Table 2.

The results of in vitro experiments, in which various protective agents were used against acrylamide toxicity in various cell lines

Article number of references	Method	Acrylamide’s effect and dose	Protective agent’s effect and dose
1-Hong et al. (16), 2019	-ROS formation	↑	all ↓
	-SOD	↓	15 and 30 µM↑
	-GPx1	↓	7.5, 15 and 30 µM↑
	-DNA fragmentation	↑	7.5, 15 and 30 µM↑
	Western Blot
	-pJNK/JNK	↑	all↓
	-pERK/ERK	↓	all↑
	-p-p38/p38	↑	all↓
2-Yıldızbayrak and Erkan (17), 2019	-Cell viability	10, 100, 1000 µM ↓	↑
	-Apoptosis and necrosis	10, 100, 1000 µM ↑	↓
	-OH- production	10, 100, 1000 µM ↑	100, 1000 µM ↓
	-H2O2 production	1000 µM ↑	NS
	-MDA	10, 100, 1000 µM ↑	100, 1000 µM ↓
	-SOD	10, 100, 1000 µM ↓	NS
	-CAT	100, 1000 µM ↓	100, 1000 µM ↑
	-GPx1	10, 100, 1000 µM ↓	all doses ↑
	Western Blot
	-pJNK/JNK	10, 100, 1000 µM ↑	10, 100, 1000 µM↓
	-pERK/ERK	10, 100, 1000 µM ↑	100, 1000 µM↓
	-p-p38/p38	10, 100, 1000 µM ↑	1000 µM↓
3-Azari et al. (18), 2019	-Cell viability	↓	200 µM ↑
	-ROS formation	↑	all ↓
	-MDA	↑	100 and 200 µM ↓
	-GSH	↓	100 and 200 µM ↑
	-Protein carbonyl	↑	100 and 200 µM ↓
4-Pan et al. (19), 2018	-Cell viability	↓	↑
	-ROS formation	↑	↓
	-MDA	↑	↓
	-TNF-α	↑	↓
	-IL-6	NS	NS
	Western Blot
	-Nrf-2	↑	↑
	-NFκBp65	NS↑	↓
	Note: *non-significant increase
5-Jiang et al. (20), 2018	-LDH	↑	all doses ↓
	-ROS	4-hour AA↑	all doses ↓
	- Annexin V/PI (apoptosis)	24-hour AA ↑	all doses ↓
	-Disrupted MMP	4-hour AA ↑	all doses ↓
	-SOD	16-hour AA ↓	40 µg/mL doses ↑
	-GPx1	16-hour AA ↓	all doses ↑
	-MDA	16-hour AA ↑	all doses ↓
	Western Blot (4-hour AA)
	-Bax	↑	all doses ↓
	-Bcl-2	↓	all doses ↑
	-Casp3	↑	all doses ↓
	-Cleaved casp3	↑	all doses ↓
	-Casp9	↑	all doses ↓
	-Cleaved casp9	↑	all doses ↓
	-Cyt c	↑	all doses ↓
6-Esmaeelpanah et al. (21), 2018	-Cell viability	↓	both agents ↑
	-GSH	↓	both agents ↑
	-MDA	↑	10 and 20 µM for ECG
			↓, 20 µM for EGCG↓
7-Albalawi et al. (22), 2017	-Cell viability	↓	↑
	-TUNEL (apoptotic cells)	↑	↓
	-ROS	↑	↓
	-SOD	↓	↑
	-MDA	↑	↓
	-GSH	↓	↑
	-CAT	↓	↑
	Real-time PCR
	-Casp 3	↑	↓
	-Casp 9	↑	↓
	-Nrf2	↓	↑
	-GPx1	↓	↑
	-SOD1	↓	↑
	-CAT	↓	↑
	-NQO1	0.7 M NS, 1 mM↓	↑
	-GCLM	NS	↑
	Western Blot
	-Nrf2	↓	↑
8-Li et al. (23), 2018	-Cell viability	↓	all AE doses ↑
	-SOD	↓	all AE doses ↑
	-CAT	↓	all AE doses ↑
	-MDA	↑	all AE doses ↓
9-Song et al. (24), 2013	-Cell viability	↓	↑
	-LDH	↑	↓
	-ROS	↑	25, 50, 100 µM ↓
	-GSH	↓	10, 25, 50 µM ↑
	-GPx1	↑	all C3G doses ↓
	-GST	↑	all C3G doses ↓
	Western Blot
	-GPx1	↓	all C3G doses ↑
