Screening of effective antioxidant components from Trichosanthes extract and assessment of their antioxidant activity / 南方医科大学学报

OBJECTIVE@#To screen the effective antioxidant components in Trichosanthes extract based on the mean value of Deng's correlation degree and assess the antioxidant activity of the identified components.@*METHOD@#High-performance liquid chromatography (HPLC) was used to obtain the fingerprints of Trichosanthes extract, and the clearance rates of DPPH · and O2-· by 3, 9 and 27 mg/mL Trichosanthes extract were determined. The antioxidant spectrum effect of Trichosanthes extract was analyzed by calculating the mean value of Deng's correlation degree to screen the effective antioxidant component group. According to the contents of each known components in the antioxidant effective component group, mixed solutions of the components were prepared and tested for their clearance rates of DPPH · and O2-·.@*RESULTS@#The 36 common peaks in HPLC fingerprints of Trichosanthes extract showed different degrees of correlation with DPPH · and O2-· clearance. The common peaks with a correlation degree greater than the median value included peaks 21, 36, 8, 31, 14, 5, 27, 2, 24, 15, 18, 33, 22, 34, 35, 19, 28 and 25. The 5 components, namely kaempferol (peak 36), isoquercitrin (peak 8), luteolin (peak 31), rutin (peak 5) and apigenin (peak 35), were tentatively identified to constitute the effective antioxidant component group with a mass ratio 3∶2∶2∶ 1∶1 in Trichosanthes extract. The prepared mixed solutions of antioxidant effective component group (6.12, 2.04, and 0.68 μg/mL) showed clearance rates of DPPH · of 65.4%, 64.0% and 61.0%, and clearance rates of O2-· of 12.9%, 9.5% and 8.3%, respectively.@*CONCLUSION@#We identified the material basis for the antioxidant activity of Trichosanthes and screened the antioxidant effective component group in Trichosanthes extract.

LC-MS/MS analysis of pharmacokinetics of 3, 29-dibenzoyl karounitriol in rats following administration of Trichosanthes sustained-release pellets / 南方医科大学学报

OBJECTIVE@#To study the plasma concentration and pharmacokinetics of 3, 29-Dibenzoyl Karounitriol (3, 29-DK) from sustained- release pellets and extracts of Trichosanthes at different time points in rats using high-performance liquid chromatography- tandem mass spectrometry (LC-MS/MS).@*METHODS@#Healthy male SD rats were given a single gavage of Trichosanthes sustained-release pellets or Trichosanthes extract, and orbital blood samples were taken at different time points within 48 h after drug administration in the pellet group and within 5 h in Trichosanthes extract group for determination of the plasma concentrations of 3, 29-DK using LC-MS/MS. The standard curve of 3, 29-DK content was established, and the specificity, minimum detection limit, precision and accuracy, extraction recovery, stability and matrix effect of LC-MS/MS analysis were assessed. The mean plasms levels of 3, 29-DK at different time points after the drug administration were determined and its pharmacokinetic parameters were calculated using Das 2.0 software.@*RESULTS@#LC-MS/MS analysis showed a good linearity of 3, 29-DK concentration within the range of 0.5-32 ng/mL, and the results of methodological validation confirmed the validity of this method for biological sample determination. Trichosanthes sustained-release pellets and Trichosanthes extract showed significant differences in their AUC, AUC, MRT, MRT, t and T of 3, 29-DK after administration in rats ( < 0.05).@*CONCLUSIONS@#Trichosanthes sustained-release pellets are capable of sustained-release of 3, 29-DK in rats, and thus provides a basis for the study of new dosage forms of Trichosanthes.

Research on rules of syndrome differentiation and medication of coronary heart disease based on data mining / 中草药

