石榴叶提取物中鞣花酸在大鼠体内药物动力学研究

目的通过大鼠口服石榴叶提取物,研究其中活性成分鞣花酸的药动学特征。方法采用反相HPLC法,紫外检测,应用药动学软件3p87拟合大鼠体内鞣花酸的药动学模型。结果鞣花酸在大鼠体内呈二室模型分布,主要药动学参数为:α=1.07h-1,β=0.12h-1,Ka=5.67h-1,t1/2α=0.65h;t1/2β=5.94h;t1/2Ka=0.12h;K21=0.36h-1;K10=0.35h-1;K12=0.48h-1;AUC=0.85g·h·L-1。结论鞣花酸药动学特征为口服血药浓度低,大部分经胃吸收,达峰时间短,快速吸收、快速分布消除。石榴叶中鞣花酸的吸收大于鞣花酸单体的吸收。     

反相高效液相色谱法测定石榴皮中没食子酸的含量

目的建立石榴皮中没食子酸的含量测定方法。方法采用反相高效液相色谱法测定石榴皮中没食子酸的含量。色谱柱为C18柱,流动相为甲醇-0.05%磷酸溶液(5:95),流速0.8mL.min-1,检测波长273nm。结果没食子酸在0.0213～0.341mg.L-1范围内线性关系良好,r=0.9999;平均回收率为99.7%;RSD为0.6%。结论方法简便、快速,准确可靠,重复性好,可为石榴皮药材的质量控制提供参考。     

Preparation method and stability of ellagic acid-rich pomegranate fruit peel extract

A simple one-step purification using liquid–liquid extraction for preparing pomegranate peel extract rich in ellagic acid has been demonstrated. The method involved partitioning of the 10% v/v water in methanol extract of pomegranate peel between ethyl acetate and 2% aqueous acetic acid. This method was capable of increasing the ellagic acid content of the extract from 7.06% to 13.63% w/w. Moreover, the antioxidant activity of the extract evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay was also increased (ED₅₀ from 38.21 to 14.91?μg/mL). Stability evaluations of the ellagic acid-rich pomegranate peel extract in several conditions through a period of four months found that the extracts were stable either kept under light or protected from light. The extracts were also stable under 4° ± 2°C, 30° ± 2°C and accelerated conditions at 45°C with 75% relative humidity. However, study on the effect of pH on stability of the extract in the form of solution revealed that the extract was not stable in all tested pH (5.5, 7 and 8). These results indicated that the ellagic acid-rich pomegranate peel extract was stable when it was kept as dried powder, but it was not stable in any aqueous solution.     

In vitro evaluation of Spirulina platensis extract incorporated skin cream with its wound healing and antioxidant activities

Context: Algae have gained importance in cosmeceutical product development due to their beneficial effects on skin health and therapeutical value with bioactive compounds. Spirulina platensis Parachas (Phormidiaceae) is renowned as a potential source of high-value chemicals and recently used in skincare products.

Objective: This study develops and evaluates skin creams incorporated with bioactive S. platensis extract.

Materials and methods: Spirulina platensis was cultivated, the aqueous crude extract was prepared and in vitro cytotoxicity of S. platensis extract in the range of 0.001–1% concentrations for 1, 3 and 7?d on HS2 keratinocyte cells was determined. Crude extracts were incorporated in skin cream formulation at 0.01% (w/w) concentration and in vitro wound healing and genotoxicity studies were performed. Immunohistochemical staining was performed to determine the collagen activity.

Results: 0.1% S. platensis extract exhibited higher proliferation activity compared with the control group with 198% of cell viability after 3?d. Skin cream including 1.125% S. platensis crude extract showed enhanced wound healing effect on HS2 keratinocyte cell line and the highest HS2 cell viability % was obtained with this concentration. The micronucleus (MN) assay results indicated that S. platensis extract incorporated creams had no genotoxic effect on human peripheral blood cells. Immunohistochemical analysis showed that collagen 1 immunoreactivity was improved by increased extract concentration and it was strongly positive in cells treated with 1.125% extract incorporated skin cream.

Conclusions: The cell viability, wound healing activity and genotoxicity results showed that S. platensis incorporated skin cream could be of potential value in cosmeceutical and biomedical applications.     

