Specific DNA barcodes screening, germplasm resource identification, and genetic diversity analysis of Platycodon grandiflorum / 药学学报

Platycodonis Radix is the dry root of Platycodon grandiflorum of Campanulaceae, which has a variety of pharmacological effects and is a commonly used bulk Chinese medicine. In this study, the chloroplast genome sequences of six P. grandiflorum from different producing areas has been sequenced with Illumina HiSeq X Ten platform. The specific DNA barcodes were screened, and the germplasm resources and genetic diversity were analyzed according to the specific barcodes. The total length of the chloroplast genome of 6 P. grandiflorum samples was 172 260-172 275 bp, and all chloroplast genomes showed a typical circular tetrad structure and encoded 141 genes. The comparative genomics analysis and results of amplification efficiency demonstrated that trnG-UCC and ndhG_ndhF were the potential specific DNA barcodes for identification the germplasm resources of P. grandiflorum. A total of 305 P. grandiflorum samples were collected from 15 production areas in 9 provinces, for which the fragments of trnG-UCC and ndhG_ndhF were amplificated and the sequences were analyzed. The results showed that trnG-UCC and ndhG_ndhF have 5 and 11 mutation sites, respectively, and 5 and 7 haplotypes were identified, respectively. The combined analysis of the two sequences formed 13 haplotypes (named Hap1-Hap13), and Hap4 is the main genotype, followed by Hap1. The unique haplotypes possessed by the three producing areas can be used as DNA molecular tags in this area to distinguish from the germplasm resources of P. grandiflorum from other areas. The haplotype diversity, nucleotide diversity and genetic distance were 0.94, 4.79×10-3 and 0.000 0-0.020 3, respectively, suggesting that the genetic diversity was abundant and intraspecific kinship was relatively close. This study laid a foundation for the identification of P. grandiflorum, the protection and utilization of germplasm resources, and molecular breeding.

Prokaryotic expression, subcellular localization and enzymatic activity analysis of DXS gene from Platycodon grandiflorum / 药学学报

1-Deoxy-D-xylulose-5-phosphate synthase (DXS), the first key enzyme in 2-methyl-D-erythritol-4-phosphate (MEP) pathway, catalyzes the condensation of glyceraldehyde-3-phosphate with pyruvate to 1-deoxy-xylose-5-phosphate (DXP). In this study, PgDXS1, PgDXS2, and PgDXS3 genes were cloned from the root of Platycodon grandiflorum (P. grandiflorum). The open reading frame (ORF) of PgDXS1, PgDXS2, and PgDXS3 were 2 160, 2 208, and 2 151 bp in full length, encoding 719, 735, and 716 amino acids, respectively. Homologous alignment results showed a high identity of PgDXSs with DXS in Hevea brasiliensis, Datura stramonium and Stevia rebaudiana. The recombinant expression plasmids of pET-28a-PgDXSs were constructed and transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and the induced proteins were successfully expressed. Subcellular localization results showed that PgDXS1 and PgDXS2 were mainly located in chloroplasts, and PgDXS3 was located in chloroplasts, nucleus and cytoplasm. The expression of three DXS genes in different tissues of two producing areas of P. grandiflorum were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves of P. grandiflorum from Taihe. Under methyl jasmonate (MeJA) treatment, the expression levels of three PgDXS genes showed a trend of first decreasing and then increasing at different time points (3 - 48 h), and the activity of DXS showed a trend of first increasing and then decreasing in three tissues of P. grandiflorum. This study provides a reference for further elucidating the biological function of PgDXS in terpenoid synthesis pathway in P. grandiflorum.

