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

Jan-Yow Chen; Chien-Yu Li; Mei-Chin Mong; Mei-Chin Yin

doi:10.2478/abm-2024-0030

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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

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

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

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 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 Abeta_25-35 exposure for 24 h. Normal groups had no coixol or Abeta_25-35. Abeta groups had no coixol, but with Abeta_25-35. 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 Abeta_25-35 exposure for 24 h. Intracellular Ca²⁺ level was measured at 0 h, 12 h, and 24 h. Normal groups had no coixol or Abeta_25-35. Abeta groups had no coixol, but with Abeta_25-35. 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.

Table 1.

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

	MMP	Na⁺-K⁺ ATPase	Caspase-3
Normal	100	100	100
Abeta	25 ± 5^*	30 ± 3^*	337 ± 19^*
Coixol, 0.125 + Abeta	29 ± 3^*	39 ± 2^^,^*	315 ± 23^*
Coixol, 0.25 + Abeta	38 ± 4^^,^^,^**	44 ± 5^^,^*	276 ± 11^^,^^,^**
Coixol, 0.5 + Abeta	49 ± 3^^,^^,^**^,^#	56 ± 2^^,^^,^**^,^#	230 ± 13^^,^^,^**^,^#
Coixol, 1 + Abeta	60 ± 5^^,^^,^**^,^#^,^##	68 ± 3^^,^^,^**^,^#^,^##	183 ± 8^^,^^,^**^,^#^,^##
Coixol, 2 + Abeta	71 ± 6^^,^^,^**^,^#^,^##^,^###	74 ± 5^^,^^,^**^,^#^,^##	172 ± 12^^,^^,^**^,^#^,^##

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 Abeta_25-35 exposure for 24 h. Normal groups had no coixol or Abeta_25-35. Abeta groups had no coixol, but with Abeta_25-35. Data are expressed as mean ± SD (n = 8).

* P < 0.05 compared the normal.

** P < 0.05 compared Abeta.

*** P < 0.05 compared coixol 0.125 + Abeta.

# P < 0.05 compared coixol 0.25 + Abeta.

## P < 0.05 compared coixol 0.5 + Abeta.

### P < 0.05 compared coixol 1 + Abeta.

MMP, mitochondrial membrane potential; 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 Abeta_25-35 exposure for 24 h. Normal groups had no coixol or Abeta_25-35. Abeta groups had no coixol, but with Abeta_25-35. 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.

Table 2.

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

	ROS RFU/mg protein	8-OHdG ng/mg protein	GSH ng/mg protein	GPX U/mg protein	GR U/mg protein	Catalase U/mg protein
Normal	0.13 ± 0.04	0.43 ± 0.06	88 ± 5	72.1 ± 1.7	62.8 ± 1.1	70.4 ± 1.9
Abeta	3.11 ± 0.21^*	2.72 ± 0.10^*	26 ± 2^*	30.2 ± 0.9^*	27.6 ± 0.7^*	24.1 ± 1.0^*
Coixol, 0.125 + Abeta	2.72 ± 0.14^*	2.51 ± 0.15^*	29 ± 4^*	32.5 ± 1.2^*	29.0 ± 1.2^*	30.5 ± 0.8^^,^*
Coixol, 0.25 + Abeta	2.15 ± 0.08^^,^^,^**	2.02 ± 0.07^^,^^,^**	36 ± 3^^,^^,^**	43.1 ± 1.0^^,^^,^**	37.1 ± 1.0^^,^^,^**	39.3 ± 1.2^^,^^,^**
Coixol, 0.5 + Abeta	1.64 ± 0.15^^,^^,^**^,^#	1.73 ± 0.11^^,^^,^**^,^#	49 ± 5^^,^^,^**^,^#	52.4 ± 1.3^^,^^,^**^,^#	45.3 ± 0.9^^,^^,^**^,^#	48.4 ± 1.5^^,^^,^**^,^#
Coixol, 1 + Abeta	1.03 ± 0.17^^,^^,^**^,^#^,^##	1.36 ± 0.08^^,^^,^**^,^#^,^##	60 ± 4^^,^^,^**^,^#^,^##	55.0 ± 2.1^^,^^,^**^,^#	53.2 ± 1.4^^,^^,^**^,^#^,^##	55.9 ± 1.1^^,^^,^**^,^#^,^##
Coixol, 2 + Abeta	0.87 ± 0.09^^,^^,^**^,^#^,^##	1.13 ± 0.13^^,^^,^**^,^#^,^##	71 ± 3^^,^^,^**^,^#^,^##^,^###	62.7 ± 1.4^^,^^,^**^,^#	55.3 ± 0.8^^,^^,^**^,^#^,^##	60.3 ± 0.9^^,^^,^**^,^#^,^##

* P < 0.05 compared the normal.

