Phytochemistry Reviews (2005) 4: 197–219 Ó Springer 2005 DOI 10.1007/s11101-005-3754-4 Chemistry and pharmacology of Gynostemma pentaphyllum Valentina Razmovski-Naumovski , Tom Hsun-Wei Huang , Van Hoan Tran, George Qian Li, Colin C. Duke & Basil D. Roufogalis* Faculty of Pharmacy, Herbal Medicines Research and Education Centre, A15, University of Sydney, N.S.W., 2006, Australia; *Author for correspondence (Tel.: +61-2-9351-2831; Fax: +61-2-9351-4447; E-mail: [email protected]) Key words: dammarane, gynosaponin, gynoside, Gynostemma pentaphyllum, gypenoside, herbal medicine, saponin, sterol Abstract In traditional Chinese medicine, Gynostemma pentaphyllum (Thunb.) Makino is a herbal drug of extreme versatility and has been extensively researched in China. The dammarane saponins isolated from Gynostemma pentaphyllum, namely gypenosides or gynosaponins, are believed to be the active components responsible for its various biological activities and reported clinical effects. This review attempts to encompass the available literature on Gynostemma pentaphyllum, from its cultivation to the isolation of its chemical entities and a summary of its diverse pharmacological properties attributed to its gypenoside content. Other aspects such as toxicology and pharmacokinetics are also discussed. In vitro and in vivo evidence suggests that Gynostemma pentaphyllum may complement the popular herbal medicine, Panax ginseng, as it also contains a high ginsenoside content and exhibits similar biological activities. Abbreviations: ara – arabinose; G. pentaphyllum – Gynostemma pentaphyllum; glu – glucose; i.g. – intra- gastric; i.m. – intramuscular; i.p. – intraperitoneal; i.v. – intravenous; LD50 – mean lethal dosage; lyx – lyxose; rha – rhamnose; s.c. – subcutaneous; xyl – xylose Introduction species of Gynostemma mostly growing in south- western China (southern Shaan Xi and areas south This review paper attempts to summarise the of the Yangtze River). The pentaphyllum species is existing literature on Gynostemma (G.) penta- the most prevalent and is dispersed throughout In- phyllum (Thunb.) Makino, with some reference dia, Nepal, Bangladesh, Sri Lanka, Laos, Myan- made to the authors’ own investigations. mar, Korea and Japan (Blumert and Liu, 1999). G. G. pentaphyllum is a perennial creeping herb of pentaphyllum is distributed naturally in mountain the genus Gynostemma. The plant belongs to the forests, mountain valleys, wood, scrub, stream Cucurbitaceae family which includes cucumber, banks, roadsides, bushes, shaded and humid places gourd and melons (Blumert and Liu, 1999). Com- at an elevation of 300–3200 m (China Pharmaceu- mon names of G. pentaphyllum include Jiaogulan, tical University, 1996). Qi Ye Dan, Gong Luo Guo Di, Pian Di Sheng Gen, The use of G. pentaphyllum has a history of Xiao Ku Yao, Amachazuru, Dungkulcha, Baan Ja about 500 years, with most of the reviewed mate- Kahn and Penta (Guo and Wang, 1993; China rial published in China. The book ‘Herbs for Pharmaceutical University, 1996). There are 21 Famine’ published in the Ming Dynasty (1368– 1644 A.D.) describes the herb’s usage as a vegeta- These authors contributed equally. ble, suitable as a food, or a dietary supplement 198 during famine rather than as a medicinal herb seasons are April to December (China Pharma- (Cheng, 1990). The renowned herbalist Li Shi-Zhen ceutical University, 1996; Blumert and Liu, 1999). (1578 A.D) later described G. pentaphyllum for The methods and optimal conditions for grow- treatment of haematuria, oedema and pain of the ing G. pentaphyllum are outlined in China Phar- pharynx, heat and oedema of the neck, tumours maceutical University (1996), Wang et al. (1996) and trauma (Wu, 1957; Li, 1985). According to and Guo and Wang (1993). A soil (pH in the range traditional Chinese medicine (TCM) principles, the 5.5–8.0, with an optimal pH of 6.5–7.0) more than taste and nature of G. pentaphyllum is slightly bit- 30 cm deep, rich in humus, nitrogen, phosphorus ter, neutral, warm, enhancing ‘Yin’ and supporting and moisture, is ideal for growing G. pentaphyllum. ‘Yang’ and suggested that the plant ‘would be used The optimal soil is well aerated and retains water, to increase the resistance to infection and for anti- such as a fertile sandy loam. G. pentaphyllum prefers inflammation’ (Blumert and Liu, 1999). Indica- to grow in a warm and humid environment. The tions include hyperlipidaemia, palpitation and viable air temperature is )15–41.5 °C. The optimal shortness of breath, chest congestion, tingling air temperature is 15–30 °C. The light factor is sensation in the limbs, dizziness, headache, for- important for growing G. pentaphyllum (Huang getfulness, tinnitus, spontaneous perspiration, et al., 2004). The plant prefers shady areas, with a general