Polygalacin D inhibits the growth of hepatocellular carcinoma cells through BNIP3L-mediated mitophagy and endogenous apoptosis pathways / 中国天然药物

Platycodon grandiflorum (Jacq.) A. DC. is a famous medicinal plant commonly used in East Asia. Triterpene saponins isolated from P. grandiflorum are the main biologically active compounds, among which polygalacin D (PGD) has been reported to be an anti-tumor agent. However, its anti-tumor mechanism against hepatocellular carcinoma is unknown. This study aimed to explore the inhibitory effect of PGD in hepatocellular carcinoma cells and related mechanisms of action. We found that PGD exerted significant inhibitory effect on hepatocellular carcinoma cells through apoptosis and autophagy. Analysis of the expression of apoptosis-related proteins and autophagy-related proteins revealed that this phenomenon was attributed to the mitochondrial apoptosis and mitophagy pathways. Subsequently, using specific inhibitors, we found that apoptosis and autophagy had mutually reinforcing effects. In addition, further analysis of autophagy showed that PGD induced mitophagy by increasing BCL2 interacting protein 3 like (BNIP3L) levels.In vivo experiments demonstrated that PGD significantly inhibited tumor growth and increased the levels of apoptosis and autophagy in tumors. Overall, our findings showed that PGD induced cell death of hepatocellular carcinoma cells primarily through mitochondrial apoptosis and mitophagy pathways. Therefore, PGD can be used as an apoptosis and autophagy agonist in the research and development of antitumor agents.

Transcriptome analysis of Platycodon grandiflorum at different growth years and discovery of key genes for platycodin biosynthesis / 中国中药杂志

Platycodon grandiflorum is a medicinal and edible medicinal material. Our study is aimed to explore the differences in the gene expression of P. grandiflorum in different growth years, and the expression rules of key genes in the biosynthesis of the main active substances of P. grandiflorum. Illumina Hiseq 4000 sequencing platform was used to sequence the transcriptome of P. grandiflorum in different years. Then, 59 654 unigenes were obtained through filtering, assembly, splicing and bioinformatics analysis of the sequencing data, of which 1 671 unigenes were differentially expressed between at least two samples. The results of cluster analysis showed that there was a great difference in the gene expression of P. grandiflorum from one-year-old to two/three-year-old. There were 1 128 different genes between one-and three-year old P. grandiflorum, and only 57 different genes between two-and three-year-old P. grandiflorum. KEGG enrichment results showed that the differential genes of P. grandiflorum in different years were mainly concentra-ted in the biosynthesis of sesquiterpenes and triterpenes, and the biosynthesis of terpenoid skeletons. In the triterpenoid biosynthesis-related pathways, a total of 15 unigenes were identified, involving 5 enzymes. The expression levels of ACAT, HMGR, FDFT1, SQLE decreased with the increase of the growth year of P. grandiflorum. The expression of HMGS was the highest in the one-year-old P. grandiflorum, followed by the three-year-old sample. This study provides useful data for the development of P. grandiflorum, and also provides a basis for the study of related genes in the biosynthetic pathway of platycodin.

Study on Quality Standard of Compound Platycodon grandiflorum Antitussive Tablets / 中国药房

OBJECTIVE： To establish the quality standard of Compound Platycodon grandiflorum antitussive tablets. METHODS： TLC was used to identify the P. grandiflorum， Polygala tenuifolia and Glycyrrhiza uralensis qualitatively in Compound P. grandiflorum antitussive tablets. HPLC-ELSD method was used to measure the content of platycodin D in Compound P. grandiflorum antitussive tablets. The determination was performed on Agilent C18 column with mobile phase consisted of acetonitrile-water （26 ∶ 74， V/V） at the flow rate of 1.0 mL/min. ELSD was used with drift tube temperature of 105 ℃， gas flow rate of 3.0 L/min and column temperature at 35 ℃. RESULTS： TLC chromatograms of P. grandiflorum， P. tenuifolia and G. uralensis had clear spots with good separation and no same spot from negative samples. The linear range of platycodin D was 0.421 9- 5.062 8 μg （r＝0.999 9）. The quantitative limit and detection limit were 0.364， 0.109 μg， respectively. RSDs of precision， stability， reproducibility and durability tests were all lower than 3.0％. The recovery rates were 87.32％-91.96％ （RSD＝1.73％，n＝6）. The platycodin D contents of 178 samples ranged from 0.004 to 0.73 mg/tablet. The content of platycodin D in 55 batches （30.9％） of samples was lower than the content limit （0.10 mg/tablet） proposed in this study. CONCLUSIONS： Established method is accurate and reliable， and can be used for the quality control of Compound P. grandiflorum antitussive tablets.

