Trigonelline protects the cardiocyte from hydrogen peroxide induced apoptosis in H9c2 cells

Objective:To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline(TG)for reducing the H_2O_2induced toxicity in H9c2 cells.Methods:Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit.RNA extraction and cDNA synthesized according to the kit manufacture protocol.Apoptosis was measured by the Flowcytometry,general PCR and qPCR.Results:It was found that the TG significantly rescued the morphology of the H9c2 cells.Treatment of cells with TG attenuated H_2O_2 induced cell deaths and improved the antioxidant activity.In addition,TG regulated the apoptotic gene caspase-3,caspase-9 and anti-apoptotic gene Bcl-2.Bcl-XL during H_2O_2 induced oxidative stress in H9c2 cells.These results were comparable with quercetin treatment.For evident,flow cytometer results also confirmed the TG significantly reduced the H_2O_2 induced necrosis and apoptosis in H9c2 cells.However,further increment of TG concentration against H_2O_2 could induce the necrosis and apoptosis along with H_2O_2.Conclusions:It is suggested that less than 125μM of TG could protect the cells from H_2O_2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression.Therefore,we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.

Protection effect of trigonelline on liver of rats with non-alcoholic fatty liver diseases

Objective: To study the effect of trigonelline on the change of indicators of serum transaminase, lipoprotein and liver lipid of model rats with non-alcoholic fatty liver diseases and on the expression level of Bcl-2 and Bax proteins. Methods: A total of 45 SD rats were randomly divided into Fthe control group, model group and trigonelline intervention group. Rats in the control group were fed with the common diet, while rats in the model group and intervention group were fed with the high fat diet. 8 weeks later, the intervention group received the intragastric administration of trigonellin e(with the dosage of 40 mg/kg/d) for 8 weeks; while control group and model group received the intragastric administration of saline with the equal dosage. Blood was taken from the abdominal aorta of rats 8 weeks later, detecting the level of a series of indicators of ALT, AST, TG, TC, HDL-C and LDL-C in the serum. After the rats were sacrificed, detect the indicators of TG, TC, SOD and MDA in the liver tissue of rats, as well as the expression of Bcl-2 and Bax in the liver tissue. Results: Results of histopathologic examination showed that the damage degree of liver for rats in the trigonellineintervention group was smaller than the one in the model group, with significantly reduced hepatic steatosis and the partially visible hepatic lobule. The levels of ALT, AST, TC and LDL-C in the serum of rats in the trigonelline group were significantly reduced, while the change in the levels of TG and HDL-C was not significantly different. The levels of TG, TC and MDA in the liver tissues were significantly decreased, while the level of SOD significantly increased; the expression of Bcl-2 protein in the liver tissues of rats in the trigonelline intervention group was significantly increased, while the expression of Bax protein significantly decreased. Conclusions: The trigonelline contributes to the therapeutic effect of non-alcoholic fatty liver diseases. It can also increase the expression of Bcl-2 protein and decrease the expression of Bax protein in the liver tissues, which can protect the liver.

Quantitative analysis of trigonelline in some Annona species by proton NMR spectroscopy

A quantitative ^(1)H NMR method(qHNMR)was used to measure the trigonelline content in the leaves of six species of the Annona genus.The methodology employed compared the intensities of the signals at δ9.14(H-2)andδ0.00,the internal standard TSP-d_(4).This measuring method was able to establish the concentration of trigonelline in the range from 0.67 to 10.04 mg·g^(-1) depending on the investigated extract.

Stress Responses of Peanut (<i>Arachis hypogaea</i>L.) Genotypes as Measured by Trigonelline Content after Exposure to UV-B Radiation

