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.
Antioxidants/pharmacology* ; Chromatography, High Pressure Liquid/methods* ; Luteolin ; Plant Extracts/pharmacology* ; Trichosanthes/chemistry*

OBJECTIVETo investigate the effects of snakegourd root polysaccharide on apoptosis of human breast cancer cells (MCF-7 cells).

METHODSColorimetric MTT assay was used to measure the inhibition of snakegourd root polysaccharide on MCF-7 cells. The morphological changes of MCF-7 cells were observed by fluorescence microscope after DAPI staining and transmission electron microscope. The apoptosis of MCF-7 cells was examined by DNA agarose gel electrophoresis analysis of DNA fragmentation amd flow cytometry. The activity of Caspase-3 and Caspase-8 was detected by colorimetric assay.

RESULTSPolysaccharide of snakegourd root significantly inhibited MCF-7 cells in a dose-and time-dependent manner. The nuclear condensation and marginalization were observed by DAPI staining and transmission electron microscope. The characteristic ladder of apoptosis in DNA electrophoresis was detected in MCF-7 cells treated with 10.0 μmol/L polysaccharide of snakegourd root at d 2. The activities of Caspase-3 and Caspase-8 were increased in a time-dependent manner. The rates of apoptosis in MCF-7 cells were (5.2 ±1.3)%, (13.1 ±4.7)%, (27.6 ±6.8)% and (43.8 ±9.8)% treated with 1.0,5.0,10.0 and 20.0 μmol/L snakegourd root polysaccharide at d 2,respectively. The maximal activities of intracellular Caspase-3 and Caspase-8 were (2.32 ±0.12)U/μg and (1.92 ±0.11)U/μg at d 2 and d 1, respectively when MCF-7 cells were treated with 10.0 μmol/L.

CONCLUSIONThe polysaccharide of snakegourd root can induce the apoptosis of MCF-7 cells,which is associated with the activation of intracellular Caspase-3 and Caspase-8.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Humans ; MCF-7 Cells ; Plant Roots ; chemistry ; Polysaccharides ; pharmacology ; Trichosanthes ; chemistry

OBJECTIVETo compare and analyze volatile constituents from flowers of Trichosanthes kirilowii, in order to point out characteristic differences between female and male flowers.

METHODBlooming female and male flowers were collected in the same place. Volatile constituents were extracted from the flower by solid phase micro-extraction (SPME), then separated and analyzed by gas chromatography-mass-spectrometry (GC-MS).

RESULTFifty-two and forty-five chromatographic peaks were separated from the female and male flowers, respectively. Forty seven constituents were identified and their relative percentage compositions were determined with the peak area normalization method. Linalool, alpha-farnesene, benzene methanol, and (Z)-2-methylbutanal oxime were the main volatile constituents. The contents of linalool and alpha-farnesene in female flower were remarkably higher than those in male. In contrast, the content of benzene methanol in male flower was remarkably higher than that in female.

CONCLUSIONIn the first study on chemical constituents from flowers in genus Trichosanthes, 37 compounds are separated from T. kirilowii. Contents of linalool, alpha-farnesene and benzene methanol show the characteristic differences of volatile constituents contained in male and female flowers of T. kirilowii, which enriches the basic studies on dioecious plant.

Flowers ; chemistry ; Gas Chromatography-Mass Spectrometry ; Solid Phase Microextraction ; Trichosanthes ; chemistry ; Volatile Organic Compounds ; analysis ; chemistry ; isolation & purification

OBJECTIVETo explore the affinity relation among land race of Trichosanthes kirilowii and to provide evidence tor the classification and authentication.

METHODUsing scanning electron microscope, characteristics of the pericarp surface were studied comparatively.

RESULTThe pericarp surface of the cultivated Fructus Trichosanthis in Shandong showed the characteristics of generality and diversity.

CONCLUSIONThe results of the study could be used for the identification of the cultivated Fructus Trichosanthis in Shandong.

China ; Fruit ; ultrastructure ; Microscopy, Electron, Scanning ; Plants, Medicinal ; classification ; growth & development ; ultrastructure ; Quality Control ; Trichosanthes ; classification ; growth & development ; ultrastructure

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.
Animals ; Chromatography, High Pressure Liquid ; Delayed-Action Preparations ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Tandem Mass Spectrometry ; Trichosanthes

OBJECTIVETo investigate the cytotoxic effect of extracts of Trichosanthes kirilowi (TK) root on Hela cells in vitro and its probable anti-tumor mechanism.

METHODSThe cytotoxic effect in vitro on the growth of Hela cells was evaluated by microculture tetrazolium assay (MTT). Cell ultrastructural changes were examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and DNA agarose electrophoresis was performed to determine apoptosis and biochemical changes of Hela cells.