Mitigation effects of red Platycodon grandiflorum extract on lipopolysaccharide-induced inflammation in splenocytes isolated from mice / 한국영양학회지

PURPOSE: Platycodon grandiflorum (PG) is known to have effective antimicrobial and anticancer activity. The main bioactive components of PG are saponins, and these could contribute to anti-inflammatory activity. However, little is known about the anti-inflammatory effect of PG. In this study, we aim to assess the anti-inflammatory response to Red PG Extract (RPGE) in splenocytes under ex vivo conditions. METHODS: The cell viability of isolated splenocytes taken from mice was analyzed by performing a Cell Counting Kit-8 assay. The productions of nitric oxide (NO) and cytokines (specifically interleukin-6 (IL-6) and interleukin-10 (IL-10)) were measured utilizing Griess reagent and ELISA, respectively. RESULTS: We found that co-treatment with RPGE and Lipopolysaccharide (LPS) decreased isolated splenocyte proliferation as compared with that of the LPS-stimulated control. We also observed that RPGE markedly suppressed NO synthesis and IL-6 production that was induced by LPS. There were no significant differences of IL-10 production between co-treatment with RPGE plus LPS and treatment with LPS alone. CONCLUSION: When taken together, our data has shown that RPGE mitigates LPS-induced inflammation in splenocytes isolated from mice. Further research is surely needed to confirm the anti-inflammation effects of RPGE in an in vivo model.

Effects of Platycodon grandiflorum/pepper intercropping on root growth, yield and quality of Platycodon grandiflorum / 中国中药杂志

In order to investigate effects of Platycodon grandiflorum and pepper intercropping on root growth, yield and quality of P. grandiflorum, field experiments were conducted in the soils of continuously cultivated P. grandiflorum for three years. The cultivation model was designed as monoculture and intercropping. The monoculture of P. grandiflorum was denoted as CK and the intercrop association of P. grandiflorum/pepper was arranged as follow: in intercrops every two rows of pepper was planted between every three, four and five rows of P. grandiflorum, respectively, and denoted as JC₃₂, JC₄₂ and JC₅₂. Results showed that taproot length and diameter of P. grandiflorum in intercropping association of JC₃₂ was higher than those of P. grandiflorum in monoculture association. This fact suggested that P. grandiflorum intercropped with pepper facilitated its root growth. Compared with monoculture association, the number of lateral root in intercropping association was significantly decreased and the location of lateral root at taproot also altered. This fact suggested that P. grandiflorum intercropped with pepper enhanced appearance quality of P. grandiflorum root. Total root yield and taproot yield of P. grandiflorum in JC₄₂ and JC₅₂ intercropping associations were increased by 4.88%, 8.91% and 14.23%, 12.92%, respectively, compared with monoculture, while root rot incidence decreased significantly. Compared with JC₅₂ intercropping association, JC₄₂ intercropping association significantly increased total saponin and protein contents of P. grandiflorum, decreased root rot incidence, but did not affect taproot yield significantly. Considering root yield and quality, when P. grandiflorum planted in the soil having continuously cultivated P. grandiflorum for three years, the optimal cultivation model was every two rows of pepper was planted between four rows P. grandiflorum.

Inhibitory effect of the aqueous extract of a tetraploid ‘etteum’ variety in Platycodon grandiflorum on degranulation and inflammatory mediator release in RBL-2H3 cells / 한국영양학회지

PURPOSE: Platycodon grandiflorum (a domestic diploid variety, DV-PG) has been used as a food and component of various traditional oriental medicines. Although DV-PG is known to have an anti-allergic effect, little is known about the beneficial health effects of the tetraploid ‘Etteum’ variety in the Platycodon grandiflorum (TV-PG), which is a recently developed variety. In this study, we investigated the effect of TV-PG on the rat basophilic leukemia mast cell (RBL-2H3)-mediated allergic response. METHODS: To examine the effects of TV-PG on the allergic response, RBL-2H3 cells were sensitized with dinitropheny (DNP)-immunoglobin E, treated with various concentrations of TV-PG, and challenged with DNP-human serum albumin. We estimated cell granulation by measuring the release of β-hexosaminidase and production of inflammatory mediators by ELISA. RESULTS: TV-PG had no effect on the proliferation or cytotoxicity of RBL-2H3 cells within the concentration range of 0 to 200 µg/mL. TV-PG inhibited degranulation of RBL-2H3 cells by antigen stimulation in a dose-dependent manner. TV-PG also suppressed the production of inflammatory cytokines and mediators such as interleukin-4, tumor necrosis factor-α, prostagladin E2, and leukotriene B4 in RBL-2H3 cells by antigen stimulation. CONCLUSION: These results indicate that TV-PG exhibits anti-allergic activity via inhibition of degranulation as well as suppression of inflammatory mediators and cytokine release. These findings suggest that TV-PG may have potential as a preventive and therapeutic agent for the treatment of various allergic diseases.

