Evaluation of an Undocumented Polyherbal (Faradin®) Used for the Treatment of Sickle Cell Disease in West Africa. Part I: Phytochemistry and Ex-vivo Anti-sickling Study

M. Adeyeye, D. Gbadero, L. Farayola, N. Olalere, Ravikiran Panakanti, Ibraheem Rajab, Sinni Moozhayil, C. Renault, Brandon Swinford, Magdalena Bujok
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引用次数: 3

Abstract

Background: An undocumented complementary and alternative medicine (CAM)Faradin® was screened for its phytochemicals, anti-sickling effect using a clinical protocol, and toxicity. Faradin is a polyherbal made up of Zanthoxylum zanthoxyloides, Alnus glutinosa and Alchornea cordofolia. Original Research Article Adeyeye et al.; BJPR, 17(1): 1-14, 2017; Article no.BJPR.33605 2 Methods: The polyherbal and the individual components (coded V, M, and F) were separately screened for phytochemicals such as alkaloids, phenols, flavonoids, sterols and coumarins, using standard tests. Institutional Review Board approved clinicalprotocolinvolving4 female de-identified sickle cell patients was used for the ex-vivo anti-sickling evaluation. The negative control was phosphate-buffered saline (PBS)-washed erythrocytes treated with 2% sodium metabisulfite (SMB) to cause a hypoxic state and result in HBSS polymerization or sickling. As positive control, 2% w/v p-hydroxy benzoic acid (PHBA) solution was used. The anti-sickling effect was studied by incubating the washed erythrocytes in various concentrations of Faradin Faradin: water v/v ratios (2.5:1 – 10:1) for 4 hours followed by further incubation with SMB for 11⁄2 hrs. The respective samples were examined under an optical microscope and the number of sickled cells counted. Cell viability to Faradin was also done using human primary hepatocytes (HPP) and Cell Titer-Glo Luminescent assay. Results: Faradin tested positive for flavonoids, phenols, coumarins, alkaloids and antioxidants. The V component tested positive for coumarins, alkaloids and antioxidants. The M component was positive for phenols, alkaloids and antioxidants while the F component tested positive for flavonoids, phenols and antioxidants. For the anti-sickling study, the negative control (SMB treated erythrocytes) showed 20-28% increase in sickling relative to the PBS-washed. The positive control caused a reversal of sickling by decreasing the percent sickled cells from 21% to 8%. There was a dose-dependent decrease in percent sickled cells; the highest dose reduced the percent sickled cells from 21% to 2%. The activity was likely due to the phenols, polyphenols, flavonoids and antioxidants in the CAM. Cell viability of Faradin® was greater than 85%. Conclusions: Thecombined phytochemicals in the polyherbal contributed to the reversal of sickling that is similar to the positive control. Faradin has a high potential for clinical effectiveness in the management of SCD.
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一种在西非用于治疗镰状细胞病的未证复方草药(Faradin®)的评价第一部分:植物化学和离体抗镰状细胞病研究
背景:一种未被证实的补充和替代药物(CAM)Faradin®对其植物化学成分、抗镰状细胞效应和毒性进行了筛选。Faradin是一种由花椒、Alnus glutinosa和Alchornea cordofolia组成的多草药。Adeyeye等;生物工程学报,17(1):1-14,2017;文章no.BJPR。方法:采用标准试验方法,分别对复方及单组分(编码V、M、F)进行生物碱、酚类、黄酮类、甾醇、香豆素等植物化学成分的筛选。机构审查委员会批准的临床方案涉及4名女性去识别镰状细胞患者,用于体外抗镰状细胞评估。阴性对照是磷酸盐缓冲盐水(PBS)冲洗的红细胞,用2%的代谢亚硫酸钠(SMB)处理,造成缺氧状态,导致HBSS聚合或镰状细胞。阳性对照采用2% w/v对羟基苯甲酸(PHBA)溶液。用不同浓度的法拉丁与水的v/v比(2.5:1 ~ 10:1)孵育冲洗后的红细胞4小时,再用SMB孵育11 / 2小时,研究其抗镰状细胞的作用。在光学显微镜下检查各自的样品并计算镰状细胞的数量。用人原代肝细胞(HPP)和细胞滴度-荧光荧光法测定细胞对Faradin的活力。结果:法拉丁的类黄酮、酚类、香豆素、生物碱和抗氧化剂检测呈阳性。V成分的香豆素、生物碱和抗氧化剂检测呈阳性。M组分对酚类、生物碱和抗氧化剂检测阳性,F组分对黄酮类、酚类和抗氧化剂检测阳性。对于抗镰状细胞的研究,阴性对照(SMB处理的红细胞)的镰状细胞比pbs洗涤的增加了20-28%。阳性对照通过将镰状细胞百分比从21%降低到8%而逆转镰状细胞。镰状细胞百分比呈剂量依赖性下降;最高剂量将镰状细胞百分比从21%减少到2%。这种活性可能与CAM中的酚类、多酚类、黄酮类和抗氧化剂有关。Faradin®的细胞存活率大于85%。结论:复方植物化学物质对镰刀病的逆转作用与阳性对照相似。Faradin在SCD治疗中具有很高的临床疗效潜力。
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