Dietary supplementation of an ellagic acid-enriched pomegranate extract attenuates chronic colonic inflammation in rats

Dietary polyphenols present in Punica granatum (pomegranate), such as ellagitannins and ellagic acid (EA) have shown to exert anti-inflammatory and antioxidant properties. This study was designed to evaluate the effects of a dietary EA-enriched pomegranate extract (PE) in a murine chronic model of Cronh's disease (CD). Colonic injury was induced by intracolonic instillation of trinitrobenzensulfonic acid (TNBS). Rats were fed with different diets during 30 days before TNBS instillation and 2 weeks before killing: (i) standard, (ii) PE 250 mg/kg/day, (iii) PE 500 mg/kg/day, (iv) EA 10 mg/kg/day and (v) EA 10 mg/kg/day enriched-PE 250 mg/kg/day. Inflammation response was assessed by histology and MPO activity and TNF-α production. Besides, colonic expressions of iNOS, COX-2, p38, JNK, pERK1/2 MAPKs, IKBα and nuclear p65 NF-κB were studied by western blotting. MPO activity and the TNF-α levels were significantly reduced in dietary fed rats when compared with TNBS group. Similarly, PE and an EA-enriched PE diets drastically decreased COX-2 and iNOS overexpression, reduced MAPKs phosporylation and prevented the nuclear NF-κB translocation. Dietary supplementation of EA contributes in the beneficial effect of PE in this experimental colitis model and may be a novel therapeutic strategy to manage inflammatory bowel disease (IBD).     

新疆不同产地石榴花中总鞣质、没食子酸和鞣花酸的含量测定

目的测定新疆不同产地石榴花中总鞣质、没食子酸和鞣花酸的含量。方法采用紫外-可见分光光度法测定总鞣质的含量,HPLC法测定没食子酸和鞣花酸的含量。结果不同产地石榴花中总鞣质(38.44～113.98 mg·g~(-1))、没食子酸(2.39～3.81 mg·g~(-1))和鞣花酸(2.37～5.34 mg·g~(-1))含量差别均较大,和田皮山县石榴花中3种成分含量相对较高。结论该方法准确、专属性强,可用于石榴花药材的质量控制。     

Ameliorating effect of Celastrus paniculatus standardized extract and its fractions on 3-nitropropionic acid induced neuronal damage in rats: possible antioxidant mechanism

Context: Celastrus paniculatus Wild. (Celasteraceae) (CP) is a well-known Ayurvedic ‘Medhya Rasayana’ (nervine tonic), used extensively as a neuro-protective and memory enhancer, and in different central nervous system disorders.

Objective: To evaluate the effect of CP against 3-nitropropionic acid (3-NP) induced Huntington's disease (HD) like symptoms in Wistar male rats.

Materials and methods: The ethanol extract of CP seeds (CPEE), prepared by maceration, was standardized on the basis of linoleic acid content (6.42%) using thin layer chromatography densitometric analysis. Protective effect of CPEE (100 and 200?mg/kg) and its various fractions, viz., petroleum ether (40?mg/kg), ethyl acetate (2.5?mg/kg), n-butanol (7?mg/kg) and aqueous (18?mg/kg), administered orally for 20 days, against 3-NP (10?mg/kg, i.p. for 14 days) was assessed by their effect on body weight, locomotor activity, grip strength, gait pattern and cognitive dysfunction and biochemical parameters for oxidative damage in the striatum and cortex regions of the brain.

Results: CPEE (100 and 200?mg/kg) treated animals exhibited a significant (p?p?Conclusions: CPEE has a protective action against 3-NP induced HD like symptoms due to its strong antioxidant effect.     

Influence of ellagic acid on antioxidant defense system and lipid peroxidation in mice

Addition of ellagic acid (EA) to liver microsomes of mice resulted in a steady increase in inhibition of NADPH-dependent lipid peroxidation up to 2 mM concentration. The maximum of 70% inhibition of ascorbate-dependent lipid peroxidation was achieved at 1 mM concentration of EA. Feeding of EA significantly increased the levels of reduced glutathione and glutathione reductase in liver and lungs of male and female mice. However, there were no changes in the activities of catalase and superoxide dismutase. On the other hand, microsomes from liver and lungs of EA fed animals showed significantly suppressed NADPH- and ascorbate-dependent lipid peroxidation.     

