Possession Of Codonopsis Determination Of The Main Active Ingredient In Its Polysaccharide Leydig Cells

Recently,the increasing environmental pollution and pressure in modern life have given rise to the incidence of infertility.It is estimated that about 6 to 10 percent of persons at reproduction-age suffer from infertility in China. Reproductive health has become a hot subject for scientific research.The non-toxic side effect natural drugs which can adjust the male reproductive function are becoming an urgent need of human health.Codonpsis thalictrifolia Wall,belongs to the family Campanulaceae and species of Codonopsis.It is a common drug used by peoples lived in Tibet.According to"ZangYaoZhi"and"Jingzhu Bencao",Codonpsis thalictrifolia Wall,has the effects of reducing inflammation,repercussion,aphrodisiac and tonic,benefits to spleen and stomach,and nourishes one's vitality.It plays a certain role in treatment of infertility.However,the chemical constituents and pharmacological effects of Codonpsis thalictrifolia Wall,have not been reported at present.According to the researches of Codonpsis,the main active ingredients are polysaccharides and saponins.This study selected the Codonpsis thalictrifolia Wall,as the object. The contents of polysaccharides and saponins were determined,the extraction process of polysaccharides was optimized,and the effects of polysaccharides on Leydig cells were investigated in this study.Extracting technology of polysaccharides was optimized by single factor experiment and orthogonal experiment.The factors including extraction temperature,time,ratio of material to liquid and extract times were studied in the single factor experiment.The single factor experiment showed that when the extraction temperature was higher than 70℃,extraction time was about 4-6h, ratio of material to liquid was greater than 1:25,extraction times was up to 3 times,the yield of total polysaccharides changed slowly.The orthogonal experimental result showed that the impact factors queued as follows:extraction time＞ratio of material to liquid＞extraction temperature＞extract times;the highest yield of polysaccharide was 10.41%.The optimism extraction conditions of the orthogonal experiment were as follows:extracting with 30-fold water for 4h thrice at 70℃.Considering the orthogonal experiment results and experimental costs,optimization program for extractions were extracting with 30-fold water for 4h twice at 70℃.The polysaccharide and saponins in Codonpsis thalictrifolia Wall,were mearured for the first time.The contents of polysaccharides and saponins from Codonpsis thalictrifolia Wall.were compared with Codonopsis(pilosula franch.) Nnannf.used by Chinese traditional medicine in common.The results showed that the contents of polysaccharide in Codonpsis thalictrifolia Wall.and Codonopsis(pilosula franch.) Nnannf.were 24.32% and 18.25%,the contents of saponins were 3.02%and 2.48%,respectively. These indicat that the contents of polysaccharides and saponins from Codonpsis thalictrifolia Wall.are higher than that from Codonopsis(pilosula franch.) Nnannf.significantly. The isolated Leydig cells function was confirmed by trypan blue exclusion experiment and the experiment of relationship between cultural time and testosterone secretion.Trypan blue exclusion experiment result showed the live-rate was between 78%and 98%when the Leydig cells were cultured within 32 hours in vitr.The relationship between cultural time and testosterone secretion experiment result confirmed the basis of secretion of testosterone increased with culture time,existing a linear relationship(R=0.886,P＜0.01) between them.The growth rates tend to slow down after 4h.Therefore,the cultrural time of after experiments was fixed on 4h.When the polysaccharides concentration was 2mg/mL,Codonpsis thalictrifolia Wall.polysaccharides can significantly reduce the rat Leydig cells activity of SOD(P＜0.01),lower the content of MDA(P＜0.01) and enhance the activity of GSH-PX(P＜0.01).The results of this study proved the contents of polysaccharides and saponins from Codonpsis thalictrifolia Wall.were higher than those from Codonopsis(pilosula franch.) Nnannf..The polysaccharides of Codonpsis thalictrifolia Wall.can scaveng free radical to protect the Ledig cells to some extent.