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1.
Bioorg Chem ; 149: 107487, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38805910

ABSTRACT

The peel of Trichosanthes kirilowii Maxim, is considered one of the primary sources for Trichosanthis pericarpium in traditional Chinese medicine, exhibiting lipid-lowering properties. The impact on hyperlipidemia mice of the crude polysaccharide from the peel of T. Kirilowii (TRP) was investigated in this study. The findings revealed that TRP exhibited a significant improvement in hepatic lipid deposition. Moreover, it significantly decreased serum levels of TC, TG, and LDL-C, while concurrently increasing HDL-C. 16S rRNA amplicon sequencing technique revealed that TRP group exhibited an increased relative abundance of Actinobacteria, a down-regulated relative abundance of Ruminiclostridium, and an up-regulated relative abundance of Ileibacterium. Therefore, TRP might play a role in anti-hyperlipidemia through regulation of the intestinal milieu and enhancement of microbial equilibrium. Consequently, targeted fractionation of TRP resulted in the isolation of a homogeneous acidic polysaccharide termed TRP-1. The TRP-1 polysaccharide, with an average molecular weight of 1.00 × 104 Da, and was primarily composed of Rha, GlcA, GalA, Glc, Gal and Ara. TRP-1 possessed a backbone consisting of alternating connections between â†’ 6)-α-Galp-(1 â†’ 4)-α-Rhap-(1 â†’ 6)-α-Galp-(2 â†’ 6)-ß-Galp-(1 â†’ 6)-α-Galp-(2 â†’ 6)-ß-Galp-(1 â†’ units and branched chain containing â†’ 6)-α-Glcp-(1→, 2,4)-ß-Glcp-(1, and â†’ 4)-α-GlapA-(1→. Both TRP and TRP-1 exhibited significant disruption of cholesterol micelles, highlighting their potential as lipid-lowering agents that effectively inhibit cholesterol absorption pathways.


Subject(s)
Cholesterol , Gastrointestinal Microbiome , Hyperlipidemias , Polysaccharides , Trichosanthes , Animals , Gastrointestinal Microbiome/drug effects , Trichosanthes/chemistry , Mice , Hyperlipidemias/drug therapy , Polysaccharides/pharmacology , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Cholesterol/metabolism , Cholesterol/blood , Hypolipidemic Agents/pharmacology , Hypolipidemic Agents/chemistry , Hypolipidemic Agents/isolation & purification , Male , Molecular Structure , Structure-Activity Relationship , Dose-Response Relationship, Drug
2.
J Ethnopharmacol ; 329: 118158, 2024 Jul 15.
Article in English | MEDLINE | ID: mdl-38614263

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Trichosanthis pericarpium (TP; Gualoupi, pericarps of Trichosanthes kirilowii Maxim) has been used in traditional Chinese medicine (TCM) to reduce heat, resolve phlegm, promote Qi, and clear chest congestion. It is also an essential herbal ingredient in the "Gualou Xiebai" formula first recorded by Zhang Zhongjing (from the Eastern Han Dynasty) in the famous TCM classic "Jin-Guì-Yào-Lüe" for treating chest impediments. According to its traditional description, Gualou Xiebai is indicated for symptoms of chest impediments, which correspond to coronary heart diseases (CHD). AIM OF THE STUDY: This study aimed to identify the antithrombotic compounds in Gualoupi for the treatment of CHD. MATERIALS AND METHODS: A CHD rat model was established with a combination of high-fat diet and isoproterenol hydrochloride (ISO) administration via subcutaneous multi-point injection in the back of the neck. This model was used to evaluate the antithrombotic effect of two mainstream cultivars of TP ("HaiShi GuaLou" and "WanLou") by analyzing the main components and their effects. Network pharmacology, molecular docking-based studies, and a zebrafish (Danio rerio) thrombosis model induced by phenylhydrazine was used to validate the antithrombosis components of TP. RESULTS: TP significantly reduced the body weight of the CHD rats, improved myocardial ischemia, and reduced collagen deposition and fibrosis around the infarcted tissue. It reduced thrombosis in a dose-dependent manner and significantly reduced inflammation and oxidative stress damage. Cynaroside, isoquercitrin, rutin, citrulline, and arginine were identified as candidate active TP compounds with antithrombotic effects. The key potential targets of TP in thrombosis treatment were initially identified by molecular docking-based analysis, which showed that the candidate active compounds have a strong binding affinity to the potential targets (protein kinase C alpha type [PKCα], protein kinase C beta type [PKCß], von Willebrand factor [vWF], and prostaglandin-endoperoxide synthase 1 [PTGS1], fibrinogen alpha [Fga], fibrinogen beta [Fgb], fibrinogen gamma [Fgg], coagulation factor II [F2], and coagulation factor VII [F7]). In addition, the candidate active compounds reduced thrombosis, improved oxidative stress damage, and down-regulated the expression of thrombosis-related genes (PKCα, PKCß, vWF, PTGS1, Fga, Fgb, Fgg, F2, and F7) in the zebrafish model. CONCLUSION: Cynaroside, isoquercitrin, rutin, citrulline, and arginine were identified as the active antithrombotic compounds of TP used to treat CHD. Mechanistically, the active compounds were found to be involved in oxidative stress injury, platelet activation pathway, and complement and coagulation cascade pathways.


