Panax Ginseng: Current Perspective Origin and Classification Panax ginseng (C. A. Meyer) is a Latin name given for ginseng root. The Panax, is the genus and ginseng is the species. The word of Panax comes from Greek word "Pan" meaning all and "akos" meaning cure, therefore in combination it means curing all (panaceas). Moreover, the words of ginseng comes from Chinese ideogram "Gin" meaning Man and "Seng" meaning Essence, therefore in combination it can literally be translated for " Crystallization of the earth in the form of a man (WWW1). P. ginseng belongs to Araliaceae family in which there are two genuses identified to have similar pharmacological effects both in animal and human clinical test. Panax includes both Panax ginseng CA Meyer, Panax quinquefolius (American ginseng), Panax japonicus CA Meyer, Panax notoginseng Burkill (Ngan et al., 1999) and Panax vietnamensis Ha et Grushv (Duc et al., 1996). The other genus is Eleutherococcus senticosus. The latter genus, however, has only a distant relationship with the previously mentioned genus because each genus consists of totally different compositional/chemical make-ups from each other. However, an abrupt reduction in the natural supply of ginseng as well as the difficulty in its cultivation led to the intense research on Eleutherococcus senticosus which resembles remedial efficacy of ginseng (Brekhman and Dardymov, 1969) The plant of P. ginseng is indigenous to Eastern Asia and is cultivated in China, Korea and Japan. The plant of P. quinquefolium is native to eastern and central North America and is cultivated nowadays in a modernized way throughout Wausau, Wisconsin (WWW3). Whereas the plants of Eleutherococcus senticosus is abundantly growing in the wild of Siberia and farmed as well in the Far East. Cultivation Review The plants of P. ginseng and P. quinquefolium are slow growing perennial herbs while the plant E. senticcosus is a woody/thorny plant. The seed of Panax is dormant for 18 month before germination can take place. In order to grow well, the plant requires a loose, rich soil with heavy mulch of leaves and about 80 per cent shade provided artificially. Moreover, ginseng plants should be sterilized and fertilized several times in a year. It is vulnerable to blight and insects. The plant has three leaflets in a seedling and five leaflets in a mature plant, with leaf number correlating with the age of the plant. At the first year of growth, the plant resembles a strawberry plant and develop flowers berries and seeds in the following years. It is suggested that at the age of 4 to 6 years ginseng root can then be harvested since at this time the root will contain optimal commercially valuable compounds (WWW4). Propagation by seed is the most common method although propagation by cuttings of the root has successfully been practiced. The root grows 3 - 5 times its starting weight each growing season (WWW7). Macroscopical Treatment/Processing after Harvest The treatments given to ginseng roots after harvest influence the visual characteristic of commercial ginseng. Somehow, tradition and the available knowledge of processing heavily influence the type of further processing of ginseng root. The root of Panax ginseng is usually steamed fresh after harvesting and then dried until the colour changes from white to Red and becomes known "Red ginseng". Further more, the root of Panax quinquefolius is usually just cleaned and dried after harvesting and retains the whitish colour of the root (WWW8). Root Composition Table 1 shows a typical composition of commercially dried ginseng. No Composition Percentage (%) 1 Water 10.0 2 Protein 12.2 3 Carbohydrate 70.0 4 Fiber 4.2 5 Fat 1.0 6 Ash 2.6 Source: Wang, 1996 Plant Cell Culture for Ginseng Production The demand for ginseng has increased dramatically worldwide and ginseng becomes very expensive because of its long-term conventional (5-7 years) and troublesome production cycles. The annual turnover of ginseng in the United States was $98 million with a growth rate of 26% (Vogler et al., 1999). Therefore, plant cell and tissue culture methods have been explored as potentially more efficient alternatives for the mass production of ginseng and its active components. Research into ginseng cell and tissue culture started in the early 1960s and commercial application has underway since the late 1980s. The powder and extracts from ginseng cell culture were used to make health foods, drinks and cosmetics. The ginseng culture has continued to attract considerable research and development effort in recent years and scientists seek to understand and optimize the culture conditions (Wu and Zhong, 1998). Earlier Furuya et al. (1984) reported that P. ginseng callus produces almost the same pharmacologically active saponins, ginsenosides as that of cultivated ginseng root. In a 30-liter jar fermentor culture, the increase of the growth ratio and dry weight were not accompanied by an increase of the saponin content. Using MS