotoxicity effects on these two human breast cancer cells,with significant concentration-and time-dependent manner.     

狗脊化学成分的分离与鉴定

对狗脊Cibotium barometz(L.)J.Sm的干燥根茎的氯仿提取物及正丁醇提取物经硅胶柱色谱分离,得到了5个化合物,它们分别是金粉蕨素（I.onitin),原儿茶酸（Ⅱ,protocatechuric acid),β－谷甾醇（Ⅳ,β－sitosterol),胡萝卜苷（Ⅴ,daucosterol),5－羟甲糠醛（Ⅲ,2－furancarboxaldehydl-50-hydroxymethyl),其中化合物（Ⅲ）为首次从该属植物中分离得到。     

Platelet-activating factor (PAF) receptor binding antagonist activity of the methanol extracts and isolated flavonoids from Chromolaena odorata (L.) King and Robinson

The leaf, stem and root extracts of Chromolaena odorata were evaluated for their effect on platelet-activating factor (PAF) receptor binding on rabbit platelets using 3H-PAF as a ligand. The leaf extract demonstrated high PAF receptor binding inhibitory activity of 79.2+/-2.1% at 18.2 microg/ml. A total of eleven flavonoids were subsequently isolated from the active leaf extract and evaluated for their effects on PAF receptor binding. Eight of the flavonoids exhibited >50% inhibition on the binding activity at 18.2 microg/ml. These flavonoids were identified as eriodictyol 7,4'-dimethyl ether, quercetin 7,4'-methyl ether, naringenin 4'-methyl ether, kaempferol 4'-methyl ether, kaempferol 3-O-rutinoside, taxifolin 4'-methyl ether, taxifolin 7-methyl ether and quercetin 4'-methyl ether. Their IC50 values ranged from 19.5 to 62.1 microM.     

L-carnitine and taurine synergistically inhibit the proliferation and osteoblastic differentiation of vascular smooth muscle cells

Aim:

To investigate the synergistic action of L-carnitine (LC) and taurine (TAU) on the proliferation and osteoblastic differentiation of vascular smooth muscle cells (VSMCs).

Methods:

DNA and protein synthesis of VSMCs were assessed using scintillation counting. Alkaline phosphatase (ALP) activity and calcium content were determined to investigate the effects of LC and TAU on the osteoblastic differentiation and mineralization of VSMCs. TAU uptake by VSMCs was assayed. RNA interference was used to down-regulate the expression of the TAU transporter (TAUT) in rat VSMCs.

Results:

LC and TAU synergistically inhibited the proliferation and β-glycerophosphate (β-GP)-induced osteoblastic differentiation of VSMCs as evidenced by the decreased [³H]thymidine incorporation, ALP activity and calcium deposition. Furthermore, LC stimulated the TAU uptake and TAUT expression in VSMCs. Suppression of TAUT with short hairpin RNA (shRNA) abolished the synergistic action of LC and TAU in VSMCs.

Conclusion:

The synergistic inhibitory action of LC and TAU on the proliferation and osteoblastic differentiation of VSMCs is attributable to the up-regulation of TAUT expression and TAU uptake by LC.     

克罗米酚对骨髓间质干细胞增殖和向成骨细胞分化的影响

目的　在离体条件下研究克罗米酚(CLO)对小鼠骨髓间质干细胞(BMSCs)增殖和向成骨细胞分化的影响。方法　在含经活性炭吸附的 10%血清的无酚红α MEM中加入β 磷酸甘油和维生素C诱导小鼠BMSCs向成骨细胞分化,同时加入 0 1nmol·L-1至 10nmol·L-1CLO处理细胞。用细胞计数来反映细胞增殖情况;测定碱性磷酸酶 (ALP)活性与钙的沉积量反映细胞向成骨细胞分化状态;用试剂盒来检测一氧化氮 (NO)的产物。结果　CLO ( 0 1 ~10nmol·L-1 )呈剂量依赖性地增加小鼠BMSCs的数目,ALP活性和钙的沉积量,同时培养基中NO代谢产物明显增加。分别加入雌激素受体 (ER)拮抗剂ICI182, 780 (0 1μmol·L-1 )及一氧化氮合酶抑制剂L NAME( 6mmol·L-1 )均可阻止CLO促进BMSCs的增殖及向成骨细胞分化的作用,并取消NO代谢产物的增加。结论　CLO在 0 1 ~10nmol·L-1剂量范围内具有雌激素样受体激活效应,可通过ER/NO途径促进小鼠骨髓间质干细胞的增殖及向成骨细胞分化。     