Subject(s)
Coronary Disease , Fibrinolytic Agents , Molecular Docking Simulation , Network Pharmacology , Trichosanthes , Animals , Fibrinolytic Agents/pharmacology , Fibrinolytic Agents/therapeutic use , Fibrinolytic Agents/chemistry , Coronary Disease/drug therapy , Rats , Male , Trichosanthes/chemistry , Zebrafish , Rats, Sprague-Dawley , Disease Models, Animal , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Plant Extracts/therapeutic use , Medicine, Chinese Traditional/methods
3.
BMC Plant Biol ; 24(1): 286, 2024 Apr 16.
Article in English | MEDLINE | ID: mdl-38627660

ABSTRACT

Fruit length is a crucial agronomic trait of snake gourd (Trichosanthes anguina L); however, genes associated with fruit length have not been characterised. In this study, F2 snake gourd populations were generated by crossing the inbred lines, S1 and S2 (fruit lengths: 110 and 20 cm, respectively). Subsequently, bulk segregant analysis, sequencing, and fine-mapping were performed on the F2 population to identify target genes. Our findings suggest that the fruit length of snake gourd is regulated by a major-effect regulatory gene. Mining of genes regulating fruit length in snake gourd to provide a basis for subsequent selection and breeding of new varieties. Genotype-phenotype association analysis was performed on the segregating F2 population comprising 6,000 plants; the results indicate that the target gene is located on Chr4 (61,846,126-61,865,087 bp, 18.9-kb interval), which only carries the annotated candidate gene, Tan0010544 (designated TFL). TFL belongs to the MADS-box family, one of the largest transcription factor families. Sequence analysis revealed a non-synonymous mutation of base C to G at position 202 in the coding sequence of TFL, resulting in the substitution of amino acid Gln to Glu at position 68 in the protein sequence. Subsequently, an InDel marker was developed to aid the marker-assisted selection of TFL. The TFL in the expression parents within the same period was analysed using quantitative real-time PCR; the TFL expression was significantly higher in short fruits than long fruits. Therefore, TFL can be a candidate gene for determining the fruit length in snake gourd. Collectively, these findings improve our understanding of the genetic components associated with fruit length in snake gourds, which could aid the development of enhanced breeding strategies for plant species.


Subject(s)
Trichosanthes , Trichosanthes/genetics , Fruit/genetics , Plant Breeding , Phenotype , Genes, Plant/genetics
4.
Food Chem ; 443: 138547, 2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38271897

ABSTRACT

In the present study, we hypothesised that Trichosanthes kirilowii seed protein isolate (TPI) obtained by different extraction methods have distinct structure, functional attributes and volatile profiles. Alkaline-extracted isolate (AE-TPI) exhibited lower protein content and a darker colour than the other two isolates because more polyphenols and pigments were coextracted. Salt-extracted isolate (SE-TPI) and AE-TPI had higher in vitro protein digestibility than reverse micelle-extracted isolate (RME-TPI) due to higher degrees of denaturation, which enabled them to be more susceptible to proteolysis. The SE-TPI gel resulted in a stronger gel network and greater hardness than the other two isolate gels. In the volatile profile, SE-TPI (22) yielded the largest number of volatile compounds, followed by AE-TPI (20) and RME-TPI (15). The current results indicated that the structure, functional properties and volatile profiles of TPI are largely influenced by the extraction technique.