** P < 0.05 compared Abeta.

*** P < 0.05 compared coixol 0.125 + Abeta.

# P < 0.05 compared coixol 0.25 + Abeta.

## P < 0.05 compared coixol 0.5 + Abeta.

### P < 0.05 compared coixol 1 + Abeta.

GPX, glutathione peroxidase; GR, glutathione reductase; GSH, glutathione; NGF, nerve growth factor; RFU, relative fluorescence unit; ROS, reactive oxygen species; 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 Abeta_25-35 exposure for 24 h. Normal groups had no coixol or Abeta_25-35. Abeta groups had no coixol, but with Abeta_25-35. 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.

Table 3.

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

	TNF-α	IL-1β	IL-6	PGE₂
Normal	12 ± 3	7 ± 2	9 ± 3	109 ± 8
Abeta	261 ± 14^*	199 ± 13^*	204 ± 9^*	442 ± 23^*
Coixol, 0.125 + Abeta	229 ± 10^^,^*	162 ± 11^^,^*	170 ± 10^^,^*	386 ± 17^^,^*
Coixol, 0.25 + Abeta	185 ± 9^^,^^,^**	120 ± 8^^,^^,^**	131 ± 8^^,^^,^**	319 ± 12^^,^^,^**
Coixol, 0.5 + Abeta	136 ± 12^^,^^,^**^,^#	84 ± 10^^,^^,^**^,^#	97 ± 6^^,^^,^**^,^#	257 ± 20^^,^^,^**^,^#
Coixol, 1 + Abeta	99 ± 7^^,^^,^**^,^#^,^##	61 ± 6^^,^^,^**^,^#^,^##	72 ± 5^^,^^,^**^,^#^,^##	192 ± 8^^,^^,^**^,^#^,^##
Coixol, 2 + Abeta	90 ± 5^^,^^,^**^,^#^,^##	52 ± 4^^,^^,^**^,^#^,^##	68 ± 4^^,^^,^**^,^#^,^##	177 ± 9^^,^^,^**^,^#^,^##

* P < 0.05 compared the normal.

** P < 0.05 compared Abeta.

*** P < 0.05 compared coixol 0.125 + Abeta.

# P < 0.05 compared coixol 0.25 + Abeta.

## P < 0.05 compared coixol 0.5 + Abeta.

### P < 0.05 compared coixol 1 + Abeta.

IL-1β, interleukin-1 beta; IL-6, interleukin-6; NGF, nerve growth factor; PGE₂, prostaglandin E₂; SD, standard deviation; TNF-α, tumor necrosis factor-α.

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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

Keywords:

Abeta_25-35,

Ca²⁺ 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.

Volume 18 (2024): Issue 5 (October 2024)

	TNF-α	IL-1β	IL-6	PGE₂
Normal	12 ± 3	7 ± 2	9 ± 3	109 ± 8
Abeta	261 ± 14^*	199 ± 13^*	204 ± 9^*	442 ± 23^*
Coixol, 0.125 + Abeta	229 ± 10^^,^*	162 ± 11^^,^*	170 ± 10^^,^*	386 ± 17^^,^*
Coixol, 0.25 + Abeta	185 ± 9^^,^^,^**	120 ± 8^^,^^,^**	131 ± 8^^,^^,^**	319 ± 12^^,^^,^**
Coixol, 0.5 + Abeta	136 ± 12^^,^^,^**^,^#	84 ± 10^^,^^,^**^,^#	97 ± 6^^,^^,^**^,^#	257 ± 20^^,^^,^**^,^#
Coixol, 1 + Abeta	99 ± 7^^,^^,^**^,^#^,^##	61 ± 6^^,^^,^**^,^#^,^##	72 ± 5^^,^^,^**^,^#^,^##	192 ± 8^^,^^,^**^,^#^,^##
Coixol, 2 + Abeta	90 ± 5^^,^^,^**^,^#^,^##	52 ± 4^^,^^,^**^,^#^,^##	68 ± 4^^,^^,^**^,^#^,^##	177 ± 9^^,^^,^**^,^#^,^##

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

Figures & Tables

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Table 1.

Figure 5.

Table 2.

Figure 6.

Table 3.

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