Page 5 - A. Aging

Protective Effect of Capsicum annuum L. (Paprika) Extract Fermented by L. plantarum
on Sodium Iodate-Induced Acute Retinal Degeneration in Mice
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Ha-Rim Kim , Sol Kim , Sang-Jun Kim , Sang-Wang Lee , Hong-Sig Sin , Seung-Il Jeong , Kang-Yeol Yu , Seon-Young Kim *
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1 Jeonju Agro-Biomaterials Institute, Jeonju, 54810, Republic of Korea
2 Chebigen Co. Ltd, Jeonju, 54853, Republic of Korea
BACKGROUND & AIM METHODS
Diseases of the outer retina, including age-related macular Paprika (Capsicum annuum L.) is widely used
degeneration (AMD), are major cause of the permanent visual damage. as a vegetable and food additive, commonly
The pathogenesis of AMD involves oxidative stress and damage of the called sweet pepper. Carotenoids have known
to have a role in maintaining health and
retinal pigment epithelium. Capsicum annuum L. (paprika) fruits have prevention of diseases by acting as an
been known as a source of vitamins, carotenoids, phenolic compounds, antioxidant, protecting cells and tissues
and metabolites with well-known antioxidant activity, which have positive Lactobacillus against damaging reactive oxygen species
effects on human health and protection against AMD and cataracts. In fermentation plantarum (ROS). Various carotenoids are likely to use
agents to prevent these diseases.
this study, we investigated whether treatment of fermented paprika
extract (FPE) using Lactobacillus (L.) plantarum could protect retinal LPS-induced in RAW264.7 macrophage
degeneration in mice. We developed a retinal damage model in
C57BL/6 mice via intraperitoneal injections using 30 mg/kg NaIO3. Nitric oxide assay
Decreases in deformation and thickness were observed in outer nuclear
layer 7 days after NaIO3 administration but were improved with FPE Western blot
treatment. These results show that NaIO3 can be varied to permanent ① MAPK signaling pathway
deficits in RPE and visual function. FPE treatment protected the
reduction of glutathione and superoxide dismutase levels. Additionally, ② NADPH signaling pathway
we evaluated the effect of FPE on lipopolysaccharide (LPS)-mediated
reactive oxygen species (ROS) production and its underlying Acute Retinal Degeneration in Mice
mechanisms in RAW 264.7 cells. LPS-induced nitric oxide and ROS
generation was markedly blocked by FPE treatment in RAW 264.7 cells. Retinal degeneration mice model Hematoxylin and eosin stain
Furthermore, we found that FPE regulated LPS-induced inducible nitric SOD activity assay
oxide synthase, cyclooxygenase-2, NAD(P)H oxidase signaling protein,
and mitogen activated kinase signaling protein expression in RAW Glutathione assay
264.7 cells. These findings suggest that FPE can be a potential Western blot
candidate for protecting the retinal degenerative diseases. ① MAPK signaling pathway
RESULTS
A Fig. 1. Effects of fermented paprika extract (FPE) A
on lipopolysaccharide (LPS)-induced Nitric oxide
(NO) and mitogen activated protein kinase (MAPK)
signaling in RAW264.7 cells.
A; NO production was determined in culture
supernatant using Griess reagent. B; LPS-induced
MAPK (ERK, JNK, and p38) phosphorylation in whole
cell lysates was determined by western blotting. Equal
amounts of protein were resolved by SDS-PAGE.
RAW 264.7 cells were pretreated with indicated
concentration (500 μg/mL) of fermented paprika B C
extract (FPE) for 1 h, and then followed by LPS (1
μg/mL) for 24h. C; Quantitative analysis of blots. N,
B
Normal; C, LPS control; YP, Yellow Paprika; FYP,
Fermented YP; OP, Orange Paprika; FOP, Fermented
OP. Data are shown as mean ± SD. ** p < 0.01, *** p
< 0.001 versus N group; ## p < 0.01, ### p < 0.001
versus C group.
A
D
C
B Fig.3. Effects of FPE on sodium iodate (SI)-induced acute retinal injury, oxidative
stress, phosphorylation of Akt and MAPK in mice.
A; Representative histopathologic changes by FRE administration in SI-treated eye tissues,
Fig. 2. Effects of FPE on LPS-induced stained with H&E. GCL, Ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear
intracellular reactive oxygen species (ROS) layer; RPE, retinal pigment epithelium. B; The levels of Superoxide dismutase (SOD) and
generation and inducible nitric oxide glutathione (GSH) in serum and eye tissue lysate were analyzed using SOD and GSH assay
(iNOS), cyclooxygenase (COX-2), and kits. C; The phosphorylation of Akt and MAPK; ERK, JNK, and p-38, in eye tissue lysates
NAD(P)H oxidase (NOX) activation. were determined by western blotting. Equal amounts of total protein were resolved by SDS-
A; The levels of protein of iNOS, COX-2, PAGE. D; Quantitative analysis of blots. N, Normal; C, SI control; YP, Yellow Paprika; FYP,
NOXs; p22, p67, MOX1, NOX2, and NOX4, Fermented-YP; OP, Orange Paprika; FYP, Fermented-OP. Data are shown as mean ± SD.
were determined by western blotting. Equal * p < 0.05, ** p < 0.01, *** p < 0.001 versus N group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus C
amounts of protein were resolved by SDS- group.
PAGE. B; Quantitative analysis of blots. N,
Normal; C, LPS control; YP, Yellow Paprika; CONCLUSION
FYP, Fermented YP; OP, Orange Paprika;
FOP, Fermented OP. Data are shown as Fermented paprika extract (FPE) using L. plantarum have shown
mean ± SD. *** p < 0.001 versus N group; # p protective effect of deformation in outer nuclear layer and anti-oxidant
< 0.05, ## p < 0.01 and ### p < 0.001 versus C
group. effect in acute retinal degeneration in mice. Furthermore, we found
that FPE regulated LPS-induced inducible nitric oxide synthase,
REFERENCES cyclooxygenase-2, NAD(P)H oxidase activation, and mitogen
activated kinases protein overexpression in RAW 264.7 cells. These
1. YAO, H., et al. Redox regulation of lung inflammation: role of NADPH oxidase and NF-κB
signalling. 2007. findings suggest that fermentation of paprika using L. plantarum can
enhance the potential for protecting the retinal degenerative diseases.
2. ZHANG, Jinglin, et al. Protection of retina by mini-αA in NaIO3-induced retinal pigment
epithelium degeneration mice. International Journal of Molecular Sciences, 2015, 16.1: 1644-
1656.
3. HE, Huijun, et al. Glycyrrhizin protects against sodium iodate‐induced RPE and retinal injury Contact information
though activation of AKT and Nrf2/HO‐1 pathway. Journal of cellular and molecular medicine,
2019, 23.5: 3495-3504. Correspondence: Seon02@jami.re.kr; Tel.: +82-63-711-1053