Subject(s)
Trichosanthes , Trichosanthes/chemistry , Seeds/chemistry
5.
Nutr Cancer ; 75(10): 1945-1957, 2023.
Article in English | MEDLINE | ID: mdl-37870977

ABSTRACT

The roots of Trichosanthes kirilowii (TK) have been used in traditional oriental medicine for the treatment of respiratory diseases. In this study, we investigated whether an ethanolic root extract of TK (ETK) can regulate the metastatic potency of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI)-resistant human lung cancer cells. The relative migration and invasion abilities of erlotinib-resistant PC9 (PC9/ER) and gefitinib-resistant PC9 (PC9/GR) cells were higher than those of parental PC9 cells. Mesenchymal markers were overexpressed, whereas epithelial markers were downregulated in resistant cells, suggesting that resistant cells acquired the EMT phenotype. ETK reduced migration and invasion of resistant cells. The expression levels of N-cadherin and Twist were downregulated, whereas Claudin-1 was upregulated by ETK, demonstrating that ETK suppresses EMT. As a molecular mechanism, Src was dephosphorylated by ETK. The anti-metastatic effect of ETK was reduced by transfecting PC9/ER cells with a constitutively active form of c-Src. Dasatinib downregulated N-cadherin, Twist, and vimentin, suggesting that Src regulates EMT in resistant cells. Notably, CuB played a key role in mediating the anti-metastatic activity of ETK. Collectively, our results demonstrate that ETK can attenuate the metastatic ability of EGFR-TKI-resistant lung cancer cells by inhibiting Src-mediated EMT.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Trichosanthes , Humans , Lung Neoplasms/metabolism , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/metabolism , Trichosanthes/metabolism , ErbB Receptors/genetics , ErbB Receptors/metabolism , Protein Kinase Inhibitors/pharmacology , Drug Resistance, Neoplasm , Cell Line, Tumor , Cadherins
6.
Virology ; 588: 109891, 2023 11.
Article in English | MEDLINE | ID: mdl-37826911

ABSTRACT

Trichosanthes kirilowii has been mainly grown for use in traditional Chinese medicine. In this study, cucurbit mild mosaic virus (CuMMV) belonging to the genus Fabavirus was identified from T. kirilowii plants. CuMMV possesses a segmented, bipartite linear single-stranded RNA genome composed of RNA1 and RNA2. Sequence analysis showed that each genomic segment shares the highest sequence similarity with those of CuMMV isolated from pumpkin. A full-length infectious cDNA clone of CuMMV was further constructed and was found to induce typical symptoms in T. kirilowii, Cucumis sativus, C. melo, Citrullus lanatus, and Cucurbita pepo. The sap inoculum derived from the infectious cDNA clone of CuMMV could be mechanically transmitted and reproduce similar symptoms in the tested plants. This is the first report on the construction of a biologically active, full-length infectious cDNA clone of CuMMV, which will provide a useful tool in understanding CuMMV-encoded proteins and plant-CuMMV interactions.


Subject(s)
Cucumis sativus , Fabavirus , Mosaic Viruses , Trichosanthes , Trichosanthes/genetics , DNA, Complementary/genetics , Fabavirus/genetics , Cucumis sativus/genetics , Plants , Mosaic Viruses/genetics
7.
Bioorg Chem ; 139: 106710, 2023 10.
Article in English | MEDLINE | ID: mdl-37418785

ABSTRACT

The pericarps of Trichosanthes kirilowii are often used to treat cough in traditional Chinese medicine, and its ethanol extract exhibited effective therapeutic effects on acute lung injury (ALI) in vivo caused by H1N1. An anticomplement activity-guided fractionation on the extract resulted in the isolation of ten new terpenoids, including seven monoterpenoids, trichosanates A-G (1-7), and three cucurbitane-type triterpenoids, cucurbitacins W-Y (8-10), as well as eleven known terpenoids (11-21). The new terpenoids' structures were determined by spectroscopic analysis, X-ray crystallographic analysis (1), electronic circular dichroism (ECD) analysis and calculations (2-10). Twelve monoterpenoids (1-7 and 11-15) and five cucurbitane-type triterpenoids (8-10, 18, and 20) exhibited anticomplement activity in vitro. For the monoterpenoids, the long aliphatic chain substituents might enhance their anticomplement activity. Additionally, two representative anticomplement terpenoids, 8 and 11, obviously attenuated H1N1-induced ALI in vivo by inhibiting complement overactivation and reducing inflammatory responses.