Wound healing activity of alcoholic extract of Kaempferia galanga in Wistar rats

The wound healing effect of alcoholic extract of Kaempferia galanga (K. galanga) and its effect in dexamethasone suppressed wound healing was studied in Wistar rats. Three wound models viz. incision, excision and dead space wounds were used in this study. The parameters studied were breaking strength in case of incision wounds, epithelialization and wound contraction in case of excision wound and granulation tissue dry weight, breaking strength and hydroxyproline content in case of dead space wound. The dexamethasone treated group showed a significant (P < 0.001) reduction in the wound breaking strength when compared to control group in incision type of wound model. Coadministration of K. galanga with dexamethasone had significantly (P < 0.001) increased the breaking strength of dexamethasone treated group. In excision wound model, the percentage of the wound contraction was significantly (P < 0.05) increased by K. galanga only on 16th day and also it reversed the dexamethasone suppressed wound contraction on the 16 day. K. galanga significantly (P < 0.001) reduced the time required for epithelialization and reversed the epithelialization delaying effect of dexamethasone significantly (P < 0.001).     

Anti-inflammatory, analgesic and anti-oedematous effects of Lafoensia pacari extract and ellagic acid

Lafoensia pacari St. Hil. (Lythraceae) is used in traditional medicine to treat inflammation. Previously, we demonstrated the anti-inflammatory effect that the ethanolic extract of L. pacari has in Toxocara canis infection (a model of systemic eosinophilia). In this study, we tested the anti-inflammatory activity of the same L. pacari extract in mice injected intraperitoneally with beta-glucan present in fraction 1 (F1) of the Histoplasma capsulatum cell wall (a model of acute eosinophilic inflammation). We also determined the anti-oedematous, analgesic and anti-pyretic effects of L. pacari extract in carrageenan-induced paw oedema, acetic acid writhing and LPS-induced fever, respectively. L. pacari extract significantly inhibited leucocyte recruitment into the peritoneal cavity induced by beta-glucan. In addition, the L. pacari extract presented significant analgesic, anti-oedematous and anti-pyretic effects. Bioassay-guided fractionation of the L. pacari extract in the F1 model led us to identify ellagic acid. As did the extract, ellagic acid presented anti-inflammatory, anti-oedematous and analgesic effects. However, ellagic acid had no anti-pyretic effect, suggesting that other compounds present in the plant stem are responsible for this effect. Nevertheless, our results demonstrate potential therapeutic effects of L. pacari extract and ellagic acid, providing new prospects for the development of drugs to treat pain, oedema and inflammation.     

Matrix metalloproteinase-1 inhibitory activities of Morinda citrifolia seed extract and its constituents in UVA-irradiated human dermal fibroblasts

The objective of this study was to examine whether a 50% ethanolic extract (MCS-ext) of the seeds of Morinda citrifolia (noni) and its constituents have matrix metalloproteinase-1 (MMP-1) inhibitory activity in UVA-irradiated normal human dermal fibroblasts (NHDFs). The MCS-ext (10 μg/mL) inhibited MMP-1 secretion from UVA-irradiated NHDFs, without cytotoxic effects, at 48 h after UV exposure. The ethyl acetate-soluble fraction of MCS-ext was the most potent inhibitor of MMP-1 secretion. Among the constituents of the fraction, a lignan, 3,3'-bisdemethylpinoresinol (1), inhibited the MMP-1 secretion at a concentration of 0.3 μM without cytotoxic effects. Furthermore, 1 (0.3 μM) reduced the level of intracellular MMP-1 expression. Other constituents, namely americanin A (2), quercetin (3) and ursolic acid (4), were inactive. To elucidate inhibition mechanisms of MMP-1 expression and secretion, the effect of 1 on mitogen-activated protein kinases (MAPKs) phosphorylation was examined. Western blot analysis revealed that 1 (0.3 μM) reduced the phosphorylations of p38 and c-Jun-N-terminal kinase (JNK). These results suggested that 1 suppresses intracellular MMP-1 expression, and consequent secretion from UVA-irradiated NHDFs, by down-regulation of MAPKs phosphorylation.     

