Effect of Ten Ingredients from Chinese Herbal Medi
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RESEARCH ARTICELS
Effect of Ten Ingredients from Chinese Herbal Medicine
on Proliferation of Chicken Embryo Fibroblast*
Yuanliang Hu**, Xiangfeng Kong, Deyun Wang, Jiaguo Liu, Baokang Zhang
Abstract: Ten Chinese herbal medicinal ingredients, astragalus polysaccharide (APS), Chinese angelica polysaccharide (CAPS), epimedium polysaccharide (EPS), isatis root polysaccharide (IRPS), propolis polysaccharide (PPS), astragalus flavone (AF), epimedium flavone (EF), propolis flavone (PF), astragalus saponin (AS) and ginsenoside (GS), were extracted and diluted into five concentrations which were incubated respectively with chicken embryo fibroblast (CEF) together in 96-well plate. At 24, 36, 48, 60 and 72 hours after cultivation, the proliferation of CEF was determined by MTT methods. The results showed that all of the ten could stimulate multiplication of the cell at suitable concentration and certain time points and the effects have relations with the dose and time.
Key words: Chinese herbal medicinal ingredient (CHMI); chicken embryo fibroblast (CEF); cell proliferation
Ginseng, Astragalus root, Chinese angelica root, Epimedium and Isatis root，Propolis are ordinary traditional Chinese medicine (TCM) respectively belonging to tonics and heat-clearing remedies [1]. Many researches have confirmed that many components from TCM possess immuno-enhancement and antiviral effects [2-5]. In the previous researches, the author extracted ten kinds of Chinese herbal medicinal ingredients (CHMI), astragalus polysaccharide (APS), Chinese angelica polysaccharide (CAPS), epimedium polysaccharide (EPS), isatis root polysaccharide (IRPS), propolis polysaccharide (PPS), astragalus flavone (AF), epimedium flavone (EF), propolis flavone (PF), astragalus saponin (AS) and ginsenoside (GS), from above-mentioned TCM and determined the safe concentration for each ingredient on chicken embryo fibroblast (CEF)[6]. In this research their effects on proliferation of CEF were further determined in order to evaluate them comprehensively and select better ones for development of new immuno-potentiator.
MATERIALS AND METHODS
1. Preparation of CHMI
APS, CAPS, EPS, IRPS and PPS, whose extractions were by the means of water-decocting and alcohol-sedimentation and these contents determination were by sulfuric acid anthrone method. AF and EF were extracted by polyamide chromatography, PF, by acid-base precipitation, and the three contents were determined by rutin method. AS and GS, whose extractions were by wide absorbed resin chromatography and content determinations were by TLC spectrophotometry. According to the determining results of safe concentration on CEF [6], the ten CHMI were diluted into five different concentrations, autoclaved and stored at 4℃.
2. Reagents
MEM medium, Gibco BRL, was supplemented with L-glutamine up to 0.03%, penicillin, 100IU/ml, streptomycin, 100μg/ml, and fetal calf serum, 5% (as growth solution) or 2% (as maintain solution) ......
RESEARCH ARTICELS
Effect of Ten Ingredients from Chinese Herbal Medicine
on Proliferation of Chicken Embryo Fibroblast*
Yuanliang Hu**, Xiangfeng Kong, Deyun Wang, Jiaguo Liu, Baokang Zhang
Abstract: Ten Chinese herbal medicinal ingredients, astragalus polysaccharide (APS), Chinese angelica polysaccharide (CAPS), epimedium polysaccharide (EPS), isatis root polysaccharide (IRPS), propolis polysaccharide (PPS), astragalus flavone (AF), epimedium flavone (EF), propolis flavone (PF), astragalus saponin (AS) and ginsenoside (GS), were extracted and diluted into five concentrations which were incubated respectively with chicken embryo fibroblast (CEF) together in 96-well plate. At 24, 36, 48, 60 and 72 hours after cultivation, the proliferation of CEF was determined by MTT methods. The results showed that all of the ten could stimulate multiplication of the cell at suitable concentration and certain time points and the effects have relations with the dose and time.
Key words: Chinese herbal medicinal ingredient (CHMI); chicken embryo fibroblast (CEF); cell proliferation
Ginseng, Astragalus root, Chinese angelica root, Epimedium and Isatis root，Propolis are ordinary traditional Chinese medicine (TCM) respectively belonging to tonics and heat-clearing remedies [1]. Many researches have confirmed that many components from TCM possess immuno-enhancement and antiviral effects [2-5]. In the previous researches, the author extracted ten kinds of Chinese herbal medicinal ingredients (CHMI), astragalus polysaccharide (APS), Chinese angelica polysaccharide (CAPS), epimedium polysaccharide (EPS), isatis root polysaccharide (IRPS), propolis polysaccharide (PPS), astragalus flavone (AF), epimedium flavone (EF), propolis flavone (PF), astragalus saponin (AS) and ginsenoside (GS), from above-mentioned TCM and determined the safe concentration for each ingredient on chicken embryo fibroblast (CEF)[6]. In this research their effects on proliferation of CEF were further determined in order to evaluate them comprehensively and select better ones for development of new immuno-potentiator.
MATERIALS AND METHODS
1. Preparation of CHMI
APS, CAPS, EPS, IRPS and PPS, whose extractions were by the means of water-decocting and alcohol-sedimentation and these contents determination were by sulfuric acid anthrone method. AF and EF were extracted by polyamide chromatography, PF, by acid-base precipitation, and the three contents were determined by rutin method. AS and GS, whose extractions were by wide absorbed resin chromatography and content determinations were by TLC spectrophotometry. According to the determining results of safe concentration on CEF [6], the ten CHMI were diluted into five different concentrations, autoclaved and stored at 4℃.
2. Reagents
MEM medium, Gibco BRL, was supplemented with L-glutamine up to 0.03%, penicillin, 100IU/ml, streptomycin, 100μg/ml, and fetal calf serum, 5% (as growth solution) or 2% (as maintain solution) ......
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