medium minus NH4NO3 and plus 0.5% glucose and 2% sucrose and 2% sucrose added after 2 weeks of culture resulted in a higher growth ratio and higher dry weight than using regular MS medium containing 3% sucrose. Effects of application sole nitrate (NO3-) and in combination with ammonium (NH4+) on production of ginseng saponin and polysaccharides by suspension cultures of panax ginseng were observed by Liu and Zhong (1997). The results indicated that the specific production (content) of ginseng polysaccharide was not significantly affected by alteration of the N source and the saponin production was relatively higher within the initial N concentration of 5 mM with nitrate alone or a (NO3-)/(NH4+) ratio of 2:1. Subsequently, the effects of inorganic phosphate on cell growth, accumulation ginseng saponin and polysaccharide together with nutrient utilization were also observed by Liu and Zhong (1998). The crucial concentration of phosphate for cell growth of Panax ginseng and Panax quinquefolium was 1.04 and 0.65 mM, respectively. Zhong and Wang (1996) investigated effects of Cu2+ in the medium on improvement of cell growth and production of ginseng saponins in suspension cultures of Panax notoginseng. It was found that with an increase of initial medium Cu2+ concentration within the range of 0 - 6.0 mM, the cell growth was greatly improved. The highest productivity and yield of ginseng saponins and polysacharide were obtained at 6.0 mM of initial medium Cu2+. Wu and Zhong (1998) reviewed all the conditions necessary for conducting cell culture in a bio-reactor including cell line induction and selection, reactor design operation, medium optimization and production process strategy and agreed that more stable quality of active ingredients would be obtained. The authors also noted that saponin content three times higher than in the callus of normal roots can be obtained from induced cell lines of crown galls of ginseng root infected with Agrobacterium tumefaciens. Further more, selection of cell line of P. ginseng that is resistant to chephazolin (antibiotic) resulted in the isolation of cell line Ic (chep) that produced 2.3 times greater amounts of ginsenosides than its non selected counterparts. The selected cultures did not decrease their enhanced production of ginsenosides for three years (Bulgakov, 1996). Recently in the era of panax biotechnology, Kevers et al. (2000) have proven the possibility of programming somatic embryogenesis of P. ginseng in liquid cultures. Further, the author confirmed that production of somatic embryos could be regulated by the manipulation of polyamine levels and metabolism either by using exogenous polyamines or their specific metabolic inhibitors. Identification, Quantification and Isolation Ginseng is not included in the Generally Recognized as Safe (GRAS) list nor has the US government set guidelines for manufacture and quality control of commercial ginseng preparation and hence there is no a single fixed method to analyze the ginseng composition. Although, it seems compulsory for ginseng to contain at least 1.5 % (w/w, as ginsenoside, Rg1) (Bluementhal et al., 1998). Nevertheless, Panax ginseng is mentioned in the pharmacopoeial monographs of the following countries as such Australia, China, Chech Republic, France, Japan, Russia, German and Switcherland (Newall at al, 1996). Separation, purification and identification of saponins are not so facile task. The active principle of ginseng saponin has never been confirmed chemically and pharmacologically until recently. Numerous ginsenosides were reported as hydrolysed saponins from Ginseng root. Genuine aglycones and the main common sugar component of the ginsenosides were 20S-protopanaxadiol, 20S-protopanaxatriol, oleanolic acid and glucose. Chemically it is characteristic that almost all the members of ginsenoside Rx except Ro are dammarane type triterpenoid saponin. Ginsenoside is an oleanane type triterpenoid saponin (Samuelsson, 1999). Earlier Hiai et al. (1975) developed a colorimetric method to indentify ginseng saponin from ginseng extract that would give red-purple colour when reacted with a mixture of vanillin and sulfuric acid and the quantity was determined accordingly by spectrophotometer. However, this method had difficulties in obtaining good recovery when isolation and recrystallization. Moreover, Otsuka et al.