Comparison of the Content of Total Saponins and Platycodin D in Platycodon grandiflorum from Sichuan of Different Cultivated Years / 中国药房

OBJECTIVE：To establish a method for content determination of total saponins and platycodin D in Platycodon grandiflorum from Sichuan and investigate the difference of 2 indexes in P. grandiflorum from Sichuan of different cultivated years. METHODS：The content of total saponins in P. grandiflorum from Sichuan was determined by UV spectrophotometry. The content of platycodin D was determined by HPLC. The contents of total saponins and platycodin D were compared among each 10 samples of annual，biennial and triennial P. grandiflorum from Sichuan. RESULTS：Average contents of total saponins in annual，biennial and triennial P. grandiflorum from Sichuan were 2.47％ ，3.01％ ，2.47％ ，respectively；average contents of platycodin D were 0.23％，0.27％，0.33％，respectively. The contents of 2 indexes in annual P. grandiflorum from Sichuan were in relative low level， while the content total saponins in biennial P. grandiflorum from Sichuan was the highest；the content of platycodin D in triennial P. grandiflorum was the highest. CONCLUSIONS：The contents of indexes are different among P. grandiflorum from Sichuan of different cultivated years. But there is no correlation between them. It is suggested to select biennial and triennial P. grandiflorum from Sichuan.

Protective effects of Platycodon grandiflorum total saponins on acute lung injury in rats and related mechanisms / 中华急诊医学杂志

Objective To investigate the protective effects of Platycodon grandiflorum total saponins (PGTS) on acute lung injury (ALI) in rats and the related mechanisms.Methods Total of 60 SD rats were randomly (random number) divided into control group,model group,low-,middle-and high-dose PGTS group,and dexamethasone group,10 rats in each group.The latter 4 groups and dexamethasone group were injected with 50,100,200 mg/kg PGTS and 5 mg/kg dexamethasone,respectively.After 1 h,the latter 5 groups were intraperitoneally injected with mg/kg LPS to establish the ALI model.The clinical symptoms of the rats were observed.After 12 h,the arterial PaO2 and PaCO2,serum TNF-α and IL-10 level,lung wet/dry weight ratio (W/D),lung tissue SOD,GSH-Px and MDA level and NF-κB protein expression were determined.Results Rats in model group manifested noticeable symptoms of acute lung injury (ALI) and lung tissue lesions.In treatment group with appropriate PGTS dose,ALI symptoms and lung lesions were significantly alleviated,arterial PaO2 was markedly increased (P ＜ 0.05),PaCO2 was decreased obviously (P ＜ 0.05),serum TNF-α level was prominently decreased (P ＜ 0.05),IL-10 level was strikingly decreased (P ＜ 0.05),lung W/D ratio was significantly decreased (P ＜0.05),lung tissue SOD and GSH-Px level were distincdy increased (P ＜0.05),MDA was clearly decreased (P ＜ 0.05),and NF-κB protein expression was plainly decreased (P ＜ 0.05),compared with model group.Conclusions PGTS has undoubted protective effects on acute lung injury induced by LPS in rats.The mechanism may be associated with its role of anti-inflammation,anti-lipid peroxidation and down regulation of NF-κB protein level in lung tissue.