Objective: To analyze the rules of coronary heart disease syndrome differentiation based on the data mining technology. Methods: First of all, the famous medical records and prescriptions were collected and normalized to construct the database of coronary heart disease prescriptions. Then, IBM SPSS Statistics 20.0, a statistical software, was used to conduct statistics on the distribution of drug frequency, disease site syndrome frequency and disease syndrome frequency. Finally, the Apriori algorithm was applied to carry out data mining and analyze the underlying law of drug compatibility. Results: A total of 145 prescriptions were selected and analyzed, including 216 Chinese medicines, eight syndromes of disease location and 12 syndromes of disease nature. Forty-six herbs with higher frequency were found, and the top five TCM herbs were Salvia miltiorrhiza, Glycyrrhiza uralensis, Trichosanthes kirilowii, Pinellia ternata and Ligusticum chuanxiong. The main syndromes of disease location were heart, kidney, spleen and liver, while the main syndromes of disease nature were blood stasis, qi deficiency, phlegm, deficiency of yang, qi stagnation, and deficiency of yin. When the minimum support was 15% and the minimum confidence coefficient was 70%, the association rule analysis results showed that a total of 15 association rules for two-herb-combinations and 26 association rules for the trigeminy medications were obtained. The most frequency two-herb-combinations were Allium macrostemon→T. kirilowii", "Schisandra chinensis→Ophiopogon japonicus", "Curcumae Radix→S. miltiorrhiza. The most frequently used trigeminy medications were A. macrostemon + P. ternata→T. kirilowii, S. miltiorrhiza + A. macrostemon→T. kirilowii and A. macrostemon + G. uralensis→T. kirilowii. Conclusion: The syndrome differentiation and medication of TCM in the treatment of coronary heart disease mainly focus on invigorating the circulation of blood and resolving stasis, moving qi, nourishing and replenishing qi, clearing up phlegms. It is consistent with the main syndromes of the disease nature, such as blood stasis and qi deficiency, and the herbs mostly return to the heart meridian, which is consistent with the main syndromes of the disease location. The rules of syndrome differentiation and medication of based on the data mining technique have great value for clinical medication guidance and application to analyze the prescription for coronary heart disease.

Analysis of quality markers based on efficacy and material base of different medicinal parts of Trichosanthes kirilowii / 中草药

Different parts of Trichosanthes kirilowii can all be used as medicines, including the fruits (Trichosanthis Fructus), pericarps (Trichosanthis Pericarpium), seeds (Trichosanthis Semen) and roots (Trichosanthis Radix). Modern research has confirmed that the main active ingredients of Trichosanthis Pericarpium are flavonoids and amino acids; Trichosanthis Semen mainly contains terpenoids and sterols; Trichosanthis Radix mainly contains protein, steroids and polysaccharides. And the pharmacological effects of various medicinal parts are also different. This paper summarizes the traditional efficacy, chemical composition and modern pharmacological effects of different medicinal parts of T. kirilowii, analyzes the relationship between them, so as to analyze and predict the quality marker of T. kirilowii.

Preparation and in vitro release rate of trichosanthes sustained-release pellets / 国际药学研究杂志

Objective: To prepare Trichosanthes sustained-release pellets and investigate its in vitro release. Methods: Trichosanthes sustained-release pellets were prepared by fluid-bed coating technique.The preparation process of its drug loading layer was optimized by orthogonal experiment with binder, solvent, drug loading and creeping speed of peristaltic pump as the factors of investigation and the yield as the investigation index. The optimum prescription and preparation of the pellets were optimized by single factor test with the content of coating material EC, the amount of poreforming agent PEG4000, the amount of talc powder, the coating weight and the curing time as the factors of investigation and the in vitro release as the investigation index. The in vitro release of the pellets was investigated by high performance liquid chromatography with 3, 29-Diphenyl of Trichosanthes Kirilowii Triol(3, 29-DK)as the detection index. Results: The optimum preparation technology of the drug loading layer was 5% binder, 60% ethanol, 35% drug loading and 0.3 r/min peristaltic velocity of fluid-bed peristaltic pump. The optimum preparation technology of sustained-release layer was 5% EC, 1.5% PEG 4000, 1.25% talc powder, 10% weight gain of coating and 6 h curing. Conclusion: The Tricho- santhes sustained-release pellets prepared in this study were released smoothly. Its production method was simple and easy for operation.

Isolation and identification of antitumor constituents from Trichosanthes tricuspidata / 中国药科大学学报

@#Ten compounds were isolated and purified from the dichloromethane fraction of Trichosanthes tricuspidata roots by silica gel and ODS column chromatographies. Their chemical structures were identified on the basis of their physical and chemical properties as well as the spectral data. These isolated compounds were elucidated as khekadaengoside O(1), a new hexanorcucurbitane glycoside, together with nine known compounds, including khekadaengoside C-E, K(2-5), cucurbitacin J-2-O-β-glucopyranoside(6), cucurbitacin K-2-O-β-glucopyranoside(7), cucurbitacin B, J, K(8-10). In addition, the anti-tumor activity of compounds 1-10 were evaluated in hepatocellular carcinoma BEL-7402 cell line. Among them, compounds 8-10 showed potent antitumor activity.

