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Preventive effects of coixol, an active compound of adlay seed, in NGF-differentiated PC12 cells against beta-amyloid25-35-induced neurotoxicity Cover

Preventive effects of coixol, an active compound of adlay seed, in NGF-differentiated PC12 cells against beta-amyloid25-35-induced neurotoxicity

Asian Biomedicine
Volume 18 (2024): Issue 5 (October 2024)
By: Jan-Yow Chen,  Chien-Yu Li,  Mei-Chin Mong and  Mei-Chin Yin  
Open Access
|Oct 2024

Figures & Tables

Figure 1.

Structure of coixol (A) and adlay seeds (B).
Structure of coixol (A) and adlay seeds (B).

Figure 2.

Effects of coixol alone upon cell viability. NGF-differentiated PC12 cells were treated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, 2 μM, 4 μM, 8 μM, 16 μM, or 32 μM for 48 h. Normal group had no coixol. Data are expressed as mean ± SD (n = 8). †: P < 0.05 compared the normal; ‡: P < 0.05 compared coixol 8; §: P < 0.05 compared coixol 16; #: P < 0.05 compared coixol 32. NGF, nerve growth factor; SD, standard deviation.
Effects of coixol alone upon cell viability. NGF-differentiated PC12 cells were treated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, 2 μM, 4 μM, 8 μM, 16 μM, or 32 μM for 48 h. Normal group had no coixol. Data are expressed as mean ± SD (n = 8). †: P < 0.05 compared the normal; ‡: P < 0.05 compared coixol 8; §: P < 0.05 compared coixol 16; #: P < 0.05 compared coixol 32. NGF, nerve growth factor; SD, standard deviation.

Figure 3.

Effects of coixol on cell viability, Bcl-2 mRNA expression, and Bax mRNA expression. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. NGF, nerve growth factor; SD, standard deviation.
Effects of coixol on cell viability, Bcl-2 mRNA expression, and Bax mRNA expression. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. NGF, nerve growth factor; SD, standard deviation.

Figure 4.

Effects of coixol on intracellular Ca2+ level. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Intracellular Ca2+ level was measured at 0 h, 12 h, and 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. NGF, nerve growth factor; SD, standard deviation.
Effects of coixol on intracellular Ca2+ level. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Intracellular Ca2+ level was measured at 0 h, 12 h, and 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. NGF, nerve growth factor; SD, standard deviation.

Figure 5.

Effects of coixol on NF-κB binding activity. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. NF-κB, nuclear factor kappa B; NGF, nerve growth factor; SD, standard deviation.
Effects of coixol on NF-κB binding activity. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. NF-κB, nuclear factor kappa B; NGF, nerve growth factor; SD, standard deviation.

Figure 6.

Effects of coixol on protein expression of phosphorylated NF-κB p65, phosphorylated p38, iNOS, RAGE, and Nrf2. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. iNOS, inducible nitric oxide synthase; NF-κB, nuclear factor kappa B; NGF, nerve growth factor; RAGE, receptor of advanced glycation end product; SD, standard deviation.
Effects of coixol on protein expression of phosphorylated NF-κB p65, phosphorylated p38, iNOS, RAGE, and Nrf2. NGF-differentiated PC12 cells were pretreated with coixol at 0.125 μM, 0.25 μM, 0.5 μM, 1 μM, or 2 μM for 48 h, followed by Abeta25-35 exposure for 24 h. Normal groups had no coixol or Abeta25-35. Abeta groups had no coixol, but with Abeta25-35. Data are expressed as mean ± SD (n = 8). a: P < 0.05 compared the normal; b: P < 0.05 compared Abeta; c: P < 0.05 compared coixol 0.125 + Abeta; d: P < 0.05 compared coixol 0.25 + Abeta; e: P < 0.05 compared coixol 0.5 + Abeta; and f: P < 0.05 compared coixol 1 + Abeta. iNOS, inducible nitric oxide synthase; NF-κB, nuclear factor kappa B; NGF, nerve growth factor; RAGE, receptor of advanced glycation end product; SD, standard deviation.

