Extraction and microencapsulation of bioactive compounds from pomegranate (Punica granatum var. Wonderful) residues

Pomegranate peels and seeds are industrial residues considered interesting sources of punicalagin (PU) and punicic acid (PA), respectively. To optimise their extraction process and protect them against environmental factors, pomegranate residues were extracted with supercritical CO₂ (SC‐CO₂) using a Box–Behnken design and then optimal extracts encapsulated by spray‐drying applying a 2² central composite design. Peel extracts showed a PU content of 0.4–9.5% with optimal extraction conditions of 400 bar, 43 °C and 20% ethanol. On the other hand, SC‐CO₂ seed extracts showed a PA content of 65.1–78.4% with 450 bar, 48 °C and 10% ethanol as optimal extraction parameters. Otherwise, the encapsulation efficiency of SC‐CO₂ extracts was significantly affected by core/wall material ratio and its quadratic effect. This parameter ranged from 35.1% to 72.4% for peel extracts and from 68.2% to 92.7% for seed extracts. Results showed that the proposed technologies are a feasible approach to the integral utilisation of residues from the pomegranate industry.     

安石榴苷对金黄色葡萄球菌毒力因子表达的抑制作用

分别采用溶血试验法、凝固酶效价法和酶联免疫反应法检测了安石榴苷处理后金黄色葡萄球菌溶血素、血浆凝固酶和总肠毒素的表达变化,并通过反转录实时荧光定量PCR技术检测毒力相关基因的表达情况。结果表明,安石榴苷能显著减少α-溶血素的产生,但对溶血素的抑制作用未呈现浓度依赖性;在高于1/4 MIC浓度下安石榴苷能降低血浆凝固酶的表达;在高于1/2 MIC浓度下,安石榴苷显著降低总肠毒素的产生;安石榴苷对肠毒素基因sea、溶血素基因hla和调节基因agrA的转录产生显著抑制作用,在1/8 MIC浓度下3个基因的相对表达量分别为对照组的3%,5%,5%。     

A Comparative Study Between Labeling and Reality: The Case of Phytochemical Composition of Commercial Pomegranate‐Based Products

Manufacturers are deeply involved in the development of new pomegranate‐based products, which have acquired great prestige due to many studies proving their potential health benefits. Commercial pomegranate products including capsules and supplements and juices and nectars were assayed. The contents of the key phytochemical compounds (punicalagin, ellagic acid, and total polyphenolic content) and the associated antioxidant capacity (DPPH^?, ABTS^?+, and FRAP) were analyzed. The experimental ranges of punicalagin and ellagic acid contents were 0.96 to 308 mg/g and 0.09 to 13.1 mg/g, respectively. Punicalagin content was positively correlated (P < 0.001) with DPPH^? and total polyphenolic content values. No significant (P > 0.05) correlation was observed among ellagic acid with the antioxidant capacity. The labeling standardization of these products is necessary due to the wide variability among “theoretically” similar pomegranate‐based products.     

安石榴苷与焦磷酸钠对肌原纤维蛋白氧化稳定性及凝胶性能的影响

研究添加安石榴苷、焦磷酸钠（sodium pyrophosphate,SPP）及其组合（P＋SPP）对肌原纤维蛋白（myofibrillar protein,MP）氧化稳定性和凝胶性能的影响。结果表明,安石榴苷和SPP添加均能有效抑制氧化诱导的蛋白羰基含量上升及自由氨基含量下降,且P＋SPP组合添加效果最佳;但所有添加剂均无法有效抑制氧化诱导的巯基含量降低。添加安石榴苷促进了氧化诱导的蛋白结构展开、显著抑制了蛋白粒径增大（抑制率为10.50%）,对于蛋白溶解度和热诱导凝胶过程中的流变学特性无显著影响,但明显降低了氧化MP凝胶的蒸煮损失和质构劣变。SPP添加同样促进了蛋白结构展开、抑制了蛋白粒径增大（抑制率为17.65%）、提高了氧化MP的溶解度（提高了9.78%）,显著改变了热诱导凝胶过程中MP的流变学行为,致使热诱导凝胶的蒸煮损失降低了42.56%,但明显降低了凝胶强度等质构指标。P＋SPP对MP凝胶性能的影响整体与SPP单独添加类似。因此,本研究抗氧化剂使用显著提高了MP的氧化稳定性及凝胶持水性,但凝胶的质构特性与抗氧化剂的种类密切相关。     

Antifungal Activity and DNA Topoisomerase Inhibition of Hydrolysable Tannins from Punica granatum L.