Chemical constitutes from flesh of Trichosanthes kirilowii / 中草药

Objective: To investigate the chemical constituents from the flesh of Trichosanthes kirilowii. Methods: The chemical constituents from n-butanol fraction of 95% ethanol extract of the flesh of T. kirilowii were separated by silica gel and ODS chromatogram columns as well as preparative HPLC. On the basis of NMR and MS data analysis, their structures were elucidated. Results: Twenty-six compounds were isolated from 95% ethanol extract of T. kirilowii, of which six organic esters were ethyl laurate (1), dibutyl phthalate (2), diethyl ethaneioate (3), dibutyl-2-malate (4), 6,10,14,18-tetramethyl-2-ethyl-7-ene-3-hydroxyl-ninecanol- 1-butyl ester (5), drechslerol-B (6), nine organic acids and phenolic acids were p-hydroxybenzaldehyde (7), salicylic acid (8), vanillic acid (9), isovanillic acid (10), protocatechuate (11), trans-cinnamic acid (12), p-hydroxycinnamic acid (13), trans-ferulic acid (14), and lauric acid (15), eight flavonoids were diosmetin (16), apigenin (17), chrysoeriol (18), luteolin (19), 4’-hydroxyscutellarin (20), quercetin (21), diosmetin-7-O-β-D-glucoside (22), chrysoeriol-7-O-β-D-glucoside (23), two aldehydes were 5-acetoxymethyl- 2-furaldehyde (24), 5-hydroxymethyl furfural (25) and one cycloaltinol compounds was cyclotucanol 3-palmitate (26). Conclusion: All compounds except compound 10 are isolated from the Trichosanthes genus for the first time.

Screening and validation of antithrombotic effective components group from Trichosanthes extract / 药学学报

To screen the antithrombotic effective components group of Trichosanthes extract, and to verify its pharmacodynamics and analyze its mechanism, the HPLC fingerprint of Trichosanthes extract (0.09, 0.45, 0.9 g·kg-1) was established, and the pharmacodynamic indexes of antithrombosis in rats with aspirin (0.01 g·kg-1) as positive control group were determined (the animals used in this experiment were approved by the Medical Ethics Committee of Wannan Medical College). The antithrombotic spectrum-activity relationship of Trichosanthes extract was studied and the effective antithrombotic ingredients group was screened by grey relational analysis. The monomer compound mixed solution (0.006, 0.03, 0.06 g·kg-1) was prepared according to the content of each component in the active component group, and the pharmacodynamics and action mechanism were studied to verify the correctness of the spectrum-effect relationship. The correlation between the 22 components of Trichosanthes extract and antithrombotic efficacy was different and showed dose-effect relationship. Cytosine, uracil, guanine, hypoxanthine, xanthine, adenine, guanosine, and adenosine are the main antithrombotic components of Trichosanthes extract. The ratio of cytosine, uracil, guanine, hypoxanthine, xanthine, adenine, guanosine and adenosine was 3∶12∶10∶5∶2∶8∶13∶14. Compared with the model group, the thrombus dry weight of each effective components group could be effectively reduced (P<0.01 or P<0.05), but there was no significant difference between each effective components group and the Trichosanthes extract group. Compared with the model group, the TXB2 content in group (0.06 g·kg-1, 0.03 g·kg-1) could be effectively reduced (P<0.01 or P<0.05), and the content of 6-keto-PGF1α could be increased in each group (P<0.01), and the TXB2/6-keto-PGF1α tended to be normal and showed a dose-effect relationship. The effect was better than that in the Trichosanthes extract group (0.45 g·kg-1) (P<0.01). The effective ingredients group has a good antithrombotic effect, its mechanism is to inhibit platelet aggregation and improve vascular endothelial function.