Evolution and transition of quality evaluation of Platycodon grandiflorum / 中国中药杂志

Platycodon grandiflorum, a traditional Chinese medicine commonly used for coughing and eliminating phlegm to relieve asthma, has gained great attention to its quality evaluation in ancient and modern herbal books. This paper investigated the methods of quality evaluation in the ancient herbal books systematically, and the results showed that there were bitter and sweet P. grandiflorum, south and north P. grandiflorum. Those distributed in east China were called south P. grandiflorum, with bitter taste; and those distributed in north China, northeast China were called north P. grandiflorum, with sweet taste. There was a common sense that P. grandiflorum with a bitter taste had good quality in the ancient herbal books, namely those produced in east China. P. grandiflorum of better quality were characterized by thicker and longer root, white color, solid texture and bitter taste in properties. In addition, the quality of P. grandiflorum was also affected by its germplasm, collecting and processing. This paper summarized the formation and development of the "assessing the quality by distinguishing features of traditional Chinese medicinal materials" views of P. grandiflorum, providing the herbalism basis for its present quality evaluation.

Determination of platycodin in Platycodi Radix from different habitats / 中草药

Objective: To establish a method of the determination of total platycodin and platycodin D; And to determine the contents of total platycodin and platycodin D in Platycodi RadiX from different habitats. Methods: The content of total platycodin was determined by 1ultrasonic extraction and weight method; And the content of platycodin D was determined by HPLC. The chromatographic conditions were Waters Symmetry-C18 column, mobile phrase of CH3CN-H2O (22:78), flow rate of 1.0 mL/min, detection wavelength at 210 nm and column temperature 30℃. Results: There was a greater difference between the contents of total platycodin and platycodin D in Platycodi Radix from different habitats. In measuring Platycodi Radix, the content of total platycodin in Platycodi Radix from Zhejiang Province was the highest, which was 12.03%; the contents in Platycodi Radix from Heilongjiang, Liaoning, Anhui, Shandong, and Jiangxi Provinces were more than 6%; However, the contents in Platycodi Radix from the other provinces were lower than 6%, and it was the lowest in Platycodi Radix from Henan province, which was 1.57%. The contents of platycodin D in Platycodi Radix from Yunnan province was the highest, and it was the lowest in Platycodi Radix from Hebei province. Conclusion: The methods could be used for the determination of total platycodin and platycodin D in Platycodi Radix because of theirs simple operations with accurate and reliable results. The contents of total platycodin and platycodin D are different obviously in Platycodi Radix from different habitats, and the contents of total platycodin and platycodin D in the same drug with the level of inconsistency don't show correlation.

Optimized Extraction of Total Saponins from Platycodon grandiflorum / 医药导报

Objective To optimize the process of extracting total saponins from Platycodon grandiflorum by orthogonal experimental design and central composite design-response surface method. Methods Independent variables were concentration of ethanol, reflux extraction time, reflux time and solvent fold. Dependent variables were extraction rate of total saponins in Platycodon grandiflorum and the yield of dry extract.Linear or nonlinear mathematic models were used to estimate the relationship between independent and dependent variables. Results The result of the orthogonal experimental design was 70%ethanol, 180 min for reflux, 6 fold of solvent and 2 times for extraction. The result of central composite design-response surface method was 70% ethanol, 140 min for reflux, 10 fold of solvent and 2 times for extraction. Conclusion The central composite design-response surface method is better than orthogonal experimental design in studying extraction process of Platycodon grandiflorum, which provides basis for feasibility of application of central composite design-response surface method in optimization of extraction process of Chinese traditional medicine.

Effects of different soaking treatments on seed germination and seedling growth of Platycodon grandiflorum / 中草药

Objective: To investigate the effects of different soaking treatments by KNO3, KMnO4, H2O2, GA3, and distilled water for different times on seed germination and seedling growth of Platycodon grandiflorum. Methods: Adopting a double-layer filter paper culture method, the seeds were cultured in the 12 h illumination light incubator at 25 °C, and the germination energy, germination percentage, germination index of the seeds, and the root length and the shoot height of the seedling were recorded. Then the data were analyzed. Results: The best soaking treatment to break seed dormancy, promote the seed germination, and improve the seedling growth of P. grandiflorum was with 0.150 g/L GA3 for 24 h. In addition, another effective soaking treatment is using 0.005 g/mL KNO3 for 12 h. Conclusion: The appropriate soaking reagent and time for the seed germination of P. grandiflorum are obtained, which could provide the guidance for seedling and artificial cultivating P. grandiflorum.