(1977) used Droplet Counter Current Chromatography (DCCC) to obtain 220 fractions from 25 mg of saponin samples eluted with solvent system: CHCl3: MeOH: n-PrOh: H2O (volume ratio: 45: 65:6:40). The absorbance of ginsenoside Rb,c,d,e,f and g could be clearly plotted and quantified by using a standard chart. The quantity of those ginsenosides varied from one variety to the other. Further, Liberti and Marderosian (1978) combined spectrodensitometer coupled with density computer with TLC methods in identifying ginseng composition. The spectrophotometric was for total saponin determination while the TLC was for individual ginsenoside and total saponin. The quantity of each composition was determined by Standard Chart's developed by evaluating area under the curve plotted accordingly to Beer's law. Plates were developed in methanol-chloroform-1-buthanol, dried, sprayed with mixture of p-anisaldehyde and concentrated sulfuric acid, heated and scanned at 600 nm. Transfer of samples into butanol solution avoided detection of free sugars. The recovery rate from this method was above 95% and in terms if total saponin, both the spectrodensitometry and TLC were in agreement. Nevertheless, there was variation in ginsenoside content of commercial ginseng. When the ginseng powder was reduced to 40 - 60 meshes in granulation, the evaluation became less confirmatory. This suggested that ginsenosides were sensitive to the handling and mechanical abuses during manufacturing. Two dimensional thin layer chromatography and spectrophotometry (Lui and Staba, 1980) could be used to do ginseng determination. This method was capable of measuring the weight and molar ration of each ginsenoside and separation of (20S)-protopanaxatriols (Re, Rf , Rg1 and Rg2) were clearly resolved. High-performance thin layer chromatography (HPTLC) was developed recently by Vanhaelen-Fastre et al. (2000). In this method, thionyl chloride was applied as the chromogenic agent. It appeared to suppress the ginsenosides diffusion into the reagent solution and/or irregular spreading and to increase reproducibility. The resolution between ginsenosides Rd and Re was markedly increased on this method. Additionally, this method offers simplicity of extraction and the low detection and quantification limits without a derivatization step. Considering that most of TLC methods are unable to separate Rb1/Rb2, Re/Rf and Rg1/Rg2, Soldati and Sticher (1980) introduced an HPLC method to separate ginsenosides from Panax ginseng. The samples were extracted with Extrelut where the aqueous samples were brought into a column filled with granulated carrier material and spread as a stationary phase on the porous matrix, then the column was eluted with n-butanol saturated with water. After evaporation of n-butanol, the dried eluted samples were injected into a reverse phased HPLC having (Bondapak C18. The eluent for Rb1, Rb2, Rc, Rd, Rf, and Rg2 were acetonitrile-water (30:70, 2 ml/minute) and for Re and Rg1 (18:82, 4 ml/ minute). Five types of ginsenosides were separated with Rd having the longest retention time (1614 seconds) and their quantities were determined by standard curve calculated based on the peak or area. Apart from long running time, HPLC method lacked of sensitivity due to the low UV Maximum (203 nm) at which lots of compounds, solvents, oxygen, etc absorbed UV light and they could lead to drifting of base line (Corthout et al., 1999). Methods of identification of ginsenosides from panax extracts will continue to evolve with time. One of the recent method is combining High Performance Liquid Chromatography and Mass Spectrometry (HPLC/MS). The advantage of this method is its capability to separate intact underivatized ginsenosides effectively from ginseng extract due to the fact that the column contained materials ranging from reversed phase silica and ion exchange packaging to specialized carbohydrate-containing aminopropyl functional groups (Chan et al., 2000). The method could also determine the source of ginseng source based on the measurement of ginsenosides, Rf and 24R-pseudoginsenoside F11. The HPLC/MS approach enormously increased detection sensitivity of by over 1000 times. Above-mentioned methods rely on acid hydrolysis of ginseng extracts. Acid hydrolysis gives limited yields and causes epimirization, cyclization and hydration of 20S-panaxadiold and 20S-panaxatriol. Therefore, additional oxygen/alkaline cleavage has been developed to counteract this problems (Cui, 1995). Being oxygen dependent, products of alkaline cleavage can thus be separated and identified. Quantification can be performed on Gas Chromatography. Further, in order to combat the illegal practice of disguising or adulterating Panax ginseng or other adulterants as Panax quinqufolius, Ngan et al. (1999) developed DNA finger printing profiles characteristic of P. ginseng and P. quinqofolius from AP-PCR and RAPD. This eliminated the large sample size for chemical analysis and limitation of chemical analysis in growth conditions and post-harvest treatment. The finding revealed that ITS1-5.8S-ITS2 was consistent among panax species and hence the restriction fragment length polymorphism (RFLP) of this region could be used to authenticate and to differentiate panax species from each other. There was a need to isolate particular ginsenoside, especially Rb1 to investigate its single pharmacological effect. Previous paragraph showed that ginseng root contains a number of dammarane types of ginsenoside together with oleanane saponins, the isolation of individual saponins is quite tedious and requires repeated