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Yang B.,Hunan University | Li Z.-J.,Hunan University | Deng Z.,Hunan University | Wu G.-B.,Hunan Academy of Chinese Medicine
Chinese Journal of Tissue Engineering Research

Background: Duhuojisheng decoction is a basic prescription of Chinese medicine for treatment of lumbar disc herniation, but its mechanism is yet unclear. Objective: To study the effect of Duhuojisheng decoction on prostaglandin E2 in rabbits with lumbar disc herniation. Methods: Models of lumbar disc herniation were established in New Zealand rabbits using molding device, and verified successfully through CT examination at 1 week after modeling. The model rabbits were randomly divided into model group (treated with normal saline), voltaren group (treated with 2.3 mg/kg voltaren) and Duhuojisheng decoction group (treated with 2.3 mg/kg Duhuojisheng decoction). The treatment was done twice a day, continued 2 weeks. Normal rabbits and rabbits undergoing sham operation were used as controls.nucleus pulpous and annulus fibrosus tissue visible Results and Conclusion: {circled digit one}Hematoxylin-eosin staining results showed that granulation tissue ingrowth, neovascularization and massive infiltration of inflammatory cells were visible in nucleus pulpous and annulus fibrosus tissue after successful modeling of lumbar disc herniation. However, there was a remarkable relief in inflammatory cell infiltration after intragastric administration of Duhuojisheng decoction and voltaren. {circled digit two}The results of enzyme linked immunosorbent assay showed that prostaglandin E2 levels in local soft tissues and peripheral plasma were increased significantly after modeling, and then decreased afte administration of Duhuojisheng decoction and voltaren (P < 0.01). However, there was no difference in prostaglandin E2 levels after administration of Duhuojisheng decoction and voltaren (P > 0.05). These findings indicate that prostaglandin E2 plays a critical role in lumbar disc herniation, mediating in vivo inflammatory reaction. Duhuojisheng decoction can cure lumbar disc herniation through regulation of prostaglandin E2 expression, and its anti-inflammatory mechanism is similar to nonsteroidal anti-inflammatory drugs, such as voltaren. Source

Hu X.,Central South University | Chen L.,Hunan Academy of Chinese Medicine | Shi S.,Central South University | Cai P.,Hunan Academy of Chinese Medicine | And 2 more authors.
Journal of Pharmaceutical and Biomedical Analysis

Lonicerae macranthoides with strong antioxidant activity is commonly used in traditional Chinese medicine and folk tea/beverage. However, detailed information about its antioxidant activity and bioactive compounds is limited. Then at first, we comparatively evaluated total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities of water extract, petroleum ether, ethyl acetate and n-butanol fractions of L. macranthoides. Ethyl acetate fraction exhibited the highest level of TPC (207.38 mg GAE/g DW), TFC (53.06 mg RE/g DW) and the best DPPH scavenge activity and reducing power. n-Butanol fraction showed the best ABTS+ and O2 - scavenging activities. Interestingly, water extract, ethyl acetate and n-butanol fractions showed stronger antioxidant activities than positive control, butylated hydroxytoluene (BHT). After that, thirty-one antioxidant phenolic compounds, including twenty-two phenolic acids and nine flavonoids, were screened by DPPH-HPLC experiment and then identified using HPLC-DAD-QTOF-MS/MS. It is noted that twenty-one compounds (1, 3-4, 6-17, 19, 23, 26, 28-29, and 31), as far as was known, were discovered from L. macranthoide for the first time, and eleven of them (3-4, 10-17, and 23) were reported in Lonicera species for the first time. Results indicated that L. macranthoides could serve as promising source of rich antioxidants in foods, beverages and medicines for health promotion. © 2016. Source

Zhang S.-H.,Hunan Academy of Chinese Medicine | Hu X.,Central South University | Shi S.-Y.,Central South University | Huang L.-Q.,Chinese Academy of Sciences | And 3 more authors.
Analytical and Bioanalytical Chemistry