Subject(s)
Influenza A Virus, H1N1 Subtype , Trichosanthes , Triterpenes , Cucurbitacins , Trichosanthes/chemistry , Monoterpenes/pharmacology , Triterpenes/pharmacology , Triterpenes/chemistry , Plant Extracts/pharmacology
8.
BMC Plant Biol ; 23(1): 354, 2023 Jul 10.
Article in English | MEDLINE | ID: mdl-37430217

ABSTRACT

BACKGROUND: The APETALA 2/ ethylene-responsive element binding factors (AP2/ERF), are thought to be associated with plant abiotic stress response, and involved in some plant hormone signaling pathways. Trichosanthes kirilowii is an important edible and medicinal crop, so far no research has been conducted on the TkAP2/ERF genes. RESULT: In this study, a total of 135 TkERFs were identified, these genes were divided into 4 subfamilies and clustered into 13 groups. Moreover, 37 paralogous pairs were identified, with only two having Ka/Ks values greater than 1, proving that most TkERF genes underwent purifying selection during evolution. Co-expression networks constructed using transcriptome data at various flowering stages revealed that 50, 64, and 67 AP2/ERF genes correlated with members of the ethylene, gibberellin, and abscisic acid signaling pathways, respectively. When tissue cultured seedlings were treated with ETH, GA3 and ABA, 11, 12 and 17 genes were found to be up-regulated, respectively, suggesting that some members of the TkERF gene family may be involved in plant hormone signaling pathways. And under 4 ℃, PEG and NaCl treatment, 15, 20 and 19 genes were up-regulated, respectively, this suggested that these selected genes might be involved in plant abiotic stresses. CONCLUSIONS: Overall, we identified 135 AP2/ERF family members, a comprehensive analysis of AP2/ERF gene expression patterns by RNA-seq and qRT-PCR showed that they played important roles in flower development and abiotic stress. This study provided a theoretical basis for the functional study of TkAP2/ERF genes and the genetic improvement of T. kirilowii.


Subject(s)
Trichosanthes , Plant Growth Regulators/pharmacology , Ethylenes , Gibberellins , Polymerase Chain Reaction
9.
Curr Pharm Biotechnol ; 24(13): 1694-1707, 2023.
Article in English | MEDLINE | ID: mdl-36856179

ABSTRACT

BACKGROUND: The treatment of diabetes with plant ingredients such as in Traditional Chinese Medicine (TCM) is an alternative to classical chemotherapy. OBJECTIVE: This study aims to explore the hypoglycemic effect of Trichosanthes pericarpium powder (TP) and intestinal bacteria transplantation in type 2 diabetic mice. The relationship between intestinal bacteria transplantation and improvement in insulin resistance was also investigated. METHODS: The polyphenols and terpenoids in the TP were identified by LC-MS/MS. Streptozotocin was used to induce a mouse model of type 2 diabetes. Diabetic mice were treated with different doses of TP and the intestinal bacteria obtained from the high-dose TP group for four weeks. RESULTS: As a result, FBG levels were found to be significantly reduced in diabetic mice, weight gain and organ enlargement were alleviated, and insulin resistance was significantly improved. TP administration also improved the disorder in intestinal bacteria in diabetic mice. Besides, TP can increase the liver AMPK, SIRT1, GLUT1, and GLUT4 gene expression, while down-regulated PEPCK and G6Pase gene expression suggest a potential mechanism for hypoglycemia in diabetic mice. CONCLUSION: Oral administration of Trichosanthes pericarpium powder in the treatment of diabetes may be achieved by restoring hepatic function, improving insulin resistance, and the dynamic balance of intestinal bacteria.


Subject(s)
Diabetes Mellitus, Experimental , Diabetes Mellitus, Type 2 , Insulin Resistance , Trichosanthes , Mice , Animals , Hypoglycemic Agents/pharmacology , Hypoglycemic Agents/therapeutic use , Diabetes Mellitus, Type 2/drug therapy , Trichosanthes/metabolism , Diabetes Mellitus, Experimental/drug therapy , Chromatography, Liquid , Powders/metabolism , Powders/pharmacology , Powders/therapeutic use , Blood Glucose/metabolism , Tandem Mass Spectrometry , Bacteria/metabolism , Insulin/metabolism , Liver
10.
J Vis Exp ; (193)2023 03 03.
Article in English | MEDLINE | ID: mdl-36939253