column chromatography or preparative HPLC. Fukuda et al. (1999) established a simple reproducible purification method for ginsenoside Rb1 using an immunoaffinity column conjugated with anti-ginsenoside Rb1 monoclonal antibody. ElISA procedures and Western Blotting were applied in this method. The antibody was developed from mouse IgG. This latter method could also be used to isolate and separate any ginseng extract for other ginsenoside types and to survey low concentration of ginsenoside Rb1 of plant origin and/or in experimental animals and humans. Before leaving this identification section, it would be worthy to briefly mention that when animals or humans are subjected to clinical experiments it is possible to identify, isolate and measure the unused ginsenoside in biological fluids. Several studies have been reported in this matter based on labeling ginsenosides. Previous methods developed were either low in sensitivity, interfered with by cholesterol or other compounds contained in biological samples or were limited in use because of scarcity of radio active ginsenosides. Chen and Staba (1980) used gas liquid chromatography to assay individual ginsenoside and sapogenins in rabbit plasma and urine samples. A flavonoid, panasenoside, and a sterol, stigmasterol were used as internal standards for ginsenosides and their sapogenins, respectively. Measuring the linear relationship between peak and height ratio, the sensitivity of 0.2(g and 0.1(g were obtained for ginsenosides and sapogenins, respectively. A schematic diagram for ginseng extraction is shown in Figure 1. The isolation and elucidation of ginseng saponins in the figure was conducted through repeated chromatography with appropriate solvents. Biosynthesis and Structures of Ginsenosides TLC, GLC, and InfraRed instruments were used in isolation and elucidation of ginsenosides' structures (Sanada et al., 1974). Ginsenoside Ro Structure of Ginsenoside Rb1 Ginsenosides Rb1 resembles to 20S-protopanaxadiol-3-[O-(-D-glucopyranosyl(1-2)-(-D-glucopyranoside]-20-[O-(-L-arabinopyrranosyl(1®6)-(-D-glucopyranoside]. Structure of Ginsenoside Rb2 Ginsenoside Rb2 is built as 20S-protopanaxadiol-3-[O-(-D-glucopyranosyl(1-2)-(-D-glucopyranoside]-20-[O-(-L-arabinopyrranosyl(1®6)-(-D-glucopyranoside]. Structure of Ginsenoside Rc Ginsenoside Rc is built from 20S-protopanaxadiol-3-[O-(-D-glucopyranosyl(1-2)-(-D-glucopyranoside]-20-[O-(-L-arabinofuranosyl(1®6)-(-D-glucopyranoside]. Structure of Ginsenoside Rd Ginsenoside Rd is built from 20S-protopanaxadiol-3-[O-(-D-glucopyranosyl(1-2)-(-D-glucopyranoside]-20-[O-(-D-glucopyranoside]. Structure of Ginsenoside Re Ginsenoside Re is built from 20S-protopanaxatriol-6-[O-(-L-rhamnopyranosyl(1®2)-(-D-glucopyranoside]-20-O-(-D-glucopyranoside. Structure of Ginsenoside Rf Ginsenoside Rf is built from 20S-protopanaxatriol-6-[O-(-D-glucopyranosyl(1®2)-(-D-glucopyranoside. Structure of Ginsenosides Rg1 The structure of this saponin has been established as 6,20-di-O-(-glucosyl-20S-protopanaxatriol. Hydrolysis of Ginsenosides Rb1, Rb2 and Rc with concentrated HCl at room temperatures and followed by dehydrochlorination lead to 20R- and 20S-protopanaxadiols if which undergo Smith's degradation will afford genuine sapogenin of Rg1 (Nagai, et al., 1971). Structure of Ginsenosides Rg2 In order to obtain this type of ginsenoside, Kaku and Kawashima (1980) developed sequential extraction with dichloromethane, methanol, precipitation and evaporation and recrystalisation. The structure of ginsenoside Rg2 was established as follows: 20S-protopanaxatriol-6-O-(-L-rhamnopyranosyl(1®2)-(-D-glucopyranoside. Structure of Ginsenoside Rh Ginsendoside Rh can only be obtained from minor saponins of red ginseng by mild hydrolysis of ginsenoside Rb, Rc, and Rh and it does not exist in white ginseng. Due to similarity in structure, Atopkina et al. (1997) attempted the synthesis of ginsenoside from Betulafolienetriol [dammar-24-ene-3-(,12(, 20(S)-triol] isolated from birch leaves. In five steps, condensation of the 12-O-acetylderivative of 20(S)-protopanaxadiol [dammar-24-ene-3-(,12(, 20(S)-triol] with tetra-O-acetyl-(-D-glucopyranosyl bromide in the presence of silver oxide in dichloroethane, followed by deprotection with sodium methoxide in methanol results in the formation of the 3-O-(-Dglucopyranosyldammar-24-ene-3-(, 12(, 20(S)-triol which is identical with natural ginsenoside Rh. Other Ginsenosides To date about thirty glycosides have been isolated from roots of Panax ginseng and interest has been focused on testing biological activity of individual ginsenoside rather than in combination as in Chinese medicine. Anufriev et al. (1997) successfully synthesized Rg3 by glycosylation of 12(-acetoxy-dammar-24-ene-3(,20(S) diol with hepta-O-acetyl-(-sophorosyl bromide under catalysis by Ag2CO3. The pure form of Rg3 was obtained after deprotection and chromatographic purification. The compound was fully characterized by 1H and 13C NMR spectroscopy. Earliear Sanada