Investigate On The Effervescent Granules Prepared Using Eucommia Ulmoides Oliv. And Phyllostachys Pubescens Leaves

Chlorogenic acids from E. ulmoides Oliv. leaves is a kind of polyphenolic compound, which has been reported to have antidiabetic, antihypertensive and hypolipidemic activities, part from other biological properties. Polysaccharides from P. pubescens leaves is a kind of heteropolysaccharide with medium relative molecular mass. Recent studies have shown that polysaccharides sourced from P. pubescens leaves have antidiabetic properties. Effervescent granule is an ideal form for preparing a natural antidiabetic agent, for it is good taste, convenient to eat and easy to carry and store. Therefore, we prepared effervescent granules with 5% each of polysaccharides and chlorogenic acids from P. pubescens and E. ulmoides Oliv. leaves (EBEG). The effective ingredients of EBEG were determined, while the hypoglycemic activity and cytotoxicity of EBEG were measured by MTS method and a glucose assay kit in HepG2 and HUVEC cells. Research results and analysis: 1.The preparation of EBEGEBEG were prepared using acid-base mixing granulation method instructed by natural pharmaceutical chemistry theory.According to single factor experiment, the optimum proportion of acid and alkali was 5:4. The qualification of effervescent granules were estimated in terms of melting time. The compound of effervescent granules had been studied by orthogonal experiment. We choose mannitol as filler, aspartme:acesulfame (1:1) as flavoring agent, citric acid:tartaric acid (1:1) as acid source and sodium bicarbonate wrapped up by 4% PEG6000 as alkali source, and the dosage of them were 45.5%,0.5%,25% and 20%, respectively.The results showed that the process of the preparation of EBEG conform to the requirements of pharmacopoeia. 2.Determination of effective ingredients of EBEGEBEG was prepared using polysaccharides and chlorogenic acids from P. pubescens and E. ulmoides Oliv. leaves and the effective ingredients of it were determined by HPLC and vitriol-phenol Colorimetry methods. The quality and contents of the effective ingredients were examined in according with the regulations of "Chinese Pharmacopoeia".The chlorogenic acid in EBEG was identified by TLC. The results showed that the positon of the color spots of EBEG was the same as that of control, while the negative control had no interference.The results are agreement with those reported in the literature, as these phytochemicals presenting RSD values below 5%.The results showed that the methods of quality control were reproducibility and reliability. 3.The hypoglycemic activity of EBEGThe hypoglycemic activity of EBEG were measured by the glucose assay kit in HepG2 cells.The hypoglycemic effect of EBEG was much greater than that of granules prepared using either component alone. The uptake of glucose of EBEG increased 2 %～24% more than that of effervescent granules containing only polysaccharides from P. pubescens leaves, while it increased 13%～28% more than that of effervescetn granules chlorogenic acids from E. ulmoides Oliv. leaves. Furthermore, at a concentration of 200 μg/mL, EBEG exhibited a glucose uptake of 156.35%.The results showed that the individual components of EBEG acted synergistically to produce a greater hypoglycemic effect. 4.The cytotoxicity evaluation of EBEGThe cytotoxicity of EBEG were evaluated in HepG2 and HUVEC cells, and the cell growth curves within 7 days were measured by MTS method.The granules prepared using either component alone inhibited the cells proliferation, while EBEG exerted no significant effect either on the cells growth or on cells proliferation. Furthermore, the cell viability and growth curve of EBEG were similar to those of untreated controls.Though the HepG2 cells viability of EBEG showed dose-dependent, the cytotoxocity of EBEG exerted least influences, as it did at the concentration of 200 μg/mL that the cells viability was 99.82%. For HUVEC cells, EBEG did not show any effect on the cells viability. At low concentration, the cells viability was 105.45%, for EBEG could promote the cells growth and proliferation.The results showed that EBEG has potential for use as a natural health product to combat diabetes and exert a less toxic effect on cells, for polysaccharides from P. pubescens leaves and chlorogenic acids from E. ulmoides Oliv. leaves reduce the cytotoxicity synergistically.