HPLC fingerprint of Platycodon grandiflorum in different picking times / 中成药

AIM:To establish HPLC fingerprints of Platycodon grandiflorum in different picking times in Henan province.METHODS:HPLC chromatography condition:Hypersil C_(18) column(250 mm×4.6 mm,5 μm);the mobile phase was acetonotrile with 0.05% phosphoric acid and the gradient elution mode was applied in chromatographic separation;The flow rate was 0.5 mL/min;The temperature of column was 30℃;the detective wavelength was set at 210 nm;RESULTS:The overall similarity in samples collected in autumn was higher than that in spring,and autumn was the appropriate for harvest.CONCLUSION:This method is simple and accurate with a good reproducibility.It provids a reliable scientific basis for the quality control of Platycodon grandiflorum.

Growth Inhibitory Effects of Various Herbal Extracts on Metronidazole Resistant Strain of Trichomonas vaginalis / 감염과화학요법

BACKGROUND: Metronidazole has been known as the most effective drug for treatment of Trichomonas vaginalis-related diseases. However, it has been reported that metronidazole has adverse effects and incidence of metronidazole-resistant T. vaginalis (CDC085) has increased. Development of new drug, which is effective against metronidazole-resistant T. vaginalis and showing no adverse effects, has been required. METHODS: The purpose of this study was to investigate effects of various extracts from herbs such as Quisqualis indica, Gleditsia sinensis, Prunus armeniaca, Morus alba, Platycodon grandiflorum, Ailanthus altissima, Stemona japonica, Biota orientalis, Dryobalanops aromatica, and Cimicifuga heracleifolia on metronidazole resistant strain of T. vaginalis in vitro (CDC085). RESULTS: Anti-Trichomonas activities were observed in T. vaginalis treated with G. sinensis, P. armeniaca, and P. grandiflorum on the growth and fine structure of metronidazole resistant strain of T. vaginalis. Of the three standard extracts that showed the most effective anti-trichomonas activity, G. sinensis was the most effective. The inhibitory effects of fraction extracts of this drug were shown on the growth of T. vaginalis. The fine structure of the cytoplasm was changed after application of G. sinensis extract. The number of polyribosome and hydrogenosome decreased whereas the number of food vacuole and vacuole in the cytoplasm increased, compared with that of untreated control group. CONCLUSION: The results of our study indicate that G. sinensis may induce the inhibition of cell multiplication as well as impairment of protein synthesis of metronidazole resistant strain of T. vaginalis in vitro.

Growth Inhibitory Effects of Various Herbal Extracts on Metronidazole Resistant Strain of Trichomonas vaginalis / 감염과화학요법

BACKGROUND: Metronidazole has been known as the most effective drug for treatment of Trichomonas vaginalis-related diseases. However, it has been reported that metronidazole has adverse effects and incidence of metronidazole-resistant T. vaginalis (CDC085) has increased. Development of new drug, which is effective against metronidazole-resistant T. vaginalis and showing no adverse effects, has been required. METHODS: The purpose of this study was to investigate effects of various extracts from herbs such as Quisqualis indica, Gleditsia sinensis, Prunus armeniaca, Morus alba, Platycodon grandiflorum, Ailanthus altissima, Stemona japonica, Biota orientalis, Dryobalanops aromatica, and Cimicifuga heracleifolia on metronidazole resistant strain of T. vaginalis in vitro (CDC085). RESULTS: Anti-Trichomonas activities were observed in T. vaginalis treated with G. sinensis, P. armeniaca, and P. grandiflorum on the growth and fine structure of metronidazole resistant strain of T. vaginalis. Of the three standard extracts that showed the most effective anti-trichomonas activity, G. sinensis was the most effective. The inhibitory effects of fraction extracts of this drug were shown on the growth of T. vaginalis. The fine structure of the cytoplasm was changed after application of G. sinensis extract. The number of polyribosome and hydrogenosome decreased whereas the number of food vacuole and vacuole in the cytoplasm increased, compared with that of untreated control group. CONCLUSION: The results of our study indicate that G. sinensis may induce the inhibition of cell multiplication as well as impairment of protein synthesis of metronidazole resistant strain of T. vaginalis in vitro.