ABSTRACT

We aimed to study the mechanism of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma (LUAD) based on network pharmacology and experimental verification. The effective components and potential targets of Trichosanthis and Fritillaria thunbergii were collected by high-throughput experiment and reference-guided (HERB) database of traditional Chinese medicine and a similarity ensemble approach (SEA) database, and the LUAD-related targets were queried by the GeneCards and Online Mendelian Inheritance in Man (OMIM) databases. A drug-component-disease-target network was constructed by Cytoscape software. Protein-protein interaction (PPI) network, gene ontology (GO) function, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted to obtain core targets and key pathways. An aqueous extract of Trichosanthes-Fritillaria thunbergii and A549 cells were used for the subsequent experimental validation. Through the HERB database and literature search, 31 effective compounds and 157 potential target genes of Trichosanthes-Fritillaria thunbergii were screened, of which 144 were regulatory targets of Trichosanthes-Fritillaria thunbergii in the treatment of lung adenocarcinoma. The GO functional enrichment analysis showed that the mechanism of action of Trichosanthes-Fritillaria thunbergii against lung adenocarcinoma is mainly protein phosphorylation. The KEGG pathway enrichment analysis suggested that the treatment of lung adenocarcinoma by Trichosanthes-Fritillaria thunbergii mainly involves the PI3K/AKT signaling pathway. The experimental validation showed that an aqueous extract of Trichosanthes-Fritillaria thunbergii could inhibit the proliferation of A549 cells and the phosphorylation of AKT. Through network pharmacology and experimental validation, it was verified that the PI3K/AKT signaling pathway plays a vital role in the action of Trichosanthes-Fritillaria thunbergii in treating lung adenocarcinoma.


Subject(s)
Adenocarcinoma of Lung , Fritillaria , Lung Neoplasms , Trichosanthes , Humans , Network Pharmacology , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , Adenocarcinoma of Lung/drug therapy , Databases, Genetic , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Molecular Docking Simulation
11.
BMC Genomics ; 24(1): 81, 2023 Feb 21.
Article in English | MEDLINE | ID: mdl-36810020

ABSTRACT

Trichosanthes kirilowii Maxim. (TK) is a dioecious plant in the Cucurbitaceae family of which different sexes have separate medicinal uses. We used Illumina high-throughput sequencing technology to sequence miRNAs from male and female flower buds of TK. We performed bioinformatics analysis, miRNA identification, and target gene prediction on the data obtained from sequencing, and association analysis was performed in combination with the results of a previous transcriptome sequencing study. As a result, there were 80 differentially expressed miRNAs (DESs) between the female and male plants (48 upregulated and 32 downregulated in female plants). Moreover, 27 novel miRNAs in DESs were predicted to have 282 target genes, and 51 known miRNAs were predicted to have 3418 target genes. By establishing a regulatory network between miRNAs and target genes, 12 core genes were screened, including 7 miRNAs and 5 target genes. Among them, tkmiR157a-5p, tkmiR156c, tkmiR156_2, and tkmiR156k_2 jointly target the regulation of tkSPL18 and tkSPL13B. These two target genes are specifically expressed in male and female plants, respectively, and are involved in the biosynthesis process of BR, which is closely related to the sex differentiation process of TK. The identification of these miRNAs will provide a reference for the analysis of the sex differentiation mechanism of TK.


Subject(s)
Cucurbitaceae , MicroRNAs , Trichosanthes , Trichosanthes/genetics , MicroRNAs/genetics , Gene Expression Regulation, Plant , Cucurbitaceae/genetics , High-Throughput Nucleotide Sequencing/methods
12.
Nutr Cancer ; 75(1): 376-387, 2023.
Article in English | MEDLINE | ID: mdl-36004720

ABSTRACT

The aim of this study was to investigate whether the ethanol extract of the Trichosanthes kirilowii root (ETK), traditionally used to treat lung diseases, exhibits anticancer activity in epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI)-resistant non-small cell lung cancer (NSCLC) cells. ETK treatment suppressed the growth of EGFR TKI-resistant NSCLC cells, including H1299, H1975, PC9/ER (erlotinib-resistant PC9) and PC9/GR (gefitinib-resistant PC9) cells, in a concentration- and time-dependent manner. Dose-dependent decline in anchorage-dependent and -independent colony formation was also detected following ETK treatment. We demonstrate that the growth-inhibitory effect of ETK was related to apoptosis induction, based on flow cytometry results showing ETK-induced increase in the percentage of cells with sub-G1 DNA and the population of annexin V-positive cells. Consistently, ETK induced chromatin condensation and cleavage of poly(ADP-ribose) polymerase (PARP). As a molecular mechanism, the phosphorylation level of signal transducer and activator of transcription 3 (STAT3) and Src was decreased by ETK. ETK-induced apoptosis was partially reversed by transfection of constitutively activated STAT3, indicating that STAT3 inactivation mediated ETK-induced apoptosis in EGFR TKI-resistant NSCLC cells. Our results provide basic evidence supporting the role of ETK as a novel therapeutic in EGFR TKI-resistant NSCLC.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Plant Extracts , Humans , Apoptosis , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/metabolism , Cell Line, Tumor , Cell Proliferation , Drug Resistance, Neoplasm , ErbB Receptors/genetics , ErbB Receptors/metabolism , Lung Neoplasms/drug therapy , Lung Neoplasms/metabolism , Plant Extracts/pharmacology , Trichosanthes/chemistry
13.
Molecules ; 27(20)2022 Oct 18.
Article in English | MEDLINE | ID: mdl-36296619

