KMID : 1138520180210010018
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Journal of Pharmacopuncture 2018 Volume.21 No. 1 p.18 ~ p.25
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Preventive effects of crocin on neuronal damages induced by D-galactose through AGEs and oxidative stress in human neuroblastoma cells (SH-SY5Y)
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Heidari Somaye
Mehri Soghra Shariaty Vahidesadat Hosseinzadeh Hossein
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Abstract
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Objective: D-galactose (D-gal) is well-known agent to induce aging process. In the present study, we selected crocin, the main constituent of Crocus sativus L. (saffron), against D-gal- induced cytotoxicity in human neuroblastoma SH-SY5Y cells.
Methods: Pretreated cells with crocin (25-500 ¥ìM, 24 h) were exposed to D-gal (25?400 mM, 48 h). The MTT assay was used for determination cell viability. Dichlorofluorescin diacetate assay (DCF-DA) and senescence associated ¥â-galactosidase staining assay (SA- ¥â-gal) were used to evaluate the generation of reactive oxygen species and beta-galactosidase as an aging marker, respectively. Also advanced glycation end products (AGEs) expression which is known as the main mechanism of age-related diseases was measured by western blot analysis.
Results: The findings of our study showed that treatment of cells with D-gal (25-400 mM) for 48h decreased cell viability concentration dependency. Reactive oxygen species (ROS) levels which are known as main factors in age-related diseases increased from 100 ¡¾ 8% in control group to 132 ¡¾ 22% in D-gal (200 mM) treated cells for 48h. The cytotoxic effects of D-gal decreased with 24h crocin pretreatment of cells. The cell viability at concentrations of 100 ¥ìM, 200 ¥ìM and 500 ¥ìM increased and ROS production decreased at concentrations of 200 and 500 ¥ìM to 111.5 ¡¾ 6% and 108 ¡¾ 5%, respectively. Also lysosomal biomarker of aging and carboxymethyl lysine (CML) expression as an AGE protein, significantly increased in D-gal 200 mM group after 48h incubation compare to control group. Pretreatment of SHSY-5Y cells with crocin (500 ¥ìM) before adding D-gal significantly reduced aging marker and CML formation.
Conclusion: Treatment of SH-SY5Y cells with crocin before adding of D-gal restored aging effects of D-gal concentration dependency. These findings indicate that crocin has potent anti- aging effects through inhibition of AGEs and ROS production.
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KEYWORD
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D-galactose, Crocin, Saffron, Aging, Reactive oxygen species, Advanced glycation end products
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