Effects of Root Extracts from Panax ginseng C. A. Meyer (Araliaceae) of Different Ages on K562 Cells

Xunan Lyu, Yizhen Zhang, Qing Mu, Zhou Cheng, Jiakuan Chen, Wenju Zhang

Ministry of Education, Key Laboratory for Biodiversity Science and Ecological Engineering, Institute of Biodiversity Science, School of Life Sciences, Fudan University, Shanghai, China.
Ministry of Education, Key Laboratory for Biodiversity Science and Ecological Engineering, Institute of Biodiversity Science, School of Life Sciences, Fudan University, Shanghai, China;.
School of Life Science and Technology, Tongji University, Shanghai, China..
School of Pharmacy, Fudan University, Shanghai, China;.

DOI: 10.4236/ajps.2013.46160                

Abstract

It is well accepted in China that elder ginsengs have more bioactivity and value than younger ones. However, there is little research about the comparison of beneficial effects of ginsengs with different ages. In this study, ginseng root extracts (GRE) were extracted from ginsengs of 5, 8, 12, 14, and 16 years old, respectively, using 55% ethanol and their effects on human leukemic K562 cells within 48 hours were tested by using Cell Counting Kit-8. The results show that there are significant increases in the cell viability of all the GRE groups compared with Control group within 32 hours. Furthermore, the growth curves of GRE groups were obviously distinct from each other. The cell viability of 5-year-old and 8-year-old GRE groups kept a rapid increase while that of 16-year-old GRE group showed a strong fluctuation within 28 hours. Our results demonstrate that root extracts from ginsengs of different ages contain different bioactivity constituents and have different effects on cell.

Keywords

Panax ginseng; Root Extracts; Ages; K562 Cell Line

1. Introduction

Panax ginseng C. A. Meyer (ginseng) is a famous herbal medicine with a wide range of therapeutic benefits, such as inhibiting tumor growth, regulating immune system, inducing the cell differentiation, etc. [1,2]. It is well accepted that elder ginsengs have more bioactivity and value than younger ones based on Chinese conventional concept; however, there is little research about the comparison of beneficial effects of ginsengs with different ages.

K562 cell line was established from a patient with chronic myelogenous leukemia and has been widely used both in vitro experiments at present [3]. Ginsenosides and polysaccharides are regarded as the primary active ingredients of ginseng [1,2]. Total ginsenosides of ginseng could not only inhibit the proliferation of K562 cells but also induce the differentiation by inhibiting the expression of erythropoietin receptor protein [4,5]. Moreover, studies have provided clear evidence that polysaccharides from Ginseng also exert antiproliferative activeity against K562 cells at a certain concentration range in vitro experiment [6,7]. Except for ginsenosides and polysaccharides, ginseng roots also contain some other constituents such as peptides, polyacetylenic alcohols, flavones, and fatty acids which also contribute to a wide range of beneficial effects of ginseng [1,2]. However, the general effects of ginsengroot extracts (GRE) on K562 cells have not been previously investigated.

The purpose of this study was to compare the effects of GRE of different ages on K562 cell line.

2. Materials and Methods

2.1. Sample Collection

Roots of ginsengwere collected from Huairen and Ji’an, Jilin province, China respectively (shown in Table 1). These ginsengs grew in the same cultivated condition. Roots of ginseng were dried at 50˚C and were preserved at –20˚C. Each group consists of 5 individuals except H16 group (4 individuals).

2.2. Root Extracts of Ginsengs

Ginseng samples of different ages were dried at 50˚ Cand crushed (40 mesh sieve), respectively. Each above sample was accurately weighed 6.000 g and 18.2 mL/g of 55% ethanol was added. After soaking 12 hours at room temperature, the sample suspension was extracted under ultrasonic for 42 min, then centrifuged and the supernatant was collected. The extraction was repeated four times. The supernatants were merged and evaporated to dryness under vacuum at temperature 45˚C. The dried ginseng extracts were accurately weighed to calculate the contents of total GRE and used as pharmacological experiments. A stock solution was prepared by dissolving GRE in ddH₂O and stored at –20˚C. To analysis amount in different polarity, the extract resin was resuspended in 30 mL distilled water and then partitioned with chloroform (30 mL × 3). The chloroform extracts were merged together and dried in a vacuum at 45˚C and weighed accuratelyto calculate the contents, then stored at 4˚C. The above remains of the ginseng extracts aqueous were then extracted with butanol (30 mL × 3). The final remains of the ginseng extracts aqueous were removed to dry and weighed accuratelyto calculate the contents.

2.3. Cell Culture and Cell Growth Curve Detection

K562 human leukemia cell line was obtained from Shanghai Institute of Cell, Chinese Academy of Sciences. Cells were cultured in RPMI-1640 medium (Gibco-BRL, Gaithersburg, MD) supplemented with 10% fetal bovine serum (Gibco-BRL), 100 μg/ml streptomycin (Sigma Chemical Co., St. Louis, MD) and 100 IU/ml penicillin (Sigma) at 37˚C. Cells were seeded at 1 × 10⁵ cells/well in 96-well plate for 48 hours with or without a final concentration of 1 mg/ml GRE named Control group, H5 (5-year-old) group, H8 (8-year-old) group, H12 (12-yearold) group, H14 (14-year-old) group and H16 (16-yearold) group respectively. Each group had 6 repeats. Cell viability was analyzed by Cell Counting Kit-8 (CCK-8, Dojindo, Kumamoto, Japan) every 4 hours within 48 hours.

2.4. Statistical Analysis

Results were expressed as the mean ± standard deviations (SD). The effect on each parameter was examined by one-way analysis of variance (ANOVA) and independent-sample t test. P values less than 0.05 were considered statistically significant.

3. Results

3.1. Content of Total GRE and Their Constituents

Table 1 shows contents of total GRE in ginseng of different age groups. H5 group (5-year-old) has the lowest content (30.87%) and H16 group (16-year-old) has the highest (46.43%). Contents of ginsenosides and polysaccharides in GRE were measured and results were showed in table. H16 group has the highest content of ginsenosides in GRE (16.11%) while H5 has the highest content ofpolysaccharides (56.37%).

Conflicts of Interest

The authors declare no conflicts of interest.

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