Animal studies were approved by the Institution’s Animal Care and Use Committee, and they conform to the US Department of Agriculture regulations and the NIH’s Guide for human care and use of Laboratory Animals. Male C57BL/6 wild-type mice at 8 weeks old received intraperitoneal injection of low-dose streptozotocin (55 mg/kg, STZ, Sigma-Aldrich). STZ injection was repeated daily for 5 consecutive days. Mice treated with sodium citrate buffer served as non-diabetic controls. One week after the final STZ injection, mice with nonfasting blood glucose of less than 280 mg/dl were culled, as they would unlikely develop sufficient diabetes to cause significant renal injury. Beginning 4 weeks after STZ injections, mice were randomized to receive daily intraperitoneal injection with β-HB (100 mg/kg/day) or vehicle in the presence or absence of intraperitoneal injection of trigonelline (1 mg/kg, Sigma-Aldrich) every other day for 4 weeks. The dose of β-HB was chosen according to previous studies18 (link) and optimized by pilot experiments. Six mice were randomly assigned to each group. Blood and spot urine were collected at indicated time points. Mice were euthanized at 8 weeks after the STZ injection.
Trigonelline
Manufactured by Merck Group
Sourced in United States, Germany
Trigonelline is a chemical compound that occurs naturally in various plants, including coffee beans. It is a water-soluble vitamin B3 derivative and has a crystalline structure. Trigonelline is commonly used as a reference standard in analytical laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Caffeine, by Merck Group (8 mentions)
Chlorogenic acid, by Merck Group (6 mentions)
Formic acid, by Merck Group (5 mentions)
Acetonitrile, by Merck Group (4 mentions)
Caffeic acid, by Merck Group (2 mentions)
Dithiothreitol (dtt), by Merck Group (2 mentions)
D glucose, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Quercetin, by Merck Group (2 mentions)
Protoporphyrin znpp, by Santa Cruz Biotechnology (2 mentions)
Cinnamic acid, by Merck Group (2 mentions)
Gallic acid, by Merck Group (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Cisplatin, by Merck Group (2 mentions)
Theobromine, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
N methyl pyrrolidone (nmp), by Merck Group (1 mentions)
Niacin, by Merck Group (1 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
27 protocols using trigonelline
1
Streptozotocin-Induced Diabetic Mouse Model
2
Coffee Capsule Composition Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Capsules from sixty-five different types of coffee capsules (named 1 to 65) belonging to five different brands (named A to E), together with their relative coffee machine, were purchased on local markets in Parma or through online stores during 2016 (23 capsules, named A1-A23, from brand A; 15 capsules, B24-B38, brand B; 10 capsules, C39-C48, brand C; 10 capsules, D49-D58, brand D; and 7 capsules, E59-E65, brand E). Two lots for each type of capsule were purchased. The type of coffee – regular, large (lungo) or decaffeinated EC – and the amount of coffee powder for each capsule is provided in Supplementary Table 1 .
3-O-Caffeoylquinic acid (3-CQA), 4-O-caffeoylquinic acid (4-CQA), 5-O-caffeoylquinic acid (5-CQA), caffeine, trigonelline, NMP, and niacin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Acetonitrile and formic acid were also purchased from Sigma-Aldrich. Water for UHPLC analysis was purchased from VWR Chemicals (Fontenay-sous-bois, France).
3-O-Caffeoylquinic acid (3-CQA), 4-O-caffeoylquinic acid (4-CQA), 5-O-caffeoylquinic acid (5-CQA), caffeine, trigonelline, NMP, and niacin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Acetonitrile and formic acid were also purchased from Sigma-Aldrich. Water for UHPLC analysis was purchased from VWR Chemicals (Fontenay-sous-bois, France).
3
Tamoxifen Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tamoxifen was purchased from Nacalai Tesque (Tokyo, Japan). Caffeine, caffeic acid, chlorogenic acid, pyrocatechol, and trigonelline were purchased from Sigma-Aldrich (St. Louis, MO). LY294002 and U0126 were purchased from Tocris Bioscience (Bristol, UK). Anti-ERα, anti-phospho-MEK1/2 (S217/221), anti-MEK1/2, anti-phospho-ERK1/2 (T202/Y204), anti-ERK1/2, anti-phospho-Akt (S473), anti-Akt, and anti-cleaved caspase-3 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-p53, anti-cyclin D1, and anti-β-actin antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Peroxidase-conjugated rabbit anti-mouse, goat anti-rabbit, and swine anti-goat secondary antibodies were from Dako (Glostrup, Denmark).
4
Preparation of Standard Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Trigonelline (analytical standard) was purchased
from Sigma-Aldrich (St Louis, MO); caffeine (99% purity), genistein
(99% purity), and theobromine (99% purity) were purchased from Alfa
Aesar by Thermo Fisher Scientific (Kandel, Germany). Stock solutions
of all compounds were prepared in mQ H2O at a concentration
of 1 mg/mL and stored at −20 °C. All other reagents were
purchased from Sigma-Aldrich (St Louis, MO) unless otherwise indicated.
from Sigma-Aldrich (St Louis, MO); caffeine (99% purity), genistein
(99% purity), and theobromine (99% purity) were purchased from Alfa
Aesar by Thermo Fisher Scientific (Kandel, Germany). Stock solutions
of all compounds were prepared in mQ H2O at a concentration
of 1 mg/mL and stored at −20 °C. All other reagents were
purchased from Sigma-Aldrich (St Louis, MO) unless otherwise indicated.