Effects of coixol on the levels or activities of ROS, 8-OHdG, GSH, GPX, GR, and catalase

ROS RFU/mg protein8-OHdG ng/mg proteinGSH ng/mg proteinGPX U/mg proteinGR U/mg proteinCatalase U/mg protein
Normal0.13 ± 0.040.43 ± 0.0688 ± 572.1 ± 1.762.8 ± 1.170.4 ± 1.9
Abeta3.11 ± 0.21*2.72 ± 0.10*26 ± 2*30.2 ± 0.9*27.6 ± 0.7*24.1 ± 1.0*
Coixol, 0.125 + Abeta2.72 ± 0.14*2.51 ± 0.15*29 ± 4*32.5 ± 1.2*29.0 ± 1.2*30.5 ± 0.8*,**
Coixol, 0.25 + Abeta2.15 ± 0.08*,**,***2.02 ± 0.07*,**,***36 ± 3*,**,***43.1 ± 1.0*,**,***37.1 ± 1.0*,**,***39.3 ± 1.2*,**,***
Coixol, 0.5 + Abeta1.64 ± 0.15*,**,***,#1.73 ± 0.11*,**,***,#49 ± 5*,**,***,#52.4 ± 1.3*,**,***,#45.3 ± 0.9*,**,***,#48.4 ± 1.5*,**,***,#
Coixol, 1 + Abeta1.03 ± 0.17*,**,***,#,##1.36 ± 0.08*,**,***,#,##60 ± 4*,**,***,#,##55.0 ± 2.1*,**,***,#53.2 ± 1.4*,**,***,#,##55.9 ± 1.1*,**,***,#,##
Coixol, 2 + Abeta0.87 ± 0.09*,**,***,#,##1.13 ± 0.13*,**,***,#,##71 ± 3*,**,***,#,##,###62.7 ± 1.4*,**,***,#55.3 ± 0.8*,**,***,#,##60.3 ± 0.9*,**,***,#,##

Effects (% of normal groups) of coixol on MMP, Na+-K+ ATPase, and caspase-3 activities

MMPNa+-K+ ATPaseCaspase-3
Normal100100100
Abeta25 ± 5*30 ± 3*337 ± 19*
Coixol, 0.125 + Abeta29 ± 3*39 ± 2*,**315 ± 23*
Coixol, 0.25 + Abeta38 ± 4*,**,***44 ± 5*,**276 ± 11*,**,***
Coixol, 0.5 + Abeta49 ± 3*,**,***,#56 ± 2*,**,***,#230 ± 13*,**,***,#
Coixol, 1 + Abeta60 ± 5*,**,***,#,##68 ± 3*,**,***,#,##183 ± 8*,**,***,#,##
Coixol, 2 + Abeta71 ± 6*,**,***,#,##,###74 ± 5*,**,***,#,##172 ± 12*,**,***,#,##

Effects (pg/mg protein) of coixol on the levels of TNF-α, IL-1β, IL-6, and PGE2

TNF-αIL-1βIL-6PGE2
Normal12 ± 37 ± 29 ± 3109 ± 8
Abeta261 ± 14*199 ± 13*204 ± 9*442 ± 23*
Coixol, 0.125 + Abeta229 ± 10*,**162 ± 11*,**170 ± 10*,**386 ± 17*,**
Coixol, 0.25 + Abeta185 ± 9*,**,***120 ± 8*,**,***131 ± 8*,**,***319 ± 12*,**,***
Coixol, 0.5 + Abeta136 ± 12*,**,***,#84 ± 10*,**,***,#97 ± 6*,**,***,#257 ± 20*,**,***,#
Coixol, 1 + Abeta99 ± 7*,**,***,#,##61 ± 6*,**,***,#,##72 ± 5*,**,***,#,##192 ± 8*,**,***,#,##
Coixol, 2 + Abeta90 ± 5*,**,***,#,##52 ± 4*,**,***,#,##68 ± 4*,**,***,#,##177 ± 9*,**,***,#,##
DOI: https://doi.org/10.2478/abm-2024-0030 | Journal eISSN: 1875-855X | Journal ISSN: 1905-7415
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Language: English
Page range: 224 - 235
Published on: Oct 31, 2024
Published by: Chulalongkorn University
In partnership with: Paradigm Publishing Services
Publication frequency: 6 issues per year
Keywords:
Abeta,
Ca homeostasis,
coixol,
inflammatory stress,
NGF-differentiated PC12 cells,
oxidative stress
Related subjects:
Medicine,
Assistive professions, nursing,
Basic medical science,
Basic medical science, other,
Clinical medicine,
Clinical medicine, other

© 2024 Jan-Yow Chen, Chien-Yu Li, Mei-Chin Mong, Mei-Chin Yin, published by Chulalongkorn University
This work is licensed under the Creative Commons Attribution 4.0 License.

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