K(Ca) channel-opening activity of Ginkgo Biloba extracts and ginsenosides in cultured endothelial cells

1. Extracts of Ginkgo biloba (EGb) and ginsenosides (GS) have been reported to induce vasorelaxation. In the present study, the role of K+ channels in the action of EGb and GS to activate nitric oxide synthase (NOS) activity was investigated in cultured endothelial cells. 2. Nitric oxide synthase activity of cultured endothelial cells detected by the reduced nicotinamide adenine dinucleotide phosphate (NADPH) histochemistry method was significantly increased after treatment with 20 microg/mL EGb or 40 microg/mL GS plus 10 mmol/L L-arginine. The effect was completely abolished by the addition of 0.5 micromol/L Nomega-nitro-L-arginine, an inhibitor of NOS, to the incubation medium and partially inhibited by 10 micromol/L tetraethylammonium (TEA), an inhibitor of Ca2+-activated K+ (KCa) channels. 3. Application of EGb to the intracellular surface of excised inside-out patches activated K+ channels in a concentration-dependent manner in the concentration range 1-100 microg/mL. Channel activity was also activated by application of GS at concentrations ranging from 1 to 300 microg/mL. The modulation of channel activity was inhibited by 0.5 mmol/L TEA but not by 0.5 mmol/L glibenclamide, an inhibitor of ATP-sensitive K+ channels. 4. Thus, in cultured endothelial cells, the increase in NOS activity induced by EGb or GS depends on the activity of KCa channels. These compounds may regulate nitric oxide release by changing the cell membrane potential in vascular endothelial cells.     

Inhibitory effect of murine kidney extracts on the proliferation of murine mast cells

We examined the effect of murine kidney extract (MKE) on the clonal growth of mast cells from murine peritoneal cells. Adding MKE resulted in a 40% inhibition of colony formation of mast cells in a methylcellulose culture, and a 90% decrease in mast cell numbers and histamine content in mast cells in a liquid culture containing stem cell factor and interleukin-3. The mast cell inhibitory factors in MKE were heat sensitive proteins of approximately 560 and 24 kDa. These results suggest that MKE contains regulators that suppress the growth of murine mast cells and histamine synthesis.     

The biphasic effects of cyclopentenone prostaglandins, prostaglandin J(2) and 15-deoxy-Delta(12,14)-prostaglandin J(2) on proliferation and apoptosis in rat basophilic leukemia (RBL-2H3) cells

Mast cells produce chemical mediators, including histamine and arachidonate metabolites such as prostaglandin D(2) (PGD(2)) after antigen stimulation. Cyclopentenone prostaglandins of the J series, prostaglandin J(2) (PGJ(2)) and 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), are thought to be derivatives of PGD(2). In this study, the biphasic effects of the PGJ(2) and 15d-PGJ(2) on proliferation and apoptosis in rat basophilic leukemia cells (RBL-2H3), a tumor analog of mast cells, were examined. At low concentrations, 1 or 3 microM PGJ(2) and 15d-PGJ(2) induced cell proliferation, respectively. At high concentrations (10-30 microM) both the inhibition of viability and decrease in histamine content in RBL-2H3 cells were dose dependent. These effects were independent of the nuclear hormone receptor, peroxisome proliferator-activated receptor gamma (PPARgamma), since troglitazone, an agonist of PPARgamma did not cause any effects in RBL-2H3 cells. Cell death induced by PGJ(2) and 15d-PGJ(2) was the result of apoptotic processes, since RBL-2H3 cells treated with 30 microM of the prostaglandins had condensed nuclei, DNA fragmentation and increase in activities of caspase-3 and -9. Moreover, PGJ(2) or 15d-PGJ(2)-induced apoptotic effects were prevented by the caspase inhibitor, z-VAD-fmk. In conclusion, the PGJ(2) or 15d-PGJ(2)-induced apoptosis in RBL-2H3 cells occurs mainly via mitochondrial pathways instead of by PPARgamma-dependent mechanisms.     