5
Pisum sativum Antioxidant Enzyme Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The voucher specimens (Pisum sativum L. var. saccharatum Poir: CMU-104-PS-003) were deposited in Herbarium of College of Pharmacy, China Medical University, Taichung, Taiwan. Antipain, aprotinin, dithiothreitol, ethyleneglycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid, leupeptin, Nonidet P-40, pepstatin, phenylmethylsulfonyl fluoride, sodium deoxycholate, trigonelline, and 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris) were purchased from Sigma Chemical Company (St. Louis, MO). Antibodies to various proteins were obtained from the following sources: β-actin antibody was purchased from Sigma Chemical Company; Caspase-9, catalase, p38 (pThr180/Tyr182), and Raf (pSer259) were purchased from Abcam (Cambridge, MA); Mn-SOD and Cu/Zn-SOD were from Calbiochem (San Diego, CA); ERK (pThr202/Tyr204) was from ThermoFisher Scientific, Inc. (Waltham, MA); Nrf2 (pSer40) was from GeneTex, Inc. (Irvine, CA); Caspase-3 and protein kinase Cα (PKCα) from BD Biosciences (San Jose, CA). Horseradish peroxidase (HRP)-conjugated goat anti-mouse and -rabbit IgG were from Abcam.
6
Corticosterone-Induced Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), poly-L-lysine, corticosterone (CORT), trypsin, L-glutamine, penicillin/streptomycin, D-glucose, 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), dimethyl sulfoxide, Triton X-100, RU486, HEPES, NaCl, trigonelline, and quercetin were purchased from Sigma. B-27 supplement was obtained from Thermo Fisher Scientific. Xanthohumol (purity 65–85%) was provided by Hopsteiner.
7
Nrf2 Modulation in α-LA Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
α-LA (R- α-LA) and a Nrf2 inhibitor, trigonelline, were purchased from Sigma-Aldrich (St. Louis, MO, USA) and were dissolved in 0.5 M ethanol and distilled water, respectively. A HO-1 inhibitor protoporphyrin (ZnPP) was purchased from Santa Cruz Biotechnology (sc-2000329, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and was dissolved in dimethyl sulfoxide (DMSO).
8
Evaluating Cell Apoptosis and Cytotoxicity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was evaluated by Annexin V/propidium iodide double staining assay. The cells were incubated at room temperature for 20 min in the dark and analyzed by flow cytometry. The number of each type of cells was expressed as percentages of the number of total stained cells.
LDH leakage assay was evaluated after collecting the culture medium and the cells by the Pierce LDH cytotoxicity assay kit. The cells were first sonicated to ensure the cell membrane broke down to release the total amount of LDH; subsequently, centrifugation (1,000 × g for 15 min) to clear up the cell sample was undertaken. LDH leakage was estimated from the ratio between the LDH activity in the culture medium and that of the whole cell content.
The requirement of UPR and Nrf2 pathways for promoting adaptation and survival to I-R was assessed using the following pharmacologic tools, at a concentration ranging from 1 to 3 µM: GSK compound 39 (Merck Millipore, Darmstadt, Germany), 4µ8c (Sigma Aldrich, Milan, Italy) and trigonelline (Sigma Aldrich, Milan, Italy) respectively for PERK, IRE1, and Nrf2 inhibition.
LDH leakage assay was evaluated after collecting the culture medium and the cells by the Pierce LDH cytotoxicity assay kit. The cells were first sonicated to ensure the cell membrane broke down to release the total amount of LDH; subsequently, centrifugation (1,000 × g for 15 min) to clear up the cell sample was undertaken. LDH leakage was estimated from the ratio between the LDH activity in the culture medium and that of the whole cell content.
The requirement of UPR and Nrf2 pathways for promoting adaptation and survival to I-R was assessed using the following pharmacologic tools, at a concentration ranging from 1 to 3 µM: GSK compound 39 (Merck Millipore, Darmstadt, Germany), 4µ8c (Sigma Aldrich, Milan, Italy) and trigonelline (Sigma Aldrich, Milan, Italy) respectively for PERK, IRE1, and Nrf2 inhibition.
9
Analytical Standards for Chlorogenic Acids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analytical standards of chlorogenic acid (5-CQA, CAS: 327-97-9, 5-O-caffeoylquinic acid), neochlorogenic acid (3-CQA, CAS: 906-33-2, 3-O-caffeoylquinic acid), caffeine (CAS: 58-08-2), and trigonelline (CAS: 535-83-1) were purchased from Sigma-Aldrich, Fluka, ChromaDex. All standards used were of analytical grade (≥ 99% purity). The 5-CQA standard was used to quantify both 5-CQA and 4-CQA. Mobile phase was prepared by diluting formic acid (CAS: 64-18-6, Fisher Scientific, LC–MS purity) with ultrapure water (Direct-Q system, resistivity below 18 MΩ cm) to a concentration of 0.1% (w/w) and the pH was adjusted to 2.4. The solution was filtered under a vacuum system through a 0.45 µm filter. The second part of the mobile phase was methanol (CAS: 67-56-1, POCH, HPLC grade). Stock solutions of 1000 mg/l were prepared by diluting each analysed compound in HPLC grade methanol. These stock solutions were diluted in methanol to reach intermediate concentrations. All solutions were stored in the refrigerator at 4 °C.
10
Analytical Standards for Antioxidant Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phenolic standards (gallic acid, cinnamic acid, chlorogenic acid, caffeic acid, coumaric acid, ferulic acid, 2,4-dihydroxybenzoic acid), quercetin, caffein, trigonelline, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and Folin–Ciocalteu’s reagent were purchased from Sigma–Aldrich (St. Louis, MO, USA). Chemicals used for the physicochemical analysis were purchased from Merck (Darmstadt, Germany). All chemicals used were analytical grade unless specified. All samples were stored in sealed glass jars and at room temperature (20–30 °C) until analysis
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!