Effect of cocaine, 95% oxygen and ellagic acid on the development and antioxidant status of cultured rat embryos

Prenatal exposure to cocaine has been associated with adverse developmental effects and current data suggest cocaine induced malformations are caused by ischemic-reperfusion injury. This study was undertaken to assess a new in vitro model which uses a routine rat whole embryo culture system that incorporates a change in oxygen status, and to examine the effects of altered oxygen status and pretreatment with ellagic acid (EA), an anti-oxidant, after cocaine exposure. Embryos were evaluated by determining a developmental score and by measuring tissue reduced glutathione (GSH) levels. Following re-oxygenation with 95% O₂ for the last 6 h of culture, embryos treated with cocaine had reduced developmental scores and GSH levels. Embryos treated with cocaine and not re-oxygenated with 95% O₂ did not have reduced developmental scores. EA blocked the effects of cocaine on developmental score and GSH level. These data support ischemia-reperfusion injury as the mechanism of cocaine developmental toxicity.     

Comparison of antioxidant activities between salvianolic acid B and Ginkgo biloba extract （EGb 761）

AIM: To investigate and compare the antioxidant activities of salvianolic acid B (SalB) and Ginkgo biloba extract (EGb 761) in aqueous solution, rat microsomes and the cellular system. METHODS: Superoxide anion (O2-.) was generated using xanthine/xanthine oxidase system and phenazine methosulate/NADH system, and the effects of SalB and EGb 761 on the generation of O2-.were achieved by spectrophotometric measurement of the product formed on reduction of nitro blue tetrazolium. Two different methods were used to assess the scavenging effects of the extracts on hydroxyl radical (. OH): HPLC method was used for quantitation of . OH by oxy-radical trapping of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) to form DMPO-OH adducts in Fe2+-EDTA-H2O2 system. To confirm the HPLC data, .OH was also measured by spectrophotometry using a commercial detection kit. The anti-lipid peroxidation effects of the extracts in microsomes of rat brain, liver and kidney induced by ascorbate-NADPH were determined by thiobarbituric acid (TBA) method. The protective effects of the extracts on peroxide hydrogen (H2O2)-induced oxidative damage in SH-SY5Y cells were investigated by assessing cell viability assay, the level of lipid peroxidation, and the lactate dehydrogenase(LDH) release. RESULTS: Both SalB and EGb 761 were able to scavenge O2-. and . OH, inhibit lipid peroxidation of microsomes, and protect SH-SY5Y cells against H2O2-induced oxidative damage. However, the concentration of SalB was far lower than that of EGb 761 when a similar effect was obtained. CONCLUSION: The antioxidant efficiency of SalB was greater than that of EGb 761. These results suggest that SalB, like EGb 761, has promising potential in treating oxidative damage-derived neurodegenerative disorders.     

秀丽莓中鞣花酸的含量测定和抗氧化活性评测及对接研究

目的建立秀丽莓中鞣花酸的含量测定方法并对其抗氧化活性进行研究。方法采用RP-HPLC法,SunFireTMC18色谱柱(150mm×4.6mm,5μm),乙腈-4mL·L-1磷酸水(20∶80)为流动相;流速:1.0mL·min-1;检测波长:254nm;柱温:35℃。有机结合计算机分子模拟及体外DPPH·抗氧化活性评价方法对其可能的作用机制及活性进行深入探讨。结果鞣花酸的线性范围为0.081 92~0.409 60μg(r=0.999 7,n=6);平均回收率(n=6)为99.51%(RSD=2.00%)。该成分可通过结构中的酚羟基经氢键、苯环经π-π键作用与受体蛋白的氨基结合起效,具有较维生素C更强的抗氧化活性。结论该方法可迅速、准确地测定秀丽莓中的鞣花酸含量,可作为该药材中主要抗氧化成分鞣花酸质量控制和评价的有效方法,为该药材的进一步开发提供判断方法。     

Cardioprotective effect of whole fruit extract of pomegranate on doxorubicin-induced toxicity in rat

Context: Cardioprotective effects of various plants are generally attributed to their antioxidant activity. The whole fruit extract of pomegranate (WFEP), Punica granatum L. (Punicaceae), has a potent antioxidant activity.