RESULTSExposure of Hela cells to TK extracts for 24-48 hrs resulted in a cell growth arrest, which showed in a time- and dose-dependent manner (r > 0.880, P < 0.01). With SEM and TEM, marked changes were observed, including microvilli disappearance or reduction, cell membrane vesiculation, cell shrinkage, condensation of chromosomes and apoptotic bodies with complete membrane. Besides, the apoptosis of Hela cells was confirmed by typical DNA ladder formation on gel electrophoresis.

CONCLUSIONExtracts of TK has a marked anti-tumor activity and could induce apoptosis of Hela cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Division ; drug effects ; DNA ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Plant Roots ; chemistry ; Trichosanthes ; chemistry

Smoke water and distillation liquid were used to treat the seeds of Trichosathes kirilowii and to study the effects of smoke water and distillation liquid on the seed germination and seedling growth of T. kirilowii. The results showed that germination rate, germination index and germination vigor of T. kirilowii all were significantly improved with the treatment of SW and DL treatment. The activity of α-amylase were significantly increased with the treatment of SW and DL at 1:2,000. SW and DL treatment showed no significant effects on the activity of SOD. The activity of POD were markedly enhanced under the treatment of SW (1:000) and DL (1:2,000). CAT activity were increased with the treatment of SW and DL at 1:2,000 while were inhibited by SW and DL at 1:500. Seedling height and root length were increased with the treatment of SW and DL (1:1,000, 1:2,000). SW and DL treaments improved the content of chlorophyll, and moreover with the concentration of SW and DL, the stimulatory were also increased. This work demonstrated that smoke water and diatillation liquid at 1:2,000 could stimulate the seed germination and seedling growth of T. kirilowii, and it provided the references for the study of seed germination technology.
Agriculture ; instrumentation ; methods ; Distillation ; Germination ; Seedlings ; growth & development ; metabolism ; Seeds ; growth & development ; metabolism ; Smoke ; analysis ; Trichosanthes ; growth & development ; metabolism ; Water ; chemistry ; metabolism

OBJECTIVETo study the chemical constituents of unsaponifiable matter from the seed oil of Trichosanthes hupehensis.

METHODThe fatty oil from the seeds of T. hupehensis was extracted with petroleum ether. The saponification was carried out with potassium hydroxide. The unsaponifiable matter was isolated and purified by silica gel column chromatography. Their structures were elucidated by means of MS, IR, UV, and 1H-NMR.

RESULTKarounidiol, isokarounidiol, 5-dehydrokarounidiol, 7-oxodihydrokarounidiol, stigmast-7-en-3beta-ol, stigmast-7, 22-dien-3beta-ol, 10alpha-Cucurbitadienol, beta-sitosterol, stigmast-7, 22-dien-3beta-O-beta-D-glucoside were elucidated.

CONCLUSIONAll of these compounds were found in this plant for the first time.

Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Seeds ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Trichosanthes ; chemistry ; Triterpenes ; chemistry ; isolation & purification

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.
Animals ; Area Under Curve ; Benzoates ; pharmacokinetics ; Chromatography, Liquid ; Delayed-Action Preparations ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Tandem Mass Spectrometry ; Trichosanthes ; chemistry

Trichosanthes kirilowii has been widely cultivated as its medicinal use, health care and food value. Drought resistance of seedlings is an important feature in breeding. Seeds of two T. kirilowii strains were used to research the difference of surface ultrastructure characteristic and drought resistance. Scanning electron microscope was used to identify the surface ultrastructure characteristic of seeds and PEG was used to simulate drought stress. The seeds germination rate, MDA content, chlorophyll content and the antioxidant enzymes activity were measured under the drought stress. The results showed that the seed surface colour of KXY-001 was lighter than that of KXY-005. The testa cobwebbing of KXY-001 was more intensive than that of KXY-005. The germination rate of KXY-001 was higher than that of KXY-005 under drought stress. The MDA content was increased and the chlorophyll content was decreased with the increasing of drought degree. The SOD activity of KXY-001 was higher than that of KXY-005, while the activity of POD and CAT was also increased firstly and decreased later. Surface reticulate of seeds and hilar traits can be used as identification points to identify the investigated strains. SOD and POD are activated to resist drought in T. kirilowii seedlings and the drought resistance of KXY-001 is superior than that of KXY-005.
Adaptation, Physiological ; drug effects ; Catalase ; metabolism ; Chlorophyll ; metabolism ; Droughts ; Germination ; Malondialdehyde ; metabolism ; Microscopy, Electron, Scanning ; Peroxidase ; metabolism ; Polyethylene Glycols ; pharmacology ; Seedlings ; metabolism ; Seeds ; growth & development ; metabolism ; ultrastructure ; Species Specificity ; Superoxide Dismutase ; metabolism ; Trichosanthes ; classification ; growth & development ; metabolism