ABSTRACT

Trichosanthes anguina L. (family Cucurbitaceae) is a monoecious and diclinous plant that can be consumed as a vegetable and has anti-inflammatory and antioxidant effects. The chemical composition and content of volatile compounds in female and male buds of T. anguina were explored by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) technology combined with multivariate statistical analysis. The results showed that the content of the volatile compounds was different between female and male buds. 2,2,6-trimethyl-6-vinyltetrahydro-2H-pyran-3-ol and 2,2,6-trimethyl-6-vinyldihydro-2H-pyran-3(4H)-one were the main volatile compounds in both female and male buds. Based on the multivariate statistical analysis of orthogonal projections to latent structures discriminant analysis (OPLS-DA) and t-test, the content of seven compounds was significantly different between female and male buds. The content of three compounds in male buds was higher than that in female, i.e., (E)-4,8-dimethyl-1,3,7-nonatriene, 1,5,9,9-tetramethyl-1,4,7-cycloundecatriene, and (E)-caryophyllene. Conversely, the content of (Z)-4-hexen-1-ol, (Z)-3-hexenyl benzoate, (Z)-3-hexenyl salicylate, and 2-hexen-1-ol in female buds was higher than that in male buds. This is the first report on the difference in the volatile compounds between female and male buds of T. anguina, which enriches the basic research on the monoecious and diclinous plant and provides a reference for the study of plant sex differentiation.


Subject(s)
Trichosanthes , Volatile Organic Compounds , Solid Phase Microextraction/methods , Gas Chromatography-Mass Spectrometry/methods , Antioxidants/analysis , Volatile Organic Compounds/analysis , Pyrans/analysis
14.
Planta ; 256(3): 58, 2022 Aug 18.
Article in English | MEDLINE | ID: mdl-35980476

ABSTRACT

MAIN CONCLUSION: Two key amino acids of isomultiflorenol synthase, Y125 and M254, were first proposed. They could be associated with the production of isomultiflorenol. Oxidosqualene cyclases (OSCs) are the first committed enzymes in the triterpenoid biosynthesis by converting 2,3-oxidosqualene to specific triterpenoid backbones. Thus, these enzymes are potential targets for developing plant-active compounds through the study of triterpenoid biosynthesis. We applied transcriptome information and metabolite profiling from Trichosanthes cucumerina L. to define the diversity of triterpenoids in this plant through OSCs. Isomultiflorenol synthase and cucurbitadienol synthase were previously identified in this plant. Here, three new OSCs, TcBAS, TcLAS, and TcCAS, were cloned and functionally characterized as ß-amyrin synthase, lanosterol synthase, and cycloartenol synthase activities, respectively. We also took advantage of the multiple sequence alignment and molecular docking of OSCs exhibiting in this plant and other plant OSCs to identify key residues associated with isomultiflorenol synthase specificity. Two novel key amino acids, referred to the Y125 and M254, were first discovered. These results provide information on a possible catalytic mechanism for plant OSCs that produce specific products.