weakness, swelling of abdomen, Qi defi- viable relative illumination of 40–80% and optimal ciency of heart and spleen and stagnation of relative illumination of 65–75%. Under these con- phlegm and blood (China Pharmaceutical Univer- ditions, the output is the highest, the blossoming sity, 1996). Therefore, G. pentaphyllum has since and bearing of fruit is increased and the total been included in the most recent dictionary of saponin content is the highest (Guo and Wang, Chinese Materia Medica, where the plant is 1993; Wang et al., 1996). Wind damage is common prescribed in TCM for heat clearing, detoxifica- for this vine plant and can be reduced by tion, antitussive, heart palpitation, fatigue syn- introducing wooden sticks so that the plant can drome, chronic bronchitis and expectorant for intertwine itself. An ideal chemical fertiliser is a relieving cough (Wu, 1998; Blumert and Liu, 1999). nitrogen–phosphate–potassium compound (2–3 In Japan, it is indicated as a diuretic, antipyretic, times) during the active growth period. Pests such as anti-inflammatory and tonic (Lin et al., 1993; Chen ants and snails can be controlled by various com- et al., 1996; Tanner et al., 1999). mercially available products. G. pentaphyllum may The appearance of G. pentaphyllum consists of be collected when the vine reaches 2–3 m in length. slender stems of thin, soft leaves arranged like In the subtropics and tropics, the herb is collected fingers on a hand, bearing 3–9 leaves (usually 5–7 4–5 times a year. With optimal growing conditions leaves). The leaflets are long and pliable, broadest and luxuriance, collection may be every 20–30 days. below the middle and tapering to a point like a In a high-yield plot, 4000–5000 kg of dried herb lance. They are rough to the touch on both sides, may be collected every hectare (Guo and Wang, are coloured a deep green on the upper side and a 1993). In comparison, the cultivation of Panax (P.) light green underneath. G. pentaphyllum has male ginseng (Araliaceae) is highly specialised, requiring and female flowers on separate plants (Wang et al., a maturation period of up to seven years to attain 2004). The conical, multiple male flowers are 10–15 optimal medicinal properties (Lee, 1992). (up to 30) cm. The 5-cleft corolla of petals is pale Razmovski-Naumovski et al. (2005a) documents green or white. The female flowers are similar but the cultivation of Gynostemma pentaphyllum out- are much smaller than the male. The 2–3 cavity side its native China in Sydney, Australia. Some ovary is globe-shaped. The three styles and stigma research has also been undertaken in cell culture of are short and cleft into two parts. The fruit con- G. pentaphyllum (Zhang et al., 1989, 1995; Chang sists of a smooth, globular, small berry about et al., 2005). 5–6 mm in diameter and black when ripe. The two ovo-heart shaped seeds inside (~4 mm in diameter) are coloured greyish brown or deep brown. The Isolation and identification methods apex of the seed is obtuse and the base is heart- shaped. The flowering seasons in the northern Gynostemma total saponins, namely gypenosides hemisphere are March to November; the fruiting (or gynosaponins) exist mainly as dammarane 199 type-triterpene glycosides (Figure 1). Due to their raphy (HPLC) conditions are usually restricted to structural similarity, gypenosides are subjected to the use of acetonitrile under UV (<203 nm) or a variety of isolation and identification methods. refractive index. Identification has been achieved G. pentaphyllum (usually dried material) can be with liquid chromatography–mass spectrometry extracted with a variety of solvents, including (LC-MS) using acetylated gypenosides and 1H- water, methanol and ethanol for the more polar and 13C-NMR results via chemical degradation to material; ethyl acetate, petroleum ether and chlo- aglycone and sugar moieties and comparison with roform for the less polar material, using an open compounds of closely related structures (Nagai et system or under reflux. The separating methods al., 1981). Characterisation of the aglycone has include 1:1 partition with water and butanol, silica also been achieved with alkaline cleavage of the gel chromatography, alumina (Al2O3), sephadex gypenosides and gas chromatography–MS (GC– (LH-20), ODS, XAD-2 and other styrene co- MS) with trimethylsilated aglycones (Cui et al., polymers (Nagai et al., 1981; Takemoto et al., 1998, 1999). Spectral methods such as 1H–1H two- 1981, 1982; Kuwahara et al., 1989; Zhao et al., dimensional 1H correlation spectroscopy (COSY), 1995; Wang and Li, 1997; Wang et al., 1997a). 13C distortionless enhancement by polarisation Thin layer chromatography (TLC) provides the transfer (DEPT), homonuclear Hartmann–Hahn first identification profile of G. pentaphyllum.