Ginseng Flowers Stimulate Progesterone Production from Bovine Luteal Cells

American Journal of Chinese Medicine,  Summer-Fall, 2000  by Leang-Shin Wu,  Shiow-Yunn Sheu,  Cheng-Chun Huang,  Chih-Hsien Chiu,  Jan-Chi Huang,  Jenn-Rong Yang,  Wai-Xiong Lian,  Chih-Hao Lai,  Yuh-Pan Chen,  Jen-Hsou Lin

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Abstract: Our previous report first showed evidence that polysaccharides isolated from ginseng leaves obtained from Jilin, China possess luteotropic activities. In this study, we made further investigations on the root and flowers of Korean ginseng by means of the same bioassay system described briefly as follows. Frozen-thawed bovine luteal cells (1 x [10.sup.5] cells/ml/well) in M199 were incubated in 24-well culture plates at 37 [degrees] C in a 5% [CO.sub.2] incubator. Ten [micro]l of tested drugs with 1, 10 and 100 [micro]g/ml were added into each well. After 4- and 24-hr incubation, the media were harvested and assayed for progesterone by an enzyme immunoassay. The production of progesterone from cells is the indicator for evaluating the action of tested drugs. Results showed that hot water extracts of ginseng flowers (GF-1) with 10 to 100 [micro]g/ml significantly increased progesterone production, whereas those from ginseng root (GR-1) could not. Crude polysaccharides (GF-2) isolated from GF-1 is the active component and the small molecules (mw [is less than] 10,000 dalton) are excluded, indicating that the ginseng root has no luteotropic activities, but the polysaccharides of ginseng flowers have.

The root of Panax ginseng has been used clinically in Chinese herbal medicine to treat many kinds of chronic diseases including hepatitis and nephritis, to enhance immune-system activities, to increase hypothalamo-pitiutary adrenocortical functions and sexual ability (Wang, 1991; Fu et al., 1981; Lin et al., 1995; 1998a, b). Such diversity of pharmacological activities was also approved in research works (Saito, 1974; Rim, 1979; Gao et al., 1989; Hu et al., 1995; Lin et al., 1995; 1998a; Huh et al., 1998; But, 1999). For this reason ginseng has been called the `king of tonic medicine' in the Orient and a panacea in the West. Brekhman and Dardymov (1969) even introduced a concept of adaptogen to describe it. Adaptogen means something that can keep human beings or animals in a state of nonspecifically increased resistance (SNIR), which not only maintains homeostasis, but overcomes stress. Many Chinese medicinal herbs categorized in the tonic drugs, e.g. ginseng, san-qi (P. notoginseng), tsu-wu-jia (Ancanthopana senticosus) etc. have such actions (Lin et al., 1998b).

In recent years, some scientists, especially Japanese groups, have further investigated the functions of ginseng leaves and found that the leaves also possess some properties, as do the roots, such as anti-ulcerative, immunity enhancing activities etc. (Gao et al., 1984; Sun et al., 1992, 1994; Yamada, 1994). This suggests ginseng leaves may replace roots in some aspects of clinical use. Even more recently, the ginseng flower is also being focused on to develop its use, especially on the nutraceutical sector. Some commercial products have already appeared on the market in Korea, Japan, China and Taiwan. Meanwhile, very few papers concerning research works and clinical use of ginseng flower have been published in the scientific media.

In the present study, the luteotropic activities of ginseng flower and root were examined to evaluate their bioactivities.

Materials and Methods

Materials

Roots and flowers of Panax ginseng C. A. Meyer were cultivated and harvested in Korea and their dried products bought in Taipei, Taiwan. The culture media were purchased from Pharmacia Co. Ltd. Ovine luteinizing hormone (NIDDK-oLH-1-3, Bethesda, MD) was kindly provided by the National Hormone and Pituitary Program, USA. The EIA kits for progesterone were made in our laboratory (Wu et al., 1997). The bovine luteal tissues were obtained from Heng-Chun Station, Taiwan Livestock Research Institute. Other chemical compounds such as bovine serum albumin, collagenase, desoxyribonuclease I etc. were obtained from Sigma Chemical Co.

Preparation of Crude Hot Water Extracts and Polysaccharides from Roots or Flowers of P. Ginseng

One hundred grams of dried roots or flowers of P. ginseng were decocted with water (1 L) until the volume was reduced by half. The residue was then reboiled further after adding water as above. The two extracts were combined and lyophilized to obtain the water-soluble extracts. The extracts were named GR-1 for root extracts and GF-1 for flower extracts, respectively. Ten grams of GF-1 was refluxed with 500 ml MeOH for 1 hr and then centrifuged to get a MeOH-insoluble precipitate. This precipitate was dissolved in water and dialyzed against running water for three days. After the non-dialyzable portion was centrifuged at 8,000 rpm for 30 min, the supernatant was lyophilized to obtain crude polysaccharides (GF-2, yield 1.2 g). The MeOH-soluble fraction was evaporated to dryness. After that, the residue was redissolved in water and lyophilized to obtain GF-MeOH extract. This process was modified from Sun et al. (1992) for isolating crude soluble polysaccharides of ginseng leaves.

A Frozen-thawed Bovine Luteal Cell System to Screen Luteotropic Activity in Drugs