Predicting and validating the mechanism of Trichosanthes mediated anti-myocardial ischemia-reperfusion injury by network pharmacology / 药学学报

Network pharmacology and rat ischemia-reperfusion injury (MIRI) model was used to analyze the mechanism of cardiac protection by Trichosanthes. The animal experiments were approved by the Medical Ethics Committee of Wannan Medical College. Compounds were screened by TCMSP database and TCM Database@ Taiwan according to oral bioavailability (OB > 30%) and drug like activity (DL > 0.18). The PDBID value of the compound (Z'-score < 0.5) was obtained in DRAR-CPI database and converted into a target protein by UniProt database. Human genes of target proteins were identified using the term " myocardial ischemia reperfusion injury" as the keyword through the CoolGeN database. GOTERM_BP _ DIRECT enrichment analysis of target proteins related to MIRI and KEGG PATHWAY annotation analysis were performed using the DAVID database. The component-target protein-signal pathway network was constructed using Giphi0.9.2 software. The expression of mitogen-activated protein kinase (MAPK) signaling pathway-related proteins in MIRI rats pretreated with Trichosanthes (0.2, 1.0 and 2.0 g·kg-1) was analyzed by Western blot with compound Danshen (85.05 mg·kg-1) as a positive control. Network pharmacology found that 12 compounds, including schottenol in Trichosanthes, synergistically inhibit MIRI through multiple targets or biological pathways, involving target proteins such as extracellular regulated protein kinase 2 (ERK2), c-jun-N-terminal kinase-1 (JNK1) and p38MAPK in MAPK signaling pathways. Western blot results showed that phosphorylation of ERK1/2 was dose-dependently up-regulated in MIRI rats pretreated with Trichosanthes, while the level of p38MAPK or JNK1 phosphorylation was down-regulated in a dose-dependent manner. Compared with the control group, phosphorylation of ERK1/2, JNK1 and p38MAPK protein showed significant difference in medium and high dose groups (1.0 and 2.0 g·kg-1) (P<0.01). Therefore, Trichosanthes could play an anti-MIRI role by regulating phosphorylation of ERK1/2, JNK1 and p38MAPK proteins in rats. In conclusion, the targets and pathways of Trichosanthes on anti-MIRI were revealed by network pharmacology and verified in rat MIRI model, providing the scientific basis for further study on the mechanism of Trichosanthes for cardiac protection.

Evaluation of root-knot nematode disease control and plant growth promotion potential of biofertilizer Ning shield on Trichosanthes kirilowii in the field

Abstract Biofertilizer Ning shield was composed of different strains of plant growth promotion bacteria. In this study, the plant growth promotion and root-knot nematode disease control potential on Trichosanthes kirilowii in the field were evaluated. The application of Ning shield significantly reduced the diseases severity caused by Meloidogyne incognita, the biocontrol efficacy could reached up to 51.08%. Ning shield could also promote the growth of T. kirilowii in the field by increasing seedling emergence, height and the root weight. The results showed that the Ning shield could enhance the production yield up to 36.26%. Ning shield could also promote the plant growth by increasing the contents of available nitrogen, phosphorus, potassium and organic matter, and increasing the contents of leaf chlorophyll and carotenoid pigment. Moreover, Ning shield could efficiently enhance the medicinal compositions of Trichosanthes, referring to the polysaccharides and trichosanthin. Therefore, Ning shield is a promising biofertilizer, which can offer beneficial effects to T. kirilowii growers, including the plant growth promotion, the biological control of root-knot disease and enhancement of the yield and the medicinal quality.

Genetic diversity of Trichosanthis Fructus for seeds cultivating / 中草药

Objective To reveal the genetic diversity of cultivating resources of Trichosanthes sp. from different areas. Methods The genetic background of 11 populations (including 30 samples) of Trichosanthis Fructus were researched by both ITS and SRAP molecular markers. The software mega 5.0 and NTSY-pc 2.1 were taken to analysis the ITS sequences and SRAP polymorphic bands data respectively. Results There are obvious mutation in ITS sequences among T. kirilowii and T. rosthornii, ITS sequences could authenticate the different resources in some extent. 165 SRAP bands were amplified by using the optimized 15 primer pairs, and 136 were polymorphic bands, the polymorphic rate was up to 82.4%. The samples were separated into two clusters at the genetic similarity coefficient of 0.18, one cluster gathered the samples of Yuelou Num.8 (T. kirilowii) and the other cluster further separated into three sub-clusters at the coefficient of 0.90, which were mainly T. rosthornii. Conclusion There are several species of Trichosanthes have been used to harvest the seeds in China, and the major one is T. rosthornii. The cultivating resources of T. rosthornii have little difference in genetic backgrounds.