Objective: To investigate cardioprotective effect of WFEP against doxorubicin (Dox)-induced cardiotoxicity in rats.

Materials and methods: Male Wistar rats were divided randomly into three groups of eight rats each: control (water, 5?mL/kg); Dox (10?mg/kg i.v.) and WFEP (100?mg/kg). Dox was administered in Dox and WFEP groups. After anesthetizing the animals on the last day, electrocardiogram was recorded and blood was analyzed for creatine kinase-MB isoenzyme (CK-MB), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) activities. Determinations of superoxide dismutase (SOD), reduced glutathione (GSH), lipid peroxidation (LPO) and histopathology of the heart tissues were carried out.

Results: The WFEP group showed decreased QT and increase in heart rate (p?<?0.05) compared to the Dox group. Significant decrease in CK-MB (p?<?0.01), LDH (p?<?0.05) and no such significant decrease in AST were observed as compared to the Dox group. There was significant increase in the level of GSH (p?<?0.05), whereas inhibition of LPO and increase in SOD concentration was not significant in the WFEP group compared to the Dox group. Histopathological study of the WFEP-treated group showed slight protection against myocardial toxicity induced by Dox.

Conclusion: Results indicate that WFEP has cardioprotective effect against Dox-induced cardiotoxicity in rats.     

石榴叶鞣质中短叶苏木酚在大鼠体内药代动力学变化

目的建立测定短叶苏木酚血药浓度的高效液相色谱方法,探讨大鼠ig石榴叶鞣质后体内短叶苏木酚动力学变化。方法色谱条件依利特Hypersil色谱柱;流动相为乙腈-5mmol·L-1磷酸二氢钾溶液(pH25;86∶14,V/V);流速10ml·min-1;紫外检测波长为276nm,以短叶苏木酚峰面积定量。结果短叶苏木酚在176~880μg·L-1内线性良好,回归方程为^Y=393E5X+182E3(n=5),相关系数r2=09994,血浆中最低检测浓度为176μg·L-1。其352,176,880μg·L-13个浓度的回收率>85%,日内日间变异<7%。大鼠ig石榴叶鞣质后短叶苏木酚体内吸收、分布及消除均较快,符合二室开放模型。结论短叶苏木酚可以部分的反映了石榴叶鞣质的药代动力学特征。     

Antioxidant,antibacterial, and anti-inflammatory activities of standardized brazilin-rich Caesalpinia sappan extract

Abstract

Context: Brazilin is a major active principle of Caesalpinia sappan L. (Leguminosae or Fabaceae). For industry aspects, brazilin-rich extract (BRE) has been prepared and standardized to contain 39% w/w brazilin. BRE may have more advantages than brazilin in term of a lower-cost production process.

Objectives: To investigate the antioxidant, antibacterial, and anti-inflammatory activities of BRE.

Material and methods: BRE was prepared by a simple one-step purification of the crude ethanol extract of C. sappan heartwood (CSE) using a Diaion® HP-20 column. The antioxidant activities were determined using three methods, including DPPH radical scavenging, reducing power, and β-carotene bleaching assays, at concentration ranges of 1–10, 10–100, and 10–100?µg/mL, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of BRE (15.6–1000?µg/mL) against Gram-positive and Gram-negative bacteria were determined by the broth microdilution method. Anti-inflammatory activity of BRE (0.1–5?µg/mL) was evaluated as anti-denaturation activity using bovine serum albumin as a substrate.

Results and discussion: On the basis of β-carotene bleaching assay, BRE showed antioxidant activity with an EC₅₀ value of 60.5?µg/mL, which was almost equal to that of pure brazilin (52.1?µg/mL). Gram-positive bacteria were more sensitive to all tested samples than Gram-negative bacteria. BRE possessed higher antibacterial activities than CSE, but lower than brazilin. MIC/MBC values of 62.5–125/125 and 250–500/250–500?µg/mL were obtained for BRE against Gram-positive and Gram-negative bacteria, respectively. A low concentration (0.1?µg/mL) of brazilin, BRE, and CSE showed anti-inflammatory activity by inhibiting protein denaturation up to 46.8, 54.1, and 61.9%, respectively.