	-GSTP1	↓	all C3G doses ↑
	- λ-GCS	↓	all C3G doses ↑
	-CYP2E1	↑	50 and 100 µM ↓
10-Mehri et al. (25), 2012	-Cell viability	↓	all CRO doses ↑
	-DNA fragmentation	↑	20 and 50 µM ↓
	-Annexin V/PI (apoptosis)	↑	all CRO doses ↓
	-Annexin V/PI (necrosis)	↑	20 and 50 µM ↓
	- ROS	↑	all CRO doses ↓
	Western Blot
	-Bax	↑	↓
	-Bcl-2	↓	NS (Western bands
			with similar with acrylamide group)
	-Bax/Bcl-2	↑	↓
11-Song et al. (26), 2017	-Cell viability	↓	↑
	AO/EB	NQ	NQ
	-PGE2	↑	↓
	-TNF-α	↑	↓
	-IL-1β	↑	↓
	-NO	↑	↓
	-GSH	↓	↑
	-H2O2	↑	↓
	Western Blot	↑	↓*
	-p-ERK/ERK	↑	↓*
	-pp38/p38	↑	↓*
	-p-JNK/JNK	↑	↓*
	-p-IκB/IκB	↑	↓*
	-NFκB-nuclear	↑	↓*
	-Bax	↓	↑*
	-Bcl-2	↑	↓*
	-Cyt-c	↑	↓*
	-Cleaved-Casp3	↓	↑*
	-pAKT	↑	↓*
	-p-GSK-3β	↑	↓
	-Bax/Bcl-2	↑	↓*
	-MPO
	*note: The change was inferred from the density of Western bands based on observation because most western bands had not been quantified.
12-He et al. (27), 2017	-Cell viability	↓	-5, 10 and 20 µM ↑
	-Acetylcholinesterase activity	↓	-5 and 10 µM↑
	-Annexin V/PI	↑	-5 and 10 µM↓
	-MMP	↓	-5 and 10 µM↑
	- Intracellular Ca+2	↑	-5 and 10 µM↓
	-Cyt c	↑	-5 and 10 µM↓
	-Casp3	↑	-5 and 10 µM↓
	-ROS	↑	-5 and 10 µM↓
	-SOD	↓	-5 and 10 µM↑
	-MDA	↑	-5 and 10 µM↓
	-GSH	↓	-5 and 10 µM↑
	Real-Time PCR
	-Bax	↑	↓
	-Bcl-2	↓	NS
	-Bax/Bcl-2	↑	↓ (only 5 µM EGCG was used)
13-Chen et al. (28), 2014	-Cell viability	↓	all HP doses ↑
	-ROS	↑	all HP doses ↓
	-MMP (rhodamine123)	↓	all HP doses ↑
	-GSH (NDA)	↓	all HP doses ↑
14-Li et al. (29), 2017	-Cell viability	↓	-↑ 48 µg/ml for 1.5 and 3 hours SM –pre- treatments; 96 and 192 µg/ml ↑ for 1.5, 3 and 6 hours SM pre-treatments
	-ROS	↑	-48, 96 and 192 µg/ml↓
	-MDA	↑
	-GSH	↓	-all 5 SM doses↓
	Real-Time PCR		-48, 96 and 192 µg/ml↓
	-Total Nrf-2	NA	-24, 48, 96, 192 µg/ml↑
	-Gpx1	NA	-48, 96 and 192 µg/ml↓
	- GCLC	NA
	-GCLM	NA	-24, 48, 96, 192 µg/ml↑
	Western Blot
	-Total Nrf-2	NA	-all 5 SM doses↓
	-Nuclear Nrf2	↑
	-Cytoplasmic Nrf2	↓	-96 and 192 µg/ml ↑
	-Gpx1,	NA	-48, 96 and 192 µg/ml ↑
	-GCLC	NA
	-GCLM	NA	-all 5 SM doses↓
			-48, 96 and 192 µg/ml↑
			-48, 96 and 192 µg/ml↑
			-all 5 SM doses↓
15-Shi et al. (30), 2018	-Cell viability,	↓	-all CN/GCN doses↑
	-LDH	↑	-all CN/GCN doses ↓
	-TEER	↓	-↑25 µg/ml is the best
			for CN/GCN.
	-Paracellular permeability (destruction of TJs)	↑	-↓48 hours and 25 µg/ml are the best for CN/GCN.
	Real-Time PCR
	-ZO1, Claudin 1 and 3	↓	-CN/GCN digests for 24 /48 hours ↑
	-ZO2, Occludin		-CN digest for 24 /48 hours ↑. GCN digest for 24 hours ↑, for 48 hours NS.