Based on infrared spectrum to study the changes in the intrinsic chemicalcomponents in Trichosanthes before and after steaming / 国际药学研究杂志

Objective To analyze the change in infrared(IR)spectral information and to screen out the classification of major compounds affecting information difference in IR spectra of Trichosanthes before and after steaming. Methods The similarity between the original IR fingerprint and the first derivative IR fingerprint of Trichosanthes and their steamed products was calculated by using the Computer Aided Similarity Evaluation System. The principal component analysis(PCA)model and the partial least squares dis-criminant analysis(PLS-DA)model of IR spectral data of Trichosanthes before and after steaming were established by using SIMCA-P 11 statistical software for PCA and PLS-DA,and the classification of major compounds affecting information difference in IR spectra of Trichosanthes before and after steaming was selected by 3D scatter plot,load 3D scatter plot and variable important in project(VIP) value. Results The similarity between the original IR fingerprint and first derivative IR fingerprint of Trichosanthes and their steamed products were 0.9165 and 0.2832. Seven VIP>1 spectral peaks were screened out by using SIMCA-P 11 statistical software,of which,the absorption peak of 1456 cm-1 was νc=c,the absorption peak of 1726 cm-1 was νc=o and the VIP values were 1.6290 and 1.4256 respectively. Conclusion The categories of compounds of Trichosanthes before and after steaming did not change,but the chemical components changed. Compounds of Trichosanthes before and after steaming affect the difference in IR spectral information may mainly contain C=C-C=C or C=O or both of them.

Analysis of the changes in the chemical components in Fructus Trichosanthes, before and after the processing of steaming,based on the PCA and PLS-DA algorithms / 国际药学研究杂志

Objective To analyze the changes in the chemical components in Fructus Trichosanthes before and after the pro?cessing of steaming,so as to explore the material basis of the pharmacodynamic changes between Trichosanthes and steamed Tricho-santhes.Methods The peaks matching data of Fructus Trichosanthes and steamed Fructus Trichosanthes were obtained by using the similarity evaluation system for chromatographic fingerprints of traditional Chinese materia medica.The principal component analysis (PCA)model and the partial least squares discriminant analysis(PLS-DA)model for the analysis of the Fructus Trichosanthes and steamed Fructus Trichosanthes data were established using the SIMCA-P 11 statistical software for PCA and PLS-DA,from which the score chart,load chart and Variable Importance(VIP)value were obtained,so as to identify the main different components in Fructus Trichosanthes and steamed Fructus Trichosanthes.Results The PCA(R2X=0.96,Q2=0.552)model and PLS-DA(R2Y=0.917,Q2=0.579)model were established,and 8 chromatographic peaks with significant difference in peak area were selected.Among them,two of the chromatographic peaks were assigned to be 5-hydroxy methyl furfural and vanilla acid,and 5-hydroxy methyl furfural had the largest VIP value.In addition,an unknown component was also found in the steamed Fructus Trichosanthes,which was generated in the process of steaming and needed to be identified in future studies.Conclusion The content of some chemical components in Fruc?tus Trichosanthes were changed after the process of steaming,and the processing of steaming also caused the formation of an unknown chemical component.5-Hydroxy methyl furfural and vanillic acid seem to be a likely choice for exploring the material basis of the phar?macodynamic changes in Fructus Trichosanthes after the processing of steaming.

Comparison of Contents of Adenine and Guanosine in Peels of Three Strains of Trichosanthes kirilowii Maxim / 中国中医药信息杂志

Objective To compare the contents of guanosine and adenine in peels of three strains of Trichosanthes kirilowii Maxim.;To provide references for germplasm resources evaluation and breeding of the Chinese medicinal materials. Methods HPLC method was used to determine the contents of guanosine and adenine with Waters AtlantisT3-C18 column (4.6 mm×150.0 mm, 5 μm). The mobile phase was methanol and water, and the detection wavelength was 254 nm. Results The contents of guanosinein in peels of three strains of Trichosanthes kirilowii Maxim. were 0.159 4, 0.159 6, 0.134 1 mg/g, respectively;the contents of adenine were 0.097 1, 0.127 9, 0.093 4 mg/g, respectively. Conclusion There were significient differences in the contents of adenine among three strains of Trichosanthes kirilowii Maxim.. Strain Ⅱ is with higher level of guanosine and adenine, which implies that the breeding of the Chinese medicinal materals is one way to improve the quality of Trichosanthis Exocarpium.

