| Banqiao radix codonopsis is dry root of balloonflower secco plant sichuan codonopsis pilosula (Codonopsis tangshen Oliv.), which is from hubei enshi and is the spleen yiqi good medicine. We have known that it is rich in tangshen polysaccharide, dangshen saponins, sterols kind, triterpene composition and a variety of trace elements(such as Fe, Mn, Zn, Se, etc) and human body essential amino acids,which is supplement qi and blood, nourishing spleen, embellish lung and Promote the secretion, nourishing buffets of curing body virtual,rank first of the "four big name party" list.I have Investigated phenol-sulfuric acid method to determine the content of the polysaccharide and Investigated the color conditions,the detected wavelength is489nm, the amount of phenol addition is1.5mL,the amount of Strong sulfuric acid addition is6.0mL,placing the polysaccharide at the room temperature for40min is the optimum determination conditions, At the same time I have investigated the methodology, The precision, stability and reproducibility, and the recovery RSD<2%,proved the determination method is stable and reliable. With water extraction and alcohol heavy law to extract tangshen polysaccharide, and through petroleum ether skimmed, ethanol remove small molecules and decolore the color,and through savage method to take off some protein I got the refined board party polysaccharide, it’s content is90.36%±1.12%, which provides stable and reliable planning for the follow-up of pharmacodynamics experiment.IEC-6cells are training in CO2incubator at37℃5%CO2after the arrival, three days ChuanDai once, I began to experimentat the13generation,I found that250ug/mL,500ug/mL,1000ug/mL,2000ug/mL,4000ug/mL dose group can obviously promote the IEC-6cell proliferation with MTT method, and analyse the absorbency of banqiao radix polysaccharide, finally, it was sure that the dosewhich banqiao radix polysaccharide accelerate the cell proliferation from low to high is250ug/mL,1000ug/mL,4000ug/mL; Put thegr owing logarithm cells in6hole training board, after growing to single cells, with small silicon cells blow blow single-celled layer gently,generating about3mm2round defect area,addicting4000ug/mL board party polysaccharide solution, addicting the same amount of completely medium in the control group, respectively take a picture after dosing0h,24h,48h,72h with microscope,it was found that4000ug/mL board party polysaccharide can obviously accelerate the ce ll transitions of IEC-6faster; with HE dyeing experiment, the cells after joining250ug/mL,1000ug/mL,4000ug/mL dose group of b oard party have the gland trend, but the control group did not have,which is said that banqiao radix polysaccharide group canaccelerate the differentiation of IEC-6cell;Homogenate the cells and medium with cells homogenate device,I have got the clear after cen trifugal to get ODC protein, with EILSA kit to detect the expressio n level,the results as the control group cells ODC content is (24.419±0.699)pg/mL,the low dose group is (29.419±0.941)pg/mL,the medi um dose group is (40.412±0.942) pg/mL,the high dose group is(45.2 71±0.620)pg/mL,the results show that the polysaccharide board can significantly raise the level of IEC-6cells ODC; with half quantitati ve RT-PCR method to detecte bird decarboxylation enzyme mRNA level,250ug/mL,1000ug/mL,4000ug/mL dose group of board party polysaccharides can raise the level of ODCmRNA and show dose dependent compared with the control group;experimental results showed that the board party polysaccharides can promote the prolife ration, differentiation of IEC-6cells, its mechanism may involve the ODC protein and ODCmRNA expression which banqiao radix polys accharides can promote the content and the expression. |
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