Subject(s)
Intramolecular Transferases , Trichosanthes , Triterpenes , Amino Acids , Cloning, Molecular , Intramolecular Transferases/genetics , Intramolecular Transferases/metabolism , Molecular Docking Simulation , Squalene/analogs & derivatives , Substrate Specificity , Trichosanthes/metabolism , Triterpenes/metabolism
15.
Genet Res (Camb) ; 2022: 1792977, 2022.
Article in English | MEDLINE | ID: mdl-35919037

ABSTRACT

Background: Oxidative stress is an important cause of liver disease and atherosclerosis. Natural substances with antioxidant activity are good drugs for treating liver disease and atherosclerosis. Trichosanthes kirilowii Peel Polysaccharide (TKPP) can remove DPPH (2,2-Diphenyl-1-picrylhydrazyl) free radicals and hydroxyl free radicals in vitro, which shows antioxidant activity. Therefore, it is speculated that it can protect human hepatoma cell line (HepG2) and umbilical artery smooth muscle cell (HUASMC) against oxidative damage by hydrogen peroxide (H2O2). Methods: Oxidative damage cell models of HepG2 and HUASMC were induced by H2O2. HepG2 and HUASMC were divided into blank group, H2O2 injury group, TKPP treatment group, and glutathione (GSH) positive control group. Cell Counting Kit-8 (CCK-8) was used to detect cell viability. The level of total GSH and the amount of Nitric oxide (NO) secreted by cells were detected by specific kits. The gene and protein expressions of catalase (CAT) and superoxide dismutase (SOD) were detected by fluorescence quantitative PCR and Western Blot. Results: In these two kinds of cells, compared with the control group, the survival rate, total GSH level, and NO secretion, CAT and SOD gene and protein expressions were significantly decreased in the H2O2 damaged group. In the TKPP treatment group, the cell survival rate was significantly elevated with the increase of the polysaccharide concentration, and the total GSH level, NO secretion, CAT and SOD gene expression, and protein expression levels were also significantly increased. Conclusion: TKPP can improve the activities of HepG2 and HUASMC cells damaged by H2O2 and protect the cellular antioxidant system.


Subject(s)
Atherosclerosis , Trichosanthes , Antioxidants/metabolism , Antioxidants/pharmacology , Glutathione/metabolism , Glutathione/pharmacology , Humans , Hydrogen Peroxide/toxicity , Oxidative Stress , Polysaccharides/pharmacology , Superoxide Dismutase/metabolism , Superoxide Dismutase/pharmacology , Trichosanthes/metabolism
16.
Biotechnol Lett ; 44(10): 1127-1138, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35925526

ABSTRACT

Trichosanthes kirilowii Maxim taxonomically belongs to the Cucurbitaceae family and Trichosanthes genus. Its whole fruit, fruit peel, seed and root are widely used in traditional Chinese medicines. A ribosome-inactivating protein with RNA N-glycosidase activity called Trichosanthrip was isolated and purified from the seeds of T. kirilowii in our recent previous research. To further explore the biological functions of Trichosanthrip, the cDNA of T. kirilowii alpha-amylase inhibitor (TkAAI) was cloned through rapid-amplification of cDNA ends and its sequence was analyzed. Also, the heterologous protein was expressed in Escherichia coli and its alpha-amylase activity was further measured under optimized conditions. The full-length cDNA of TkAAI was 613 bp. The speculated open reading frame sequence encoded 141 amino acids with a molecular weight of 16.14 kDa. Phylogenetic analysis demonstrated that the Alpha-Amylase Inhibitors Seed Storage domain sequence of TkAAI revealed significant evolutionary homology with the 2S albumin derived from the other plants in the Cucurbitaceae group. In addition, TkAAI was assembled into pET28a with eGFP to generate a prokaryotic expression vector and was induced to express in E. coli. The TkAAI-eGFP infusion protein was proven to exhibit alpha-amylase inhibitory activity against porcine pancreatic amylase in a suitable reaction system. Analysis of gene expression patterns proved that the relative expression level of TkAAI in seeds is highest. The results presented here forecasted that the TkAAI might play a crucial role during the development of T. kirilowii seeds and provided fundamental insights into the possibility of T. kirilowii derived medicine to treat diabetes related diseases.


Subject(s)
Trichosanthes , Albumins , Amino Acids , Amylases , Animals , Cloning, Molecular , DNA, Complementary/genetics , Escherichia coli/genetics , Phylogeny , Saporins , Swine , Trichosanthes/chemistry , Trichosanthes/genetics , alpha-Amylases/genetics
17.
Food Res Int ; 159: 111643, 2022 09.
Article in English | MEDLINE | ID: mdl-35940816

