Background
Transcrocetin is a natural apocarotenoid isolated from C. sativus and G. jasminoides that has antioxidant, antiproliferative, anti-inflammatory, cardioprotective, and antinociceptive properties.1,2,3,4,5,6 It scavenges 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (IC50 = 17.8 μg/ml) and inhibits growth of MKN28 stomach, MCF-7 breast, and Caco-2 colon cancer cell lines (IC50s = 53, 63, and 103 μM, respectively).2,3 Transcrocetin (20 μM) protects primary rat microglial cells from LPS-induced death and decreases LPS-induced production of intracellular reactive oxygen species (ROS), TNF-α, IL-1β, and NF-κB.4 Transcrocetin (100 mg/kg) increases the level of glutathione (GSH), catalase (CAT), creatine kinase (CK), and lactate dehydrogenase (LDH) in cardiac tissue in a rat model of myocardial infarction induced by isoproterenol .5 It also increases the pressure threshold and latency to withdrawal in response to mechanical and thermal stimuli, respectively, indicating a decrease in allodynia in a mouse model of spared nerve injury when administered intrathecally at a dose of 30 mg/kg.6
1.Pfister, S.L., Meyer, P., Steck, A., et al.Isolation and structure elucidation of carotenoid?glycosyl esters in gardenia fruits (Gardenia jasminoides Ellis) and saffron (Crocus sativus Linne)J. Agric. Food Chem.44(9)2612-2615(1996)
2.Kanakis, C.D., Tarantilis, P.A., Tajmir-Riahi, H.A., et al.Crocetin, dimethylcrocetin, and safranal bind human serum albumin: Stability and antioxidative propertiesJ. Agric. Food Chem.55(3)970-977(2007)
3.Oliveira, H., Cai, X., Zhang, Q., et al.Gastrointestinal absorption, antiproliferative and anti-inflammatory effect of the major carotenoids of Gardenia jasminoides Ellis on cancer cellsFood Funct.8(4)1672-1679(2017)
4.Nam, K.N., Park, Y.-M., Jung, H.-J., et al.Anti-inflammatory effects of crocin and crocetin in rat brain microglial cellsEur. J. Pharmacol.648(1-3)110-116(2010)
5.Zhang, W., Li, Y., and Ge, Z.Cardiaprotective effect of crocetin by attenuating apoptosis in isoproterenol induced myocardial infarction rat modelBiomed. Pharmacother.93376-382(2017)
6.Wang, X., Zhang, G., Qiao, Y., et al.Crocetin attenuates spared nerve injury-induced neuropathic pain in miceJ. Pharmacol. Sci.135(4)141-147(2017)
Protocol
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Cell experiment:
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Cytotoxicity of test compounds is determined by MTT assay using Caco-2 cells in 96 well plates at a density of 20.000 cells per well in 200 μl FBS-free medium, grown for 96 h and followed by 24 h contact time with the test compounds (100 μL of serum-free media containing SE 0.5, 1, and 2 mg/mL; trans-crocin-1 250, 500, and 1000 μM; Transcrocetin 10, 40, 80, and 160 μM) and incubation at 37°C/5% CO2. The incubation solutions are aspirated, each well is washed twice with 150 μL of PBS and 50 μL of MTT solution are added (2.5 mg/mL in PBS). Supernatants are discarded and the formed formazan is dissolved in 50 μL of DMSO. The absorption of the resulting solution is determined at λ=492 nm against reference wavelength λ=690 nm[1].
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参考文献:
[1]. Lautenschl?ger M, et al. Intestinal formation of trans-Crocetin from saffron extract (Crocus sativus L.) and in vitro permeation through intestinal and blood brain barrier. Phytomedicine. 2015 Jan 15;22(1):36-44.
[2]. José Bagur M, et al. Saffron: An Old Medicinal Plant and a Potential Novel Functional Food. Molecules. 2017 Dec 23;23(1). pii: E30.
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