Punica granatum L. (pomegranate) fruit is known to be an important source of bioactive phenolic compounds belonging to hydrolysable tannins. Pomegranate extracts have shown antifungal activity, but the compounds responsible for this activity and their mechanism/s of action have not been completely elucidated up to now. The aim of the present study was the investigation of the inhibition ability of a selection of pomegranate phenolic compounds (i.e., punicalagin, punicalin, ellagic acid, gallic acid) on both plant and human fungal pathogens. In addition, the biological target of punicalagin was identified here for the first time. The antifungal activity of pomegranate phenolics was evaluated by means of Agar Disk Diffusion Assay and minimum inhibitory concentration (MIC) evaluation. A chemoinformatic analysis predicted for the first time topoisomerases I and II as potential biological targets of punicalagin, and this prediction was confirmed by in vitro inhibition assays. Concerning phytopathogens, all the tested compounds were effective, often similarly to the fungicide imazalil at the label dose. Particularly, punicalagin showed the lowest MIC for Alternaria alternata and Botrytis cinerea, whereas punicalin was the most active compound in terms of growth control extent. As for human pathogens, punicalagin was the most active compound among the tested ones against Candida albicans reference strains, as well as against the clinically isolates. UHPLC coupled with HRMS indicated that C. albicans, similarly to the phytopathogen Coniella granati, is able to hydrolyze both punicalagin and punicalin as a response to the fungal attack. Punicalagin showed a strong inhibitory activity, with IC₅₀ values of 9.0 and 4.6 µM against C. albicans topoisomerases I and II, respectively. Altogether, the results provide evidence that punicalagin is a valuable candidate to be further exploited as an antifungal agent in particular against human fungal infections.     

田口法优化石榴皮中3种活性成分的提取工艺

采用田口法优化乙醇回流提取石榴皮中3种酚酸——安石榴林、安石榴苷和鞣花酸,并使3种成分产率达到最大。通过单因素试验和田口法优化显著影响石榴皮提取的因素,包括乙醇体积分数、提取温度、提取时间、液料比、粒径和提取次数。结果表明,优化后取得的乙醇体积分数、提取温度、提取时间、液料比、粒径和提取次数分别为20%、70℃、2 h、50︰1(mL/g)、0.125 mm和提取2次。在这些最佳条件下,从1 g干燥的石榴皮粉末中可获得17.89±0.27 mg安石榴林、498.66±2.13 mg安石榴苷、7.30±0.05 mg鞣花酸,与预测值17.94 mg安石榴林、497.44 mg安石榴苷、7.26 mg鞣花酸符合。因此,达到利用田口法使安石榴林、安石榴苷和鞣花酸3种成分产率最大的目标。     

Punicalagin Mollifies Lead Acetate-Induced Oxidative Imbalance in Male Reproductive System

Punicalagin (PU) is a known antioxidant. The present study examined PU to protect against lead-induced oxidative stress (OS) testicular damage in mice. Significant increase in lipid peroxidation (LPO) after intraperitoneal injection of lead acetate (LA) indicated enormous generation of reactive oxygen species (ROS). Lead-induced OS has a direct effect on the differentiation of spermatogonial cells, showing a significant decline in sperm count. Supplementation of PU significantly changes values of LPO and glutathione (GSH) with a concomitant increase in sperm count, a marked decrease in the abnormal sperms, and a decline in the morphologically abnormal sperm population. Moreover, the histopathological evaluation of testes and epididymides showed severe changes in mice treated with LA. PU significantly induced nuclear factor erythroid-2 related factor 2-like 2 (Nrf2) expression and phase II enzymes, and data suggest that PU may inhibit OS through Nrf2 activation. The fertility test proved that PU might play an important role in male infertility treatment, especially in the type of infertility induced by OS.     

Mixed Pro- and Anti-Oxidative Effects of Pomegranate Polyphenols in Cultured Cells

In recent years, the number of scientific papers concerning pomegranate (Punica granatum L.) and its health properties has increased greatly, and there is great potential for the use of bioactive-rich pomegranate extracts as ingredients in functional foods and nutraceuticals. To translate this potential into effective strategies it is essential to further elucidate the mechanisms of the reported bioactivity. In this study HepG2 cells were supplemented with a pomegranate fruit extract or with the corresponding amount of pure punicalagin, and then subjected to an exogenous oxidative stress. Overall, upon the oxidative stress the gene expression and activity of the main antioxidant enzymes appeared reduced in supplemented cells, which were more prone to the detrimental effects than unsupplemented ones. No differences were detected between cells supplemented with the pomegranate juice or the pure punicalagin. Although further studies are needed due to the gaps existing between in vitro and in vivo studies, our results suggest caution in the administration of high concentrations of nutraceutical molecules, particularly when they are administered in concentrated form.