Institute of Pharmaceutical Biology and Phytochemistry, University of Münster, Münster, Germany.
Phytomedicine. 2011 May 15;18(7):617-24. doi: 10.1016/j.phymed.2010.08.020. Epub 2010 Oct 30.
Leaves from Phyllanthus muellerianus (Kuntze) Exell. are traditionally used for wound healing in Western Africa. Aqueous extracts of dried leaves recently have been shown to stimulate proliferation of human keratinocytes and dermal fibroblasts. Within bioassay-guided fractionation the ellagitannins geraniin (1), corilagin (2), furosin (3), the flavonoids quercetin-3-O-β-D-glucoside (isoquercitrin), kaempferol-3-O-β-D-glucoside (astragalin), quercetin-3-O-D-rutinoside (rutin), gallic acid, methyl gallate, caffeic acid, chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeoylmalic acid (phaselic acid) have been identified in P. muellerianus for the first time. Geraniin was shown to be the dominant component of an aqueous extract. Suitable analytical methods for quality control of geraniin in P. muellerianus extract (methanol/water, 70/30) have been developed and validated based on ICH guidelines (ICH-compliant protocol). Geraniin and furosin increased the cellular energy status of human skin cells (dermal fibroblasts NHDF, HaCaT keratinocytes), triggering the cells towards higher proliferation rates, with fibroblasts being more sensitive than keratinocytes. Highest stimulation of NHDF by geraniin was found at 5 μM, and of keratinocytes at 50-100 μM. Furosin stimulated NHDF at about 50 μM, keratinocytes at about 150-200 μM. Necrotic cytotoxicity of geraniin, as measured by LDH release, was observed at 20 μM for NHDF and 150 μM for keratinocytes. Toxicity of furosin--less than that of geraniin--was observed at > 400 μM. Furosin and geraniin stimulated the biosynthesis of collagen from NHDF at 50 μM and 5-10 μM respectively. Geraniin at 105 μM significantly stimulated the differentiation in NHEK while furosin had a minor influence on the expression of involucrin and cytokeratins K1 and K10. The study proves clearly that hydrophilic extracts from P. muellerianus and especially the lead compound geraniin exhibit stimulating activity on dermal fibroblasts and keratinocytes, leading to increased cell proliferation, barrier formation and formation of extracellular matrix proteins. From these findings the traditional clinical use of such extracts for wound healing seems to be justified.
叶从 Phyllanthus muellerianus（Kuntze）Exell. 在西非传统上用于伤口愈合。最近，已证明干叶的水提物可刺激人角质形成细胞和真皮成纤维细胞的增殖。在基于生物测定的分级分离中，鉴定出鞣花单宁根皮苷（1）、柯里拉京（2）、呋喃苷（3）、类黄酮槲皮素-3-O-β-D-葡萄糖苷（异槲皮苷）、山柰酚-3-O-β-D-葡萄糖苷（山柰苷）、槲皮素-3-O-D-芦丁苷（芦丁）、没食子酸、甲基没食子酸、咖啡酸、绿原酸、3,5-二咖啡酰奎宁酸和咖啡酰马来酸（phaselic 酸）首次在 P. muellerianus 中被鉴定出来。结果表明，根皮苷是水提物的主要成分。根据 ICH 指南（ICH 兼容方案），已开发并验证了适合 P. muellerianus 提取物（甲醇/水，70/30）中根皮苷质量控制的合适分析方法。根皮苷和呋喃苷增加了人皮肤细胞（真皮成纤维细胞 NHDF、HaCaT 角质形成细胞）的细胞能量状态，促使细胞增殖率更高，成纤维细胞比角质形成细胞更敏感。在 5 μM 时发现根皮苷对 NHDF 的刺激作用最大，在 50-100 μM 时对角质形成细胞的刺激作用最大。呋喃苷在约 50 μM 时刺激 NHDF，在约 150-200 μM 时刺激角质形成细胞。通过 LDH 释放测量到根皮苷的细胞坏死细胞毒性，在 20 μM 时对 NHDF，在 150 μM 时对角质形成细胞。呋喃苷的毒性（小于根皮苷）在>400 μM 时观察到。呋喃苷和根皮苷分别在 50 μM 和 5-10 μM 时刺激 NHDF 胶原的生物合成。在 105 μM 时，根皮苷显着刺激 NHEK 的分化，而呋喃苷对 involucrin 和角蛋白 K1 和 K10 的表达影响较小。该研究清楚地证明，来自 P. muellerianus 的亲水性提取物，特别是先导化合物根皮苷，对真皮成纤维细胞和角质形成细胞具有刺激作用，导致细胞增殖、屏障形成和细胞外基质蛋白的形成增加。从这些发现中可以看出，此类提取物的传统临床应用似乎是合理的，可用于伤口愈合。
