Ginseng polysaccharides on HCG-induced luteal cell function of the particle
Author: Sun Yan, Feng Li, Hong-Jun Wang, the
spring-chi, Chu Zheng, Qiang Wang, Zhang Zhiqiang, Wu Yimin
【Abstract】 Objective To study the ginseng polysaccharides
on the role of ovarian function in rats. Methods Cell culture in vitro observation of ginseng polysaccharides on HCG-induced
luteal progesterone secretion in granulosa cells and the effects with cAMP and examined the survival rate of oocytes.
Isolated and cultured luteal, particles and oocytes, as control group with different concentrations of ginseng polysaccharide experimental group were measured using radioimmunoassay of progesterone and cAMP generated content, cell viability detected by MTT method. Results Compared with control group, progesterone content: luteal cells (6 573.46 ?
185.14) and (4509.55 ? 126.82) pmol, granule cells (197.16 ?
42.87) and (320.42 ? 12.46) pmol. cAMP: luteal cells (31.20 ? 17.13) and (65.26 ? 15.93) fmol, granule cells (121.15 ?
19.96) and (296.42 ? 27.28) fmol. Conclusion ginseng
polysaccharides inhibit HCG-induced progesterone secretion of luteal cells to promote the HCG-induced progesterone secretion in granulosa cells. The luteal cells together with the
granular cell cAMP generation, ginseng polysaccharides so that the rate of oocyte growth inhibition lower interval was dose-
Key words Panax ginseng polysaccharide; progesterone; cyclic adenosine monophosphate; human chorionic gonadotropin
The Effect of Ginseng Polysugar on Secretion Function of Luteal and Granulose Cell of Rat in vitro
Abstract: ObjectiveTo study using of ginseng polysugar on secretion function of female rat ovary. The experiment had observed progesterone and cAMP by HCG inducing luteal and
granulose cell as well as living cell rate of oocyte in vitro.MethodsRat luteal and granulose cell and oocyte was cultivated , cell counting was worked by microscope, progesterone and cAMP content was determined by
radioimmunoassay kit (RIA kit). Cell exist rate of oocyte was determined by MTT way. ResultsProgesterone: the luteal cell test groups compare with control group, 6573.46 ? 185.14 and
( 4509.55 ? 126.82) pmol.The granulose cell test groups compare with control group, 197.16 ? 42.87 and (320.42 ?
12.46) pmol.cAMP: the luteal cell test groups compare with control group, 31.20 ? 17.13and (65.26 ? 15.93) fmol, the
granulose cell test groups compare with control group, 121.15 ? 19.96 and (296.42 ? 27.28) fmol.ConclusionThis treatise
had demonstrated that ginseng polysugar can inhibit HCG inducing progesterone secretion and enhancing cAMP content of luteal and granulose cell of rat, ginseng polysugar has lower oocytes cell grow up inhibitory rate. It is evident that ginseng polysugar possesses characteristic of dose dependent.
Key words: Ginseng polysugar; Progesterone; cAMP; HCG
Many studies have shown that ginseng polysaccharides have a nourishing vitality, enhance the role of gonadal function , its adoption under the influence of the hypothalamus pituitary gonadal axis secretion system, the target organs and tissues through the cAMP intracellular information transmission system to start a series of physiological and biochemical reaction, ovary, as the hypothalamus - pituitary -
gonadal axis role in the reproductive system, its subject the role of ginseng polysaccharides to accelerate the process of sexual maturation and reproductive cycle also occurs in different functional changes. Ginseng polysaccharides on
reproductive cells more complicated, yet lack of a comprehensive system, detailed reports, in order to explore the ginseng polysaccharides on human chorionic gonadotropin (HCG)-induced rat luteal and granulosa cells function, this study used in vitro culture methods observed ginseng
polysaccharides on HCG-induced rat corpus luteum and granulosa cells secreted progesterone and cAMP content changes and cell
viability were detected. The content of this study no relevant reports, hereby reports the following results.
1.1 Animals and cells in body weight of about 250 ~ 300 g adult female rats were purchased from the Experimental Animal Center, China Medical University, in this laboratory sub-cage
farming, grain feed, automatic water, to adapt to 1 week after the experiment. Luteal cells, granule cells and oocytes from rat ovary.
1.2 Reagent and instrument MoCoy's 5a medium dry, collagenase, progesterone, human chorionic gonadotropin and heparin were purchased from Sigma Company; calf serum were
purchased from Shanghai Biological Engineering Company; pregnant mare serum were purchased from Experimental Animal Center of Tianjin ; ginseng polysaccharide Chemistry, Jilin University teaching and research benefit from the gift (2 g /
L); 3H-progesterone were purchased from Shanghai Institute of Biological Products; 125I-cAMP RIA kit were purchased from
Shanghai Traditional Chinese Medicine, xylene, PPO, POPOP, HCI , NaOH, etc. purchased from Shenyang Chemical Reagent Company, were analytically pure; CO2 Incubator: The United States Shel-Lab Inc., model 1815TC; Clean Bench: Jiangsu Suzhou Antai Air Technology Co., Ltd., model SW-CJ-2FD;
inverted phase contrast Microscope: Japan Olympus Corporation, model CX21FS1. BECKMAN: American Beckman Company, model J2-21;
liquid scintillation counter: Sweden LKB Company (machine efficiency 50%), model LKB1211; Kontro γ - Counter: Swiss
company, model Kontro-28D.
2.1 Cell Culture
2.1.1 Preparation of luteal cells and cultivation of the
female rats were injected subcutaneously neck, pregnant mare serum 50U / only, 48 h after the subcutaneous injection of human chorionic gonadotropin 45U / only, 1 week later, animals were decapitated, sterile take ovaries, the corpus luteum of
ovary Shredded, 0.2% collagenase 5 ml digestion at 37 ?
incubator every 15 min straw dogs new tricks, 1 h, add 5 ml MoCoy's 5a medium, blending, 200 r / min , centrifugation 2 min. Abandoned precipitation, 1300 r / min, centrifuge 10 min,
discard supernatant, add culture medium 10 ml, 1 300 r / min, centrifuge 10 min, washed 2 times. Add a new medium, blending, take 50 μl, together with 50 μl 0.4% trypan blue, cell
counting, calculating the percentage of cells in the total number of living cells. With 5% fetal calf serum MoCoy's 5a medium raised to 300,000 cells / 0.5 ml, by adding ginseng polysaccharide (30 ng / ml), inoculated 24-well plate, 5% CO2,
95% ventilation, 37 ? cultured 24 h, 0.5 ml serum-free
MoCoy's 5a medium wash one time, plus ginseng polysaccharides
containing 0.9 ~ 1 ml serum-free culture medium, 24 h later,
for fluid, washed 1 time. Serum-free culture medium plus 1 ml,
as control group and experimental group ginseng
polysaccharide, ginseng polysaccharides containing cell
culture medium 48 h, collected culture medium, boil 5 min, -
20 ? storage, for progesterone and cAMP radioimmunoassay.
2.1.2 Preparation of granulosa cells and cultivation of the female rat abdominal subcutaneous injection of dehydroepiandrosterone diethylstilbestrol oil 0.5 mg/0.1 ml / only, continuous injection of 4 d, 5 days, animals were decapitated, sterile taken ovary, will be used dehydrogenase diethylstilbestrol oil deal with the trumpet of the rat ovarian needle (41 / 2) punctured follicles and release
granule cells. Centrifugal, closed cell, plus an appropriate culture medium, take 50 μl, add 50 μl 0.4% trypan blue, and