Effect of Melatonin on Protecting the Jejunum Mucosal Epithelial Cells from Oxidative Stress Damage

doi:10.13560/j.cnki.biotech.bull.1985.2023-0190

Abstract

Abstract:

The oxidative injury of intestinal mucosa is strongly associated with the development and intestinal diseases such as diarrhea for animals. Melatonin（N-acetyl-5-methoxy-tryptamine）, as an indoleamine neurohormone, has important role in the processes of antioxidant protection. To investigate the protective effect of melatonin on the oxidative injury to the mucosal epithelial cells, the primary mouse jejunum mucosal epithelial cells were isolated and exposed to the FeSO₄and H₂O₂ to establish the oxidative damage model. After adding different doses of melatonin to the epithelial cells, we observed the cells’ pathological changes and then detected the changes of oxidative products, antioxidant enzymes, and inflammatory-related cytokines. It is found that after treatment with FeSO₄and H₂O₂, the mucosal epithelial cells programmed severe injury with significant increase of malonaldehyde and down-regulation of antioxidant enzymes. When adding melatonin to epithelial cells, the significantly-decreased malonaldehyde and highly-expressed antioxidant enzymes were detected. Meanwhile, melatonin also significantly reduced the expression of pro-inflammatory factor interleukin-6, and increased the expression of chemotactic factor interleukin-8. Therefore, melatonin may alleviate the oxidative damage of mucosal epithelial cells of jejunum caused by the increase of free radicals, and reduce the inflammatory response induced by oxidative damage.

Key words: melatonin, oxidative stress, antioxidant, mucosal immunity

KANG Ling-yun, HAN Lu-lu, HAN De-ping, CHEN Jian-sheng, GAN Han-ling, XING Kai, MA You-ji, CUI Kai. Effect of Melatonin on Protecting the Jejunum Mucosal Epithelial Cells from Oxidative Stress Damage[J]. Biotechnology Bulletin, 2023, 39(9): 291-299.

Figures/Tables 7

References 39

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基因Gene	登录号GenBank ID	引物序列Primer sequence（5'-3'）	产物长度Product length/bp
TNF-α	NM_001024860.1	F: GAGAAGATTCGTGGGCTGAT	169
TNF-α	NM_001024860.1	R: CCTTGTTCTTGTCCTGTGGG	169
IL-1β	NM_001009465.2	F: TGATGGCTTATTACAGTGGCAA	183
IL-1β	NM_001009465.2	R: CCATGGCCACAACAACTGAC	183
IL6	NM_001009392.1	F: CCTTCGGTCCAGTTGCCTTCT	234
IL6	NM_001009392.1	R: CCAGTGCCTCTTTGCTGCTTTC	234
β-actin	NM_0010009784.3	F: GATTCCTATGTGGGCGACGA	229
β-actin	NM_0010009784.3	R: AGGTCTCAAACATGATCTGGGT	229

基因Gene	登录号GenBank ID	引物序列Primer sequence（5'-3'）	产物长度Product length/bp
TNF-α	NM_001024860.1	F: GAGAAGATTCGTGGGCTGAT	169
TNF-α	NM_001024860.1	R: CCTTGTTCTTGTCCTGTGGG	169
IL-1β	NM_001009465.2	F: TGATGGCTTATTACAGTGGCAA	183
IL-1β	NM_001009465.2	R: CCATGGCCACAACAACTGAC	183
IL6	NM_001009392.1	F: CCTTCGGTCCAGTTGCCTTCT	234
IL6	NM_001009392.1	R: CCAGTGCCTCTTTGCTGCTTTC	234
β-actin	NM_0010009784.3	F: GATTCCTATGTGGGCGACGA	229
β-actin	NM_0010009784.3	R: AGGTCTCAAACATGATCTGGGT	229

抗氧化因子 Antioxidant factor	不同处理组 Treatment groups							SEM	P
抗氧化因子 Antioxidant factor	CON	FeSO₄/H₂O₂	MT0.01	MT0.1	MT1.0	MT5.0	MT10	SEM	P
MDA	4.31^bc	5.21^a	4.37^bc	4.17^c	4.37^bc	4.57^b	4.57^b	0.099	<0.0001
LDH	3.74^a	3.88^a	3.14^b	3.44^ab	3.08^b	3.85^a	3.63^a	0.132	0.0011
SOD	1.94^ab	1.90^b	1.90^b	1.97^ab	1.94^ab	1.97^ab	2.00^a	0.022	0.068
GSH-Px	3.76^bc	3.60^c	3.96^ab	4.09^ab	4.25^a	4.12^ab	4.34^a	0.104	0.002
CAT	216.58^a	167.22^b	165.83^b	174.87^b	163.45^b	179.04^b	169.65^b	6.008	<0.01

抗氧化因子 Antioxidant factor	不同处理组 Treatment groups							SEM	P
抗氧化因子 Antioxidant factor	CON	FeSO₄/H₂O₂	MT0.01	MT0.1	MT1.0	MT5.0	MT10	SEM	P
MDA	4.31^bc	5.21^a	4.37^