Effect of beta-alanyl-L-histidinato zinc on osteoblastic MC3T3-E1 cells: increases in alkaline phosphatase and proliferation.

The effect of beta-alanyl-L-histidinato zinc (AHZ) on bone metabolism was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37 degrees C in a CO2 incubator in plastic dishes containing alpha-modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus various concentrations of AHZ or zinc sulfate, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10(-6)-10(-4) mol/l) stimulated proliferation of cells. AHZ increased alkaline phosphatase activity in a dose-related manner up to 10(-5) mol/l; the increase was about 2-fold over the control value. Studies on the effect of actinomycin D or cycloheximide treatment indicated that AHZ may enhance de novo synthesis of the enzyme. AHZ also increased deoxyribonucleic acid (DNA) content dose dependently (10(-6)-10(-4) mol/l). This increase was completely blocked by treatment with cycloheximide. The AHZ (10(-5) mol/l)-induced increases in alkaline phosphatase activity and DNA content were entirely abolished by the presence of dipicolinate (10(-4) mol/l), a chelator of zinc, indicating that the effect of AHZ needs zinc. However, AHZ had a potent effect, more than that of zinc sulfate, on alkaline phosphatase activity and DNA content. The present results indicate that AHZ has a direct specific anabolic effect on osteoblastic cells in vitro and that this effect is related to protein synthesis.     

The activity of flavonoids extracted from Tanacetum microphyllum DC. (Compositae) on soybean lipoxygenase and prostaglandin synthetase

• 1.1. The in vitro effects of centaureidin and 5,3′-dihydroxy-4′-methoxy-7-carbomethoxyflavonol (Fig. 1), two anti-inflammatory flavonoids extracted from Tanacetum microphyllum DC., have been examined on both cyclooxygenase and lipoxygenase activity.
• 2.2. These flavonoids produced an inhibition of soybean lipoxygenase activity in a dose-dependent manner, with IC₅₀ values (20 and 29 μM respectively) similar to the reference drug.
• 3.3. The IC₅₀ values for the in vitro inhibition of cyclooxygenase activity by these flavonoids, were higher than those that produced lipoxygenase activity (318 and 60μM respectively).
• 4.4. These results suggest that the anti-inflammatory activity of our flavonoids may, at least in part, be due to the inhibition of leukotriene synthesis.
• 5.5. This is the first report of the biological activity in vitro of these compounds.

     

淫羊藿总黄酮及其含药血清对成骨细胞增殖及功能表达的影响

目的研究淫羊藿总黄酮及其含药血清对体外培养大鼠成骨细胞(ROB)增殖和功能表达的影响。方法将淫羊藿总黄酮(TFE)以0.2、2、20、100、200 mg.L-15种质量浓度、含淫羊藿总黄酮大鼠血清(SRAT)以2%、4%、8%、16%等4种体积浓度分别加入新生大鼠颅骨成骨细胞培养液中,研究其对细胞增殖和功能表达的影响。细胞增殖采用MTT法进行分析,细胞功能表达是于培养d 5、10、15、20分别检测碱性磷酸酶活性(比色法)、骨钙素分泌量(放射免疫法)、钙盐沉积量(比色法)和矿化结节(组织化学法)等。结果TFE5种质量浓度均对ROB细胞增殖和分化无影响,2%和4%SRAT则表现出强烈的刺激细胞增殖活性并提高碱性磷酸酶活性、骨钙素分泌量、矿化结节数和钙盐沉积量。结论淫羊藿治疗骨质疏松的直接物质基础可能并非TFE,而与其经口给予后吸收入血的TFE代谢产物及其诱生物有关。血清中的TFE代谢产物可促进细胞的功能表达增殖。