Platycodon grandiflorus polysaccharide regulates colonic immunity through mesenteric lymphatic circulation to attenuate ulcerative colitis / 中国天然药物

Platycodon grandiflorus polysaccharide (PGP) is one of the main components of P. grandiflorus, but the mechanism of its anti-inflammatory effect has not been fully elucidated. The aim of this study was to evaluate the therapeutic effect of PGP on mice with dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) and explore the underlying mechanisms. The results showed that PGP treatment inhibited the weight loss of DSS-induced UC mice, increased colon length, and reduced DAI, spleen index, and pathological damage within the colon. PGP also reduced the levels of pro-inflammatory cytokines and inhibited the enhancement of oxidative stress and MPO activity. Meanwhile, PGP restored the levels of Th1, Th2, Th17, and Treg cell-related cytokines and transcription factors in the colon to regulate colonic immunity. Further studies revealed that PGP regulated the balance of colonic immune cells through mesenteric lymphatic circulation. Taken together, PGP exerts anti-inflammatory and anti-oxidant effect and regulates colonic immunity to attenuate DSS-induced UC through mesenteric lymphatic circulation.

Analysis and Contents Determination of Heavy Metals in Platycodon grandiflorus at Different Altitudes in the Three Gorges Reservoir Area / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of heavy metals in Platycodon grandiflorus, and to explore the effects of altitude on the contents of heavy metals. METHODS:The contents of As,Hg,Pb,Cu and Cd were determined by hydride generation,hydride generation method,graphite furnace method,flame spectrometry,graphite furnace meth-od. SPSS 19.0 software was adopted for clustering analysis and correlation analysis. RESULTS:The linear ranges of As,Hg,Pb, Cu and Cd were 0-16 μg/L(r=0.9960),0-18 μg/L(r=0.9996),0-50 μg/L(r=0.9997),0-0.8 μg/L(r=0.9990)and 0-4 μg/L (r=0.9980). Detection limit were 1.3×10-3,2.4×10-4,1.9×10-3,0.33,1.8×10-4 mg/kg. Quantitation limit were 4.4×10-3,8.0×10-4, 6.2 × 10-3,1.1,6.0 × 10-3 mg/kg. RSDs of precision,stability and repeatability tests were all lower than 3.0%. Recoveries were 99.1%-107.1%(RSD=3.13%,n=6),94.1%-100.5%(RSD=2.36%,n=6),98.9%-104.8%(RSD=1.81%,n=6),92.7%-100.3%(RSD=2.92%,n=6),96.6%-99.9%(RSD=1.26%,n=6). Samples with similar altitudes were clustered into a class;the con-tents of heavy metals decreased as the increase of altitude;there was significant correlation among 5 heavy metals. CONCLU-SIONS:The method is simple,precise,stable and repeatable,and can be used for simultaneous determination of heavy metals' contents in P. grandiflorus. The altitude has a certain effect on the quality of P. grandiflorus.

In vitro autotetraploid induction and analysis on DNA methylation diversity of Platycodon grandiflorum / 中国中药杂志

In order to investigate the epigenetic variations between diploid and autotetraploid of Platycodon grandiflorus. The diploid buds of P. grandiflorus were soaked in the mixture of different concentration colchicines and 0.002 g•mL ⁻¹ dimethyl sulphoxide (DMSO)．The identification of autotetraploid plants were based on morphological characteristics, chromosome number and flow cytometry. And then the level and pattern of DNA methylation explored by using the technology of methylation sensitive amplified polymorphism (MSAP)．The result demonstrated that the buds soaked in 0.2% colchicines and 0.002 g•mL ⁻¹ DMSO solution for 12 h was ideal conditions to induce autotetraploid of P. grandiflorus, with induction rate of 32.0%.The diploid and tetraploid plants existed distinctly differences in morphological indexes.Totally,1 586 bands were amplified by 20 pairs of selective primers, of which 764 and 822 bands were detected in diploid and autotetraploid respectively. The total methylation ratio,full methylation ratio and hemimethylated ratio were 91.25%,61.25% and 30.65% in diploid of P. grandiflorus,respectively.However,the total methylation ratio,full methylation ratio and hemimethylated ratio of autotetraploid of P. grandiflorus were 86.13%,54.38% and 31.75%, respectively. Compared with diploid, the genomic DNA total methylate ratio and full methylation ratio of autotetration plants decreased by 6.02% and 7.14%.But the hemimethylated ratio of autotetraploid was higher than that of diploid, which more than 1.6%. All this results indicated that DNA methylation patterns have adjusted during the polyploidy process．.

An HPLC-MS/MS method for the quantitative determination of platycodin D in rat plasma and its application to the pharmacokinetics of Platycodi Radix extract / 中国天然药物

AIMS@#To develop an HPLC-MS/MS method for the quantification of platycodin D (PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract (PRE) containing PD.@*METHOD@#Plasma samples were pretreated with solid-phase extraction using an Oasis® HLB SPE cartridge. Madecassoside was used as the internal standard (IS). Chromatographic separation was achieved on an ODS column (100 mm × 2.1 mm i.d., 3.5 μm) with a mobile phase consisting of acetonitrile/water (30 : 70, V/V) containing 0.1 mmol·L(-1) ammonium acetate at a flow rate of 0.25 mL·min(-1). The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization (ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside (IS), respectively.@*RESULTS@#The calibration curve was linear from 5 to 2 000 ng·mL(-1) (r(2) >0.99) with a lower limit of quantification (LLOQ) of 5 ng·mL(-1). The intra- and inter-day precision (relative standard deviation, RSD) values were below 15% and the accuracy (relative error, RE) was from -15% to +15% at three quality control (QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be (0.48 ± 0.19)% when administered PD, and to be (1.81 ± 0.89)% when administered PRE.@*CONCLUSION@#The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.

Biotransformation of cantharidin by cell suspension cultures of Platycodon grandiflorus / 中草药

Object To investigate the biotransformation of cantharidin by cell suspension cultures of Platycodon grandiflorus (Jacq ) A DC Methods Cantharidin was added into cell suspension cultures of P grandiflorus and incubated for another six days The culture supernatant was extracted with EtOAc and then subjected to silica gel chromatography Results Two products were isolated as a mixture in a molar ratio of about 2∶1 Their structures were identified on the basis of spectroscopic data as 1? OH cantharidin (Ⅱa) and 1? OH cantharidin (Ⅱb) The relative configuration was elucidated according to NOESY spectrum Conclusion The two products were new cantharidin derivatives