ABSTRACT

Trichosanthes kirilowii Maxim seed is a primary source of edible vegetable oil and possesses a high nutritional value, making them extremely beneficial to humanity. To promote the extraction process of Trichosanthes kirilowii Maxim seed oil, the effect of microwave heating time (700 W for 0, 2, 4, and 6 min) on lipid composition, chemical properties, and antioxidant activity of oils was studied. The results showed that the oil yield of the seed increased with the microwave heating time. Besides, microwave heating time significantly affects (p < 0.05) DPPH and tocopherols, and the IC50 value of DPPH was highest with microwave heating for 6 min, whatever the shells are reserved. The tocopherol content was highest with microwave heating for 2 min in the seed shell oil, which was 1930.60 mg/kg. The longer microwave heating time could improve the oil yield and antioxidant activity of Trichosanthes kirilowii Maxim seed oil. The seed shell also affects chemical properties, fatty acid composition, antioxidant activity, and tocopherol contents of the Trichosanthes kirilowii Maxim seed oil. The Trichosanthes kirilowii Maxim seed shell oil has higher DPPH and tocopherols contents than seed kernel oil, while seed kernel oils showed higher oil yield and acid value. Our finding is valuable for manufacturers to choose suitable means to produce Trichosanthes kirilowii Maxim seed oil of required qualities and chemical compositions for targeted use.


Subject(s)
Trichosanthes , Antioxidants/analysis , Heating , Microwaves , Plant Oils/analysis , Seeds/chemistry , Tocopherols/analysis , Trichosanthes/chemistry
18.
J Oleo Sci ; 71(6): 823-827, 2022.
Article in English | MEDLINE | ID: mdl-35661064

ABSTRACT

Trichosanthes cucumerina seed and seed oil have been investigated for their chemical, physico-chemical properties, characterization and fatty acid profile. The properties were assessed by standard methods and the fatty acid profile was carried out using gas chromatography fatty acid methyl ester analysis. The seed was found to contain 71.1% crude oil, 1.6% Crude Protein, 2.9% carbohydrate, 2.8% crude Fiber, and 1.6% Ash. Iodine, Peroxide, and Acid the molecular weights of the seed oil are 0.84 gI2 of oil, 2.0 meq O2/kg oil and 4.0 mg/KOH/g, respectively, with 4.0 Free Fatty Acid and a Saponification Value of 379.12 mg/g. The oil is rich in polyunsaturated fatty acids that are vital for growth, maintenance of the cell membrane, development, and boosting of the immune system in humans. It qualifies to be classified as an edible vegetable oil and can also be useful in the soap and cosmetic industry.


Subject(s)
Trichosanthes , Fatty Acids/analysis , Fatty Acids, Nonesterified/analysis , Humans , Plant Oils/chemistry , Seeds/chemistry
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 42(3): 384-391, 2022 Mar 20.
Article in Chinese | MEDLINE | ID: mdl-35426802

ABSTRACT

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.


Subject(s)
Trichosanthes , Antioxidants/pharmacology , Chromatography, High Pressure Liquid/methods , Luteolin , Plant Extracts/pharmacology , Trichosanthes/chemistry
20.
J Diabetes Res ; 2022: 3250016, 2022.
Article in English | MEDLINE | ID: mdl-35224106

ABSTRACT

This study investigates the effects of the water-soluble and organic-soluble Trichosanthes extracts on the hyperglycemic condition in streptozotocin- (STZ-) induced diabetic rats. The blood glucose levels, body weights, water intake, and urine volumes of rats in different experimental groups were monitored throughout the experiment, and the results obtained indicate that the two extracts can effectively reduce blood sugar levels, increase body weights, and improve water intake and urine volumes in diabetic rats. Based on blood biochemical analyses, the two extracts play an important role in regulating the diabetes-induced lipid metabolism disorder, increasing the levels of insulin and C-peptide, and alleviating the symptoms of diabetes. The variation in the liver glycogen contents of the water-soluble fraction and ethanol fraction groups suggests that the mechanisms underlying the hypoglycemic effects of the two extracts are different. Indeed, the water-soluble fraction alleviates diabetes symptoms in rats mainly by antioxidative activity, unlike the ethanol fraction.


Subject(s)
Diabetes Mellitus, Type 1/drug therapy , Plant Extracts/metabolism , Trichosanthes/metabolism , Animals , Antioxidants/metabolism , Antioxidants/pharmacology , Antioxidants/therapeutic use , Blood Glucose/metabolism , Diabetes Mellitus, Type 1/physiopathology , Disease Models, Animal , Hypolipidemic Agents/metabolism , Hypolipidemic Agents/pharmacology , Hypolipidemic Agents/therapeutic use , Insulin/blood , Insulin/metabolism , Insulin/therapeutic use , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Rats
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