Separation of flavonoids from stems and leaves of male plants in Trichosanthes kirilowii and study on their activity of scavenging DPPH free radical / 中草药

Objective: To study the flavonoids of the stems and leaves of male Trichosanthes kirilowii and to make a primary research on the structure activity relationship between flavonoids and their DPPH-scavenging capacity. Methods: Flavonoids from the stems and leaves of male T. kirilowii were separated by chromatographic techniques, such as polyamide resin, high-speed countercurrent chromatography and high performance liquid chromatography. According to the chemical properties and spectral analysis, the chemical structures of the compounds were identified. And we determined the antioxidant ability in vitro of seven flavonoids by DPPH methods. Results: Seven flavonoids were isolated from the stems and leaves of male T. kirilowii. They were luteolin (1), chrysoeriol (2), luteolin-7-O-β-D-glucoside (3), chrysoeriol-7-O-β-D-glucoside (4), apigenin-7-O-β-D-glucoside (5), diosmetin-7-O-β-D-glucoside (6), and quercetin-3-O-β-glucoside (7). Under the present experimental condition, the order of their DPPH-scavenging capacities was 1 > 3 > 7 > 2 > 4> 6 > 5. Conclusion: Compounds 1, 2, 6, and 7 are isolated from this part of male T. kirilowii for the first time. DPPH-scavenging capacities of the compounds 1, 3, and 7 are much stronger than others, but they all have 3',4' two adjacent hydroxide groups in B ring on the view of structure. DPPH-scavenging capacities of compounds 4 and 6 are much weaker than compound 3, but the former have 3' or 4' hydroxyl methylation in the structure. DPPH-scavenging capacity will also decrease if there is a 7 hydroxyl glycosylation in ring A by comparing compound 1 and 3. We speculate that it is caused by the increase of the steric hindrance.

Comprehensive quality evaluation of medicinal parts of Trichosanthes kirilowii from different populations / 中草药

Objective: To analyze the quality of medicinal parts of Trichosanthes kirilowii from different populations and to establish a new method to evaluate the medicinal material quality. Methods: Contents of protein, flavonoids, and polysaccharide were analyzed by AA3 Continuous Flow, ultraviolet-visible spectrophotometry, and sulfuric acid-phenol. The 3, 29-dibenzoyl rarounitriol (3, 29-DR) content in Trichosanthis Semen and the cucurbitacin bcontent in Trichosanthis Radix were determined by RP-HPLC. In addition, the quality of medicinal materials was evaluated by the principal components analysis (PCA) and cluster analysis. Results: The quality of Trichosanthis Semen was the best in T. kirilowii from Henan Anyang-Liyuan with highest contents of 3, 29-DR and protein; Shanxi Jiang County T. kirilowii can be better used as Trichosanthis Pericarpium with higher contents of protein and polysaccharide; T. kirilowii from Anhui Yuexi-Heidapian could be regarded as Trichosanthis Radix for cultivating, because of higher protein content, lower starch content, and medium cucurbitacin bcontent. Conclusion: The PCA and cluster analysis are effective in evaluating the medicinal material quality. The newly established model will bring the significant benefits for evaluating the quality of T. kirilowii.

Cloning and sequence analysis of squalene synthase gene from Trichosanthes rubriflos / 中草药

Objective: To clone the full length cDNA encoding squalene synthase (SS) from Trichosanthes rubriflos and to carry on its sequence analysis, so as to lay the foundation for the further study on the positively selected sites and function correlation analysis of SS which is the key enzyme for triterpene synthesis pathway. Methods: According to the cDNA comparison on SS gene from Gynostemma pentaphyllum and Siraitia grosvenorii, 5'-upstream degenerate primers of the cDNA of SS gene from T. rubriflos were designed and the full length cDNA of SS gene from T. rubriflos was amplified by 3'RACE kit. Results: The full length cDNA of SS gene from T. rubriflos composed of 1 466 nucleotides was obtained. The open reading frame (ORF) of SS gene from T. rubriflos was 1 254 bp in length, corresponding to a predicted polypeptide of 417 amino acid residues. The results of homologous alignment analysis in GenBank demonstrated that the cDNA sequence of SS gene from T. rubriflos had 78%-94% similarity on the nucleotide sequence compared with SS from known plant and 74%-93% similarity on the deduced amino acid sequence compared with SS gene from other plants. Conclusion: The full length cDNA of SS gene from T. rubriflos has been cloned, which not only lays a foundation for the further study on the gene expression, gene structure, and gene mutation, but also provides the important data base for the association study between the positively selected sites and function correlation analysis of which is the key enzyme for triterpene synthesis pathway.