A major challenge of profiling chlorogenic acids (CGA) in natural products is to effectively detect unknown or minor isomeric compounds. Here, we developed an effective strategy, typical ultraviolet (UV) spectra in combination with diagnostic mass fragmentation analysis based on HPLC–DAD–QTOF-MS/MS, to comprehensively profile CGA in the buds of Lonicera macranthoides. First, three CGA UV patterns were obtained by UV spectra screening. Second, 13 types of CGA classified by molecular weights were found by thorough analysis of CGA peaks using high-resolution MS. Third, selected ion monitoring (SIM) was carried out for each type of CGA to avoid overlooking of minor ones. Fourth, MS/MS spectra of each CGA were investigated. Then 70 CGA were identified by matching their UV spectra, accurate mass signals and fragmentation patterns with standards or previously reported compounds, including six caffeoylquinic acids (CQA), six diCQA, one triCQA, three caffeoylshikimic acids (CSA), six diCSA, one triCSA, three p-coumaroylquinic acids (pCoQA), four p-coumaroylcaffeoylquinic acids (pCoCQA), four feruloylquinic acids (FQA), five methyl caffeoylquinates (MCQ), three ethyl caffeoylquinates (ECQ), three dimethoxycinnamoylquinic acids (DQA), six caffeoylferuloylquinic acids (CFQA), six methyl dicaffeoylquinates (MdiCQ), four FQA glycosides (FQAG), six MCQ glycosides (MCQG), and three ethyl dicaffeoylquinates (EdiCQ). Forty-five of them were discovered from Lonicera species for the first time, and it is noted that CGA profiles were investigated for the first time in L. macranthoides. Results indicated that the developed method was a useful approach to explore unknown and minor isomeric compounds from complex natural products. [Figure not available: see fulltext.] © 2016 Springer-Verlag Berlin Heidelberg Source

Liang X.,Hunan Academy of Chinese Medicine | Zhang Y.,Central South University | Chen W.,Hunan Academy of Chinese Medicine | Cai P.,Hunan Academy of Chinese Medicine | And 3 more authors.
Journal of Chromatography A

A challenge in coupling high-speed counter-current chromatography (HSCCC) online with high performance liquid chromatography (HPLC) for purity analysis was their time incompatibility. Consequently, HSCCC-HPLC was conducted by either controlling HPLC analysis time and HSCCC flow rate or using stop-and-go scheme. For natural products containing compounds with a wide range of polarities, the former would optimize experimental conditions, while the latter required more time. Here, a novel HSCCC-HPLC-diode array detector-mass spectrometry (HSCCC-HPLC-DAD-MS) was developed for undisrupted purification, analysis and identification of multi-compounds from natural products. Two six-port injection valves and a six-port switching valve were used as interface for collecting key HSCCC effluents alternatively for HPLC-DAD-MS analysis and identification. The ethyl acetate extract of Malus doumeri was performed on the hyphenated system to verify its efficacy. Five main flavonoids, 3-hydroxyphloridzin (1), phloridzin (2), 4',6'-dihydroxyhydrochalcone-2'- O-β- d-glucopyranoside (3, first found in M. doumeri), phloretin (4), and chrysin (5), were purified with purities over 99% by extrusion elution and/or stepwise elution mode in two-step HSCCC, and 25. mM ammonium acetate solution was selected instead of water to depress emulsification in the first HSCCC. The online system shortened manipulation time largely compared with off-line analysis procedure and stop-and-go scheme. The results indicated that the present method could serve as a simple, rapid and effective way to achieve target compounds with high purity from natural products. © 2015 Elsevier B.V. Source

Peng M.-J.,JiShou University | Shi S.-Y.,Central South University | Chen L.,Hunan Academy of Chinese Medicine | Zhang S.-H.,Hunan Academy of Chinese Medicine | And 2 more authors.
Analytical and Bioanalytical Chemistry

Screening and analysis of bioactive compounds from natural products is challenging work due to their complexity. This study presents the first report on hyphenation of solid-phase ligand-fishing using immobilized xanthine oxidase microcolumn (IXOM) and high-performance liquid chromatography–diode array detector–tandem mass spectrometry (HPLC–DAD–MS/MS) for screening and identification of XO inhibitors from complex mixtures. Solid-phase ligand-fishing system was hyphenated with the HPLC system via four-port switching valve and a six-port injection valve as an interface for transferring effluents from IXOM to HPLC, and collecting chromatograms from LFMC (ligand-fishing microextraction column) and C18 column in a run by only one DAD. Mixtures containing allopurinol (positive control) and tryptophane (negative control) were analyzed in order to verify the specificity and reproducibility of the approach. Subsequently, the newly developed system was applied to screening and identification of XO inhibitors from L. macranthoides and its human microsomal metabolites. Six prototype compounds (3-caffeoylquinic acid, 5-caffeoylquinic acid, 4-caffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid) and three metabolites (3-caffeoyl-epi-quinic acid, 5-caffeoyl-epi-quinic acid, 4-caffeoyl-epi-quinic acid) with XO binding affinities were identified. The XO inhibition activities of six prototype compounds were evaluated and confirmed using in vitro enzymatic assay. With the online system developed here, we present a feasible, selective, and effective strategy for rapid screening and identification of enzyme inhibitors from complex mixtures. © 2016 Springer-Verlag Berlin Heidelberg Source

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