	-Claudin 4	↓	-CN/GCN digests for 24/ 48 hours NS
	Western Blot
	-ZO1, Occludin and Claudin 1	↓	-CN/GCN for 24/48 hours↑
16-Chen et al. (31), 2013	Cell viability	↓	5 and 10 µg/mL↑
	ROS	↑	5 and 10 µg/mL↓
17-Rodriguez-Ramiro et al. (32), 2011	-Cell viability	↓	EC↑<PB2↑=CPE↑
	-ROS	↑	EC↓<PB2↓=CPE↓
	-GSH	↓	EC↑<PB2↑=CPE↑
	-Casp 3	↑	EC↓<PB2↓=CPE↓
	Western Blot
	-λ-GCS	↓	EC↑<PB2↑=CPE↑
	-λ-GST	↓	EC(NS)< PB2↑=CPE↑
	-pERK/ERK	↑	EC(NS) <PB2↓=CPE↓ EC(NS) <PB2↓=CPE↓
	-pJNK/JNK	↑
18-Rodriguez-Ramiro et al. (33), 2011	-Cell viability	↓	all HT doses ↑
	-Caspase 3 activity	↑	10, 20, 40 µM HT doses ↓
	-GSH (fluorometric)	↓	all HT doses NS
	-ROS	↑	all HT doses↓
	-GPx1	↑	10, 20, 40 µM HT doses ↓
	-GR activity	↑	all HT doses↓
	-pJNK/JNK	↑	all HT doses↓
19-Sumizawa and Igisu (34), 2009	-Cell viability	↓	10, 30, 60 and 100 µM CF doses ↑
	-Caspase 3	↑	↓
	-SubG1 cell population (cell -cycle analysis)	↑	↓
	-Acrylamide uptake	NA	60 and 120 µM CF doses ↑
	-GSH	↓	↑
20-Zhang et al. (35), 2009	-Cell viability	↓	all HT doses ↑
	-DNA tail length (Comet assay) -	↑	all HT doses ↓
	ROS	↑	all HT doses ↓
	-DNA damage (8-OHdG)	↑	all HT doses ↓
	-GSH	↓	25 and 50 µM↑
21- Cao et al. (36), 2008	-Cell viability	↑	↓
	-ROS	↑	↓
	-DNA tail length (Comet assay)	↑	↓
	-Micronucleus frequency	↑	↓

"↑", "↓" and "NS" indicate a significant increase, a significant decrease and non-significant change, respectively, when compared to control group in the column titled acrylamide’s effect and dose, and when compared to the acrylamide group in the column titled protective agent’s effect and dose. The first arrow direction (arrows in left-hand-side) shows the significant difference of acrylamide-induced cells when compared to control, the second arrow direction (arrows in right-hand-side) shows a significant difference of protective agent cells when compared to acrylamide-treated cells. In the Table and manuscript, only the data of the reviewed article that is related to the effect of the protective agents, and the only experiments that are related to in vitro cell line are used and presented. AA; Acrylamide, AE; Anthocyanin extract, AO/EB; Acridine orange/ Ethidium bromide, Bax; Bcl-2-associated X protein, C3G; Cyanidin-3-glucoside, Casp; Caspase, CAT; Catalase, CF; Carboxyfullerene, CN; Caseinate digest, CPE; Cocoa polyphenolic extract, CRO; Crocin, CYP2E1; Cytochrome P450 2E1, Cyt c; Cytochrome c, EC; Epicatechin, ECG; Epicatechin gallate, EGCG; Epigallocatechin gallate, ERK; Extracellular signal-regulated kinase, λ-GCS; Gamma-glutamylcysteine synthetase, GCLC; Glutamate-cysteine ligase catalytic subunit, GCLM; Glutamate-cysteine ligase modifier subunit, GCN; Glycated caseinate digest, GPx1; Glutathione peroxidase 1, GR; Glutathione reductase, GSH; Glutathione, GSK-3β; Glycogen synthase kinase 3 beta, GST; Glutathione S-transferase, GSTP1; Glutathione S-transferase P, H2O2; Hydrogen peroxide, HT; Hydroxytyrosol, IL-1β; Interleukin 1β, IL-6; Interleukin 6, JNK; c-Jun N-terminal kinase, LDH; Lactate dehydrogenase, MDA; Malondialdehyde, MMP; Mitochondrial membrane potential, MPO; Myeloperoxidase, NA; Not applicable, there is no given knowledge about it, NDA; Naphthalene-2, 3-dicarboxaldehyde, NFκB; Nuclear factor kappa B, NO; Nitric oxide, NQ; Not quantified data, NQO1; NAD(P)H quinone dehydrogenase 1, Nrf-2; Nuclear factor erythroid 2-related factor, PB2; Procyanidin B2, PGE2; Prostaglandin E2, PI; Propidium iodide, ROS; Reactive oxygen species, SM; Silymarin, SOD; Superoxide dismutase, TEER; Transepithelial/transendothelial electrical resistance, TJ; Tight junction, and TNF-α; Tumor necrosis factor α.