Effects of extractive pericarpium trichosanthes on apoptosis of HUVECs induced by high glucose / 中国药理学通报

Aim To study the inhibitory effects of ex-tractive pericarpium trichosanthes ( EPT) on high glu-cose-induced apoptosis in human umbilical vein endo-thelial cells ( HUVECs ) and its underlining mecha-nisms. Methods HUVECs were cultured. Effects of EPT at different concentrations on the high-glucose-in-duced apoptosis in HUVECs were observed. The cell viability of HUVECs was determined by MTT colori-metric method. Cell apoptosis was identified by Ho-echst staining. The intracellular activity of Caspase-3 was detected with colorimetry. Protein expression and p65 nuclear translocation of NF-κB were detected by Western blot and immunofluorescence staining. Re-sults Treated with 30 mmol · L-1 glucose for 48 hours had a significantly decrease on cell viability com-pared with control, and the apoptotic rate and Caspase-3 activity were increased markedly, the protein expres-sion of NF-κB was upregulated and p65 nuclear trans-location in HUVECs increased; Pre-incubation with EPT(12. 5,25,50 mg·L-1 ) for 1 hour enhanced the cell viability, and decreased the apoptotic rate and Caspase-3 activity and downregulated the expression of NF-κB in a dose-dependent manner. Moreover, EPT could inhibit p65 translocation. Conclusion EPT can protect HUVECs against the apoptosis induced by high glucose in vitro,and its mechanism may be related with downregulation of NF-κB expression and inhibition of the intracellular activity of Caspase-3 .

Content Comparison of Amino Acid in Female and Male Plants of Trichosanthes Kirilowii / 中国药师

Objective:To compare the content of amino acid in female and male plants of Trichosanthes kirilowii. Methods:Total protein was hydrolyzed by hydrochloric acid, and then determined by HPLC. Results:The percentage of amino acid in female and male plants of Trichosanthes kirilowii was 7. 35% and 7. 04%, respectively. Conclusion:The method provides a new idea for the determi-nation of amino acid in the root of Trichosanthes kirilowii. The total amino acid content in the root of male plants is lower than that of fe-male plants, while the content of alkaline amino acid is opposite. The content of histidine in male plants is about twice as much as that in female plants. The essential amino acid content has no significant difference.

Chemical constituent changes of Aconiti Cocta Radix before and after combination with Trichosanthes Fructus in different ratios by RRLC-QTOF/MS / 中草药

Objective: To analyze the chemical constituent changes of Aconiti Cocta Radix (ACR) before and after combination with Trichosanthes Fructus (TF) in different ratios and to provide the material bases for the incombination. Methods: The rapid resolution liquid chromatography with quadrupole and time-of-flight mass spectrometry (RRLC-QTOF/MS) was used to analyze the extract solution from ACR and TF combination. The chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C18 column (100 mm × 2.1 mm, 1.8 μm) with 0.1% formic acid water solution-0.1% formic acid in acetonitrile solution as mobile phase for gradient elution. Mass Hunter Workstation and partial least squares-discriminant analysis (PLS-DA) were utilized for the identification of components and chemical biomarkers. Peak-area ratio of the analyte/internal standard was used to represent the contents of the variance analysis. The amounts of chemical markers were indicated by the ratio of compound and internal standard peak area. Results: By an overall analysis of RRLC-QTOF/MS, a total of 65 compounds were detected in the extracts from ACR in combination with TF, including 57 compounds with confirmed structures and 15 compounds with significant differences before and after combination, and the relative dissolution rates were affected by the changes of the combination ratios. Conclusion: The sensitive and accurate RRLC-QTOF/MS method could fully reflect the chemical constituent changes of ACR before and after combination with TF. On the basis of in vitro chemical constituent and contents, it is still unclear whether the combination of ACR and TF should be prohibited, which requires further in vivo investigation.