Comparison of Vasodilator Effects of Platycodin D and D3 in Rats

The aim of the present study was to examine the effects of platycodin D and D3, which are active components derived from the roots of Platycodon grandiflorum A. DC., on the contractile force of the i3olated rat aorta and blood pressure of the anesthetized rat, and also to elucidate its mechanism of action. Both phenylephrine (an adrenergic alpha1-receptor agonist) and high potassium (a membrane- depolarizing agent) caused great contractile responses in the isolated aortic strips. Platycodin D at high concentration (24microgram/ml) inhibited contractile responses induced by phenylephrine (10 (-5) M) and high potassium (5.6x10 (-2) M), while low concentrations of platycodin D (4~8microgram/ml) did not affect those responses. However, platycodin D3 (8~32microgram/ml) did not alter the contractile responses evoked by phenylephrine and high K+. Interestingly, the infusion of platycodin D3 (1.0 mg/kg/30 min) significantly reduced the pressor responses induced by intravenous norepinephrine. However, platycodin D3 (1.0 mg/kg/30 min) did not affect them. Taken together, these results show that intravenously administered platycodin D depresses norepinephrine-induced pressor responses in the anesthetized rat, at least partly through the blockade of adrenergic alpha1-receptors. Platycodin D also caused vascular relaxation in the isolated aortic strips of the rat via the blockade of adrenergic alpha1-receptors, in addition to an unknown direct mechanism. However, platycodin D3 did not affect both norepinephrine-induced pressor responses and the isolated rat aortic contractile responses evoked by phenylephrine and high potassium. Based on these results, there seems to be much difference in the mode of action between platycodin D and platycodin D3.

Effects of Platycodon grandiflorum,Glycyrrhiza uralensis Fisch or the compound of both on the growth of oral pathogens in vitro / 实用口腔医学杂志

Objective: To examine effects of Platycodon grandiflorum (JG), Glycyrrhiza uralensis Fisch (GC) or the compound of JG with GC on the growth of cariogenic and periodontopathic bacteria in vitro . Methods: JG,GC and the compound were extracted with ?=65% ethanol respectively. The minimal inhibitory concentration(MIC)and minimal bactericidal concentration (MBC) of JG, GC or compound against S.mutans MT818, S. sobrinus 6715, P.gingivalis 381 and B.forsythus 43037 were measured by drug sensetivity test.Results: JG showed no effect on the growth of oral pathogens tested;GC inhibited S.mutans MT 8418 and S.sobrinus 6715 with MIC of 3.91 mg/ml.The compound inhibited the microorganisms with the MIC of 1.96 mg/ml against S.mutans MT8148 or S.sobrinus 6715, 3.91 mg/ml against P.gingivalis 381, 7.81 mg/ml against B.forsythus 43037 respectively.Conclusion: The compound of JG and GC has stronger inhibition and bactericidal effects on oral pathogens than the single medicine.

Cryopreservation of in vitro shoot-tips of Platycodon grandiflorum by vitrification and plant regeneration / 中草药

Objective To get a new approach to conserve the germplasm of Platycodon grandiflorum. Methods A method of vitrification was studied to cryopreserve in vitro shoot-tips of P. grandiflorum, and the regenerated plantlets were observed subsequently. Results Shoot-tips, 2—3 mm length, gotten from in vitro shoots of P. grandiflorum precultured on MS medium supplemented with 5% DMSO and 103 g/L sucrose for 3 d, were loaded with the 60% PVS_2 for 20 min at 20 ℃, and incubated in PVS_2 for 90 min at 0 ℃ prior to a direct plunging into liquid nitrogen (LN) and keeping for 1 d. After rapid thawing in water at 40 ℃, the shoot-tips were rinsed in the MS medium supplemented with 410 g/L sucrose for 20 min, and plated on the filter paper sustained by the MS regeneration medium supplemented with 0.6 mg/L KT, 0.2 mg/L BA, and 0.05 mg/L NAA for 1 d in dark and subcultured on the above regeneration medium for one week in dark prior to exposure to the light. The survival of shoot-tips was up to 80%, and they grew normally. Conclusion The method of vitrification to cryopreserve the germplasm of P. grandiflorum is simple in handling with high in survival and normal in regeneration and can be applied in practice.