UV-B radiation has been widely documented as a stressor for plants that can cause decreased biomass, reduction in photosynthesis, and oxidative stress. Trigonelline is a secondary metabolite that is biosynthesized in some plants in response to abiotic stress such as UV-B irradiation. The objectives of this study were to examine biochemical stress responses for peanut plants (Arachis hypogaea L.) of four different genotypes (Spanish, Valencia, Virginia, and Runner) after exposure at various lengths to UV-B radiation and to examine the alteration of trigonelline biosynthesis due to the age of the plants. Peanut plants from the genotypes were exposed to UV-B radiation at three exposure times (60, 120, and 180 min);plants from two growth stages, the flowering (R1) and early maturity (R7), were used. Significant positive correlations (rs 0.29-0.74, P≤0.05) were found for trigonelline concentrations and UV-B exposure times. With longer exposure times of 180 min for plants at R7, trigonelline biosynthesis began as early as 10 days after treatment with 154.6 μg·g-1 DW and remained or increased by up to 71.5 μg·g-1 DW (46.3%) throughout the sampling intervals (10, 20, 30, 40, and 50 days after treatment) to a final value of 226.1 μg·g-1 DW. All four genotypes at R7 exhibited trigonelline concentrations 47.3% to 52.4% (71.6 to 96.5 μg·g-1 DW) higher than individuals at R1. Trigonelline biosynthesis at R7 was significantly (P<0.05) affected by all levels of UV-B exposure, whereas trigonelline concentrations at R1 were significantly influenced (P<0.05) by only the longer exposure times (120 and 180 min). No statistically significant difference was found in trigonelline concentration among the four different genotypes. UV-B irradiation had the greatest effect on plants at R7 after 120 and 180 min of exposure, as 15 out of 20 (75%) individuals had significantly higher (P<0.05) trigonelline concentrations.

Increase of Trigonelline in Trigonella persica Plant under Drought Stress

Trigonella persica is a valuable medicinal plant which comprises trigonelline that is secondary metabolite and important component in cosmetic and medicine.This research was conducted in order to evaluation the drought stress effect on growth parameters,root anatomical changes and trigonelline content in T.persica.Plants were grown under soil moisture corresponding to 100%,75%,50%and 25%field capacity for two weeks.The data showed that drought stress was significantly decreased fresh weight and dry weight of shoot and root.In addition,leaf area was declined due to drought stress.Interestingly,root length was enhanced by drought stress.Root microscopic study demonstrated that drought stress increased thickness of epidermal,endodermal,vascular bundle,central cylinder and parenchyma in T.persica.Drought stress caused a significant increment in alkaloid and trigonelline content in aerial parts and roots of T.persica.These results revealed that T.persica responded to drought stress by increasing the alkaloid and trigonelline,as well as the anatomical changes in root.Considering the importance of trigonline and alkaloids,this work may open prospects for production of the pharmaceutically valuable secondary metabolites thereby drought stress.

Antibacterial Effect of Aqueous Extracts and Bioactive Chemical Compounds of <i>Coffea canephora</i>against Microorganisms Involved in Dental Caries and Periodontal Disease

This study aimed to evaluate the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Coffea canephora aqueous extract (Cc), trigonelline (Tg) and 5-caffeoylquinic acid (5-CQA) against cariogenic microorganisms: Streptococcus parasanguinis ATCC 903 (SP), Lactobacillus rhamnosus ATCC 9595 (LR);and pathogens related to periodontal disease: Porphyromonas gingivalis ATCC 33277 (PG), Fusobacterium nucleatum ATCC 25586 (FN), Prevotella intermedia ATCC 49046 (PI) and Prevotella nigrescens ATCC 33563 (PN). Different concentrations of Cc (0.15625 to 10 mg/mL), Tg and 5-CQA (0.005 to 10.24 mg/mL) were tested. Chlorhexidine (0.05%) was used as positive control and the substances without the inoculum comprised the blank control. The Cc showed bacteriostatic action against SP (MIC = 5 mg/mL) and LR (MIC = 10 mg/mL). Tg showed bacteriostatic action against SP (MIC = 2.56 to 1.28 mg/mL), LR (MIC = 2.56 mg/mL), PG (MIC = 2.56 to 1.28 mg/mL), FN (MIC = 5.12 mg/mL), PN (MIC = 2.56 mg/mL), and PI (MIC = 2.56 to 1.28 mg/mL). Also, Tg showed bactericide properties against SP (MBC = 2.56 mg/mL), PG (MBC = 2.56 mg/mL), FN (MBC = 10.24 mg/mL), PN (MBC = 5.12 mg/mL), and PI (MBC = 2.56 mg/mL). Although 5-CQA has previously shown activity against Streptococcus mutans, in the present study, it showed no activity against all tested microorganisms. C. canephora extract only showed antibacterial activity against cariogenic microorganisms, not presenting action against periodontal pathogens. It was concluded that trigonelline presented the best effect against all pathogens tested, therefore coffee extracts with higher trigonelline content should be tested against these specific pathogens.