and Shoji (1978) also reported the isolation of Ginsenoside Rb3 as 20S-protopanaxadiol-3-[O-(-D-glucopyranosyl(1-2)-(-D-glucopyranoside]-20-[O-(-L-xylopyrranosyl(1®6)-(-D-glucopyranoside]. This only differs from ginsenoside Rb2 in the sugar moiety attached on the carbon 20 of the aglycone. Chemical and Biological Activities of Ginseng Extracts and their individual component Being one of the most popular herbal medicines, a number of health claims or activities have been made public about ginseng extracts and its constituents. Ginseng extract can exert such widely different pharmacological actions because it contains more than 200 different compounds which exert different activities (Soldati, 2000). Traditionally, ginseng use has been divided into two categories: short term, to improve stamina, concentration, healing process, stress resistance, vigilance and work efficiency in healthy individuals and long term, to improve well being in debilitated and degenerative conditions especially those associated with age. This paper from now on will review and evaluate the evidence of such claims scientifically. Many Chinese believe that life-prolongation effect of ginseng during Liang Dynasty in China may be due to the preventive activity of ginseng against the development of cancer. Yan and Choi (1998) conducted consumer survey for three years at 4634 people over 40 years of age to support this hypothesis. The subjects were asked to questions regarding age at ginseng initial intake, frequency, ginseng type and duration of intake. A multiple logistic regression was used to estimate relative risk when controlling simultaneously for covariates. The finding was that ginseng consumer had decreased risk for cancer compared with the non-consumer. Frequency of ginseng consumption and dosages also decreased the risk. Ginseng Effect as Anti-Oxidant and Oxygen Link A research was conducted by Kitts et al. (2000) to determine the antioxidant properties of ginseng extract from Panax quinquefolius. It was found that ginseng extracts exhibited effective antioxidant activity in both lipid and aqueous media by both chelation of metal ions and scavenging of free radicals such as DPPH, Peroxide and Hydroxide. Ginseng extracts also inhibited the non-site specific DNA strand breakage cause by Fenton agents and suppressed the Fenton induced oxidation of a 66 Kd soluble protein obtained from mouse brain over a concentration of 2 - 40 mg/mL. Moreover, Chang et al (1999) investigated effects of ginseng saponins on the induction of catalase gene expression and superoxide dismutase (SOD) that converts superoxide radicals to H2O2 that in turn is broken down to water and oxygen by catalase. It was found that panaxadiol (ginsenoside Rb2) increased the transcription of SOD genes about 2-3 fold. Hence, cell viability could be maintained by lowering the level of oxygen radical generated from intracellular metabolism. Ginseng reportedly (Gillis, 1997) increased resting oxygen uptake and oxygen transport in elderly subjects and significantly increased the capacity for mental arithmetic and logical deduction in healthy normal subjects. On a double blind cross over study, it was found that at a fixed workload ginseng decreased oxygen consumption, carbon dioxide production and plasma lactate. Brekhman and Dardymov (1969) termed these non-specific tonic effects of ginseng as adaptogenic. Intravenouse injection of total ginsenosides into dogs rapidly decreased the peak value of left ventricular pressure and aterial systolic pressure. The heart rate and renal arterial blood flow were also decreased, whereas renal vasoresistance was markedly increased by treatment with total ginsenosides (Tang and Eisenbrand, 1992). Ginsenosides also increased the myocardial oxygen tolerance of animals to hypoxia. This was associated with a decrease in myocardial oxygen consumption during hypoxia. Lower heart rate and quicker return to normal was observed when standardised ginsenosides were applied to participating atheletes (Dharmananda, 1997) due to improved oxygen utilization and faster clearance of lactate. Effects of Ginseng on Nitric Oxide Link In order to find out the antiasthmatic effect of P. ginseng, Tamaoki et al. (2000) studied relaxant responses of human bronchial strips under isometric condition in vitro and directly measured the release of nitric oxide (NO) by amperometric sensor. The addition of ginsenoside (210(g/mL) relaxed the tissues precontracted with acetylcholine for about 67%. The addition of ginsenoside to the medium containing bronchial tissue dose-dependently increased NO-selective electrical current and this effect was greatly attenuated by the epithelial removal or CA2+ free medium. Both Gillis (1997) and Nocerino et al (2000) reported that ginseng had clinically positive effect on sexual impotence and aphrodisiac properties. Nitric oxide (NO) was released from nonadrenergic non-cholinergic (NANC) nerves and relaxed both human and rabbit penile corpus thus permitting penile erection. Ginsenosides enhanced formation of cirtulline from added arginine, implying enhanced synthesis of NO. Effects of Ginsenoside on Cancer and Tumor Based on the information that ginsenoside Rh2 of Panax ginseng inhibited the proliferation of cancer cells and induced their reverse transformation in cultures of Morris hepatoma cells, Xiaoguang et al. (1998) conducted an experiment on Wistar rats transplanted with sarcome S180 and B16 melanoma. The growth of transplantable sarcoma S180 and melanoma B16 and tumor cell proliferation were inhibited due to the inhibition of DNA, RNA and protein synthesis. Further the group also found that the ginsenosides might enhance body cell immune function by promoting the transformation of T lymphocyte and inhibited rat liver microsome lipid peroxidation significantly in a dose dependent manner. Brief exposure of rat cultures to excess glutamate caused extensive neural death. Nevertheless, glutamate induced neuronal cell damage was reduced significantly by pretreatment with ginsenosides Rb1 and Rg3 as it was investigated by Kim et al. (1998). Ginsenosides Rb1 and Rg3 inhibited the overproduction of nitric oxide, which routinely followed glutamate neurotoxicity and preserved the level of superoxide dismutase (Chang et al., 1999) in glutamate treated cells. Furthermore in rat cultures treated with glutamate, these ginsenosides inhibited the formation of malondialdehyde, a compound that is produced during lipid peroxidation and diminished influx of calcium. It was concluded that ginsenosides Rb1 and Rg3 exerted significant neuroprotective effects on culture cortical cells. The protective action of ginsenoside Rb1 on acute nephrosis induced by puromycin aminonucleoside (PA) was observed Lim et al. (1998). Ginsenoside Rb1 was able to suppress the formation of phosphatidylcholine hydroperoxide in the plasma, liver and kidney along to increase glutathione peroxide activity in the blood. Further, Tang and Eisenbrand (1992) reported that total saponin from P. ginseng prevented the stem cell killing effects of harringtonine in treatment of L1210 leukemia in mice. Thus, the use of ginseng as adjuvant in tumor chemotherapy would be much more relevant. The water extract of ginseng inhibited the side effects of anticancer agents 5-fluorouracil and mitomycin C in decreasing the number of leukocytes, urine flow, renal plasma flow, glomerular, filtration rate and urinary excretion of sodium. However, cytosin arabinoside induced damages to the hematopoietic precursor cells in mice were enhanced by the intraperitoneal or oral pretreatment of total saponins. Effects of Ginsenoside on Central Nervous System and Hormone Brekhman and Dardymov (1969) earlier carried out an extensive study on the stimulant effect of ginseng extract towards fatigues and stress. For the test, male mice were either placed on a continuously moving endless rope passing a descending manner through a closed vertical box and fixed on their back for 24 hours at 450C or hung on their side for 16 - 26 hours, respectively. The results indicated that ginsenoside Rd (0.151 mg) had major influence in prolongation of the time to physical collapse through fatigue and 2.5 mg of mixed ginsenosides inhibited a major change in weight of the adrenals, thymus, spleen and the thyroid demonstrated in control animals. More experimental studies (Nocerino et al., 2000) confirmed that ginsenoside saponins of ginseng extract enhanced resistance to X-irradiation, viral tumor load, temperature stress, hyperbaric hyperoxia, physical exercise, augment work capacity and increased swimming time in rats. Many of these activities have been attributed to a corticosteroid-like action of ginseng. The author also noted that oral administration of ginseng Rb1 at a dose of 20 mg/kg for 3 days improved learning and memory in rats performing a maze task and shuttle box active avoidance test. The efficacy of Rb1 in enhancing short-term learning and memory included a direct stimulation of synapstosomal uptake of the precursor for Acetylcholine (ACh) synthesis and the release of ACh via a novel yet unknown mechanism (Wang, 1996). The ACh would be able to transmits impulses from nerve fibres (Cernetig, 1993). While, Yuan et al. (1999) noted that Rb1, Rg1 and Re prevented scopolamine induced memory deficits. Further Gillis (1997) reported that ginsenoside Rg2 could particularly reduce ACh-evoked release of catecholamines from bovine adrenal chromaffin cells. In this way, ginsenosides might reduce elevated circulating catecholamine concentrations associated with various forms of stress in humans. In a double blind placebo controlled study conducted by Sotaniemi et al (1995), 36 newly diagnosed noninsulin dependent diabetes mellitus (NIDDM) patients were treated for 8 weeks with ginseng (100 or 200 mg) or placebo. Glucose balance, serum lipids, aminoterminal propeptide (PIIINP) concentration and body weight of the patients were measured along with psychophysical test. The result indicated that the 200-mg dose of ginseng improved glycated hemoglobin, serum PIIINP and physical activity. Other ginseng group showed mood elevation, improvement in psychophysical performance and reduction in fasting blood glucose (FBG) and body weight. While the placebo group showed reduction in body weight and alteration serum PIIINP. When ginseng extract and deer antler extract were applied to experimented mice, luteinizing hormone secretion increased about ten fold (Dharmananda, 1997). This hormone influences the menstrual cycles of in females and it stimulates testosterone secretion. Luteinizing hormone is produced by the pituitary gland so it was proposed that mixture of ginseng and deer antler extracts influenced the hypothalamus and pituitary and adrenal axis. A time course study, on the effect of ginsenoside Rb2 on lipid metabolism in rats showed that the first phenomenon to be observed was a stimulation of glucose-6-phosphate dehydrogenase activity beginning 4 hour after the adminstration. An increase in in the lipolytic activity of lipoprotein lipase was observed 4 hour after intraperitoneal administartion of ginsenoside Rb2 and reached maximum 16 hour after treatment. Similar effects of the ginsenoside in lowering the serum total cholesterol level and serum low-density protein cholesterol level were also observed in white leghorn females. In addition the activities of (-hydroxy-(methylglutary-CoA reductase and cholesterol-7(-hydroxylase in liver were inhibited (Tang and Eisenvrand, 1992). Effects of ginsenosides on Transport/Absorption Mechanisms Ginsenosides are amphiphilic in nature and have the ability to intercalate into the plasma membrane. This leads to changes in membrane fluidity and thus affects membrane function and eliciting a cellular response. There is evidence to suggest that ginsenosides interact directly with specific membrane protein. The effects of ginsenosides on membrane channels show similarities to steroid hormones that modulate rapid Ca2+ influx in several tissues although ginsenoside Rc might inhibit Ca2+ influx through voltage gated Ca2+ channels in adrenal chromaffin cells (Yuan et al., 1999). Alcoholism is a serious medical, social and economic problem facing almost all-human societies worldwide. Medical interventions in the field of alcoholism are primarily aimed at relieving the consequences of alcohol withdrawal syndrome, arresting alcohol drinking and maintaining sobriety for as long as possible. Pharmaco-therapy is conceived to provide a substantial contribution to these goals, facilitating the psychological support and social rehabilitation of alcoholic patients. Recent experimental evidence and critical re-examination of empirical data from traditional medicines suggest that novel pharmacological approaches for treatment of alcoholism and alcohol abuse may stem from natural substances. Ginseng was proposed to accelerate alcohol metabolism and lower blood alcohol levels by increasing ADH and alter alcohol adsorption from the gastrointestinal tract of rats (Carai, et al., 2000). At different study, Yamasaki (1996) investigated that using sheep erythrocyte ginsenonsides Rc, Rb1, Rb2, Rf, Rg1 and Rg2 had potent stimulatory activity against 2-deoxy-glucose (2-GD) uptake. The stimulatory effect was significantly reached at 100 (g/ml of ginseng extract. The hypoglycemic action of ginsenosides were reported (Vacek and Sun, 2000) to lower glucose levels in diabetic patients. It was presumed that the beneficial effect of ginseng in reducing hyperglycemia in diabetics might result from a stimulatory effect of ginsenosides on residual islets. Effects of Ginsenosides on Immunity Ginsenoside Rg1 was shown to increase both humoral and cell mediated immune responses. Spleen cells recovered from ginsenoside treated mice injected with sheep red cells as antigen showed significantly higher plaque forming response and hemagglutinating antibody titers. In addition, Rg1 increased the number of antigen reactive T-helper cells, T-lymphocytes and NK cells (Yuan et al., 1999). Gastric infusion of ginsenoside into mice caused stimulation of the phagocytic function of the reticuloendothelial system, an increase in serum-specific antibodies and IgG contents and also an increase in the relative percentages of B cells. Given orally, intraperitoneally or subcutaneously ginsenosides were effective against immunodeficiency induced by cyclophophamide in mice (Tang and Eisenbrand, 1992). Tips for Ginseng Consumption It is recommended that a short break of one to two weeks when a person is using ginseng. Traditionally, dossage recommendation differs between the short-term use in healthy individuals and the long-term use in elderly or debilitated person (Newall, et al.,). Short term (For the young and healthy), 0.5 - 1.0 g root daily as two divided doses for a course generally lasting 15 - 20 days and with a root free period of approximately two weeks between consecutive courses. Doses are recommended to be taken in the morning, two hours before a meal and in the evening, not less than two hours after a meal. Long term (For the old and sick), 0.4 - 0.8 g root daily can be taken continuously. Side Effects of Ginseng Interaction Many medicinal herbs and pharmaceuticals are therapeutic at one side and toxic at another. Doses of 15 gram of Panax ginseng root material daily were reported to associate with depersonalization and confusion, while depression was reported after consuming more than 15 gram daily. The LD50 of ginseng root in mice was reported to 10-30 g/kg (Gillis, 1997). Ginsenoside Rb1 was the most toxic amongs ginsenoside after intrapertinoneal administration to mice (Tang and Eisenbrand, 1992). Patients having ginseng abuse syndrome experienced hypertension, nervousness, sleeplessness, skin eruptions, morning diarrhea, euphoric and agitated (Vogler et al., 1999). Interaction between herbs and drugs may increase or decrease the pharmacological or toxicological effects of either component. Shader and Greenblatt (1985) reported that patients had developed headache and tremor as a result of interaction between ginseng and Phenelzine. Moreover, Jones and Runikis (1987) noticed that patients who previously ingested bee pollen and had unipolar depression were more likely to develop mania as the result of such drugs and herbs interaction. Further, Lee et al. stated that interaction of ginseng and alcohol caused increased alcohol clearances. Later it was noted as well that in mice ginseng increased the activity of alcohol dehydrogenase and aldehyde dehydrogenase. A case of decreased International Normalised Ratio (INR) was also reported by Janetzky and Morreale (1997) on 47 year old man who was on warfarin medication and took Gisana( three times daily to improve his energy level. When freed from Ginsana the INR level was back t o normal. Pharmacokinetic studies Tang and Eisenbrand described the pharmacokinetic study of ginseng in detail based on the ginsenoside Rg1 and Rb1. Ginsenoside Rg1 was absorbed rapidly from the upper parts of digestive tract in rats and it was not found in brain. Excretion into urine and bile occurred at a ratio of 2:5. It was not metabolized to a significant extent in the liver but decomposition and metabolism in rat stomach and large intestine were comfirmed. In a stimulated gastric acid medium, ginsenoside Rg1 was degraded to some prosapogenin. On the other hand, only little ginsenoside Rb1 was absorbed from digestive tract after oral administration of 100 mg/kg to rats. Ginsenoside Rb1 was gradually excreted into urine but not bile. In digestive tract, unadsorbed ginsenoside Rb1 was rapidly decomposed and metabolised in tlarge intestine and also degraded into prosapogenin in stimulated gastric acid. Efficacy of Ginseng Vogler et al (1999) conducted systematic literature searches to identify all randomized controlled trials (RCTs) in determination of true efficacy of ginseng. Total of 57 RCTs on ginseng were retrieved and 16 trials met the criteria. The finding revealed that ginseng may have beneficial effects on psychomotor performance and cognitive behavior but had contradictory evidence for ginseng to improve physical performance and immunological parameters. Therefore, the widespread use of ginseng as an herbal remedy warrants more rigorous investigations to assess its efficacy and safety. Lots of researches about ginseng have been conducted all over the world. Nevertheless, the access to that researched information is limited due to language barrier and paper distribution amongst research institution. No doubts than several researches about ginseng overlapped or contradicted each other. This results in unresolved issue about ginseng and creates more confusion about its confirmed biological activities. Some researches are discarded from clinical consideration because of lacking controlled double-blinded studies. The other missed out the consistency by not providing clear information about ginseng materials used and hence the reproducibility of such result is doubtfully possible. Therefore, in the future standardization of ginseng material is the prerequisite for a constant pharmacological answer about ginseng. Conclusion In Vitro and in vivo pharmacology studies about ginseng in the past proved that ginseng contains biologically active compounds that have positive actions in a quites specific way at specific sites of human or animal subjects. Nevertheless, above all ginseng has been known for its long-term adaptogenic capacity and hence its usage as herbal medicine will be constant or continure at the rate of similar to other alternative medicines. Therefore, ginseng will be still a potential comodities in the future. Applying extensive chromatography methods developed for both extraction and identification of ginsenosides now can do standardisation of ginsenosides contents of ginseng. 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