ABSTRACT
<p><b>OBJECTIVE</b>To investigate the serum protein targets of Qianggu Decoction (, QGD) on treating osteoporosis by the proteomics analysis using tandem mass tag (TMT) and liquid chromatographytandem mass spectrometry (LC-MS/MS).</p><p><b>METHODS</b>Twenty serum protein samples were recruited (10 patients with primary type I osteoporosis before and after QGD treatment) and the high abundance ratios protein was removed, two serum samples were extracted and labeled with TMT reagent. Then, mass spectrometric detection, identification of differentially expressed proteins and bioinformatics analysis of differentially expressed proteins were carried out.</p><p><b>RESULTS</b>A total of 60 proteins were identified, within a 99% confidence interval, to be differentially regulated of which, 34 proteins were up-regulated and 26 proteins were down-regulated. Differentially expressed proteins analyzed by Gene Ontology (GO) annotation mainly get involved in 12 different biological processes, 7 types of cellular components, and 6 kinds of molecular functions. Angiotensinogen (AGT), stromelysin-1 (MMP3), heparanase (HPSE) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were screened as candidate protein targets of QGD treatment, which were related to metabolic mechanism of bone remodeling and/or bone collagen of osteoporosis. By the utilization of the protein-protein interaction network analysis tool named STRING10.0, it showed that AGT, MMP3, HPSE and GAPDH were located in the key node of the protein-protein interactions network. Furthermore, AGT, MMP3, HPSE and GAPDH were found to be directly related to BMP, MAPK, Wnt, SMAD and tumor necrosis factor ligand superfamily member 11 (TNFSF11) families.</p><p><b>CONCLUSIONS</b>The proteomics analysis by using TMT combined with LC-MS/MS was a feasible method for screening the potential therapeutic targets associated with QGD treatment. It suggests that AGT, MMP3, HPSE and GAPDH may be candidate protein targets of QGD treatment which can be used as therapeutic effect monitor and early diagnosis of primary type I osteoporosis.</p>
ABSTRACT
OBJECTIVE:To study the technical conditions and parameters of DiaionHP-20 macroporous adsorption resin for the purification of the anti-PMOP(primary Type I osteoporosis) active fraction from Camellia semiserrata seed.METHODS: With the elution ratios and purity of total flavonoids and polyphenol from Camellia semiserrata as indexes,the technical parameters of DiaionHP-20 macroporous adsorption resin including the adsorption quantity,the loading sample,the elution concentration and volume of alcohol were investigated.RESULTS: The adsorption quantity of DiaionHP-20 macroporous resin on total flavonoids and polyphenol were 59.5 mg?g-1 and 23.9 mg?g-1 respectively.The optimal technical conditions were as follows:the quantity of drug was 10.5 times that of resin;water-soluble foreign substance was washed off with distilled water weighing 3 times the weight of resin,and total flavonoids and polyphenol were eluted using 25% ethanol weighing 13.5 times the weight of resin.CONCLUSION:DiaionHP-20 macroporous resin had good purification effects on total flavonoids and polyphenol from Camellia semiserrata seeds.The process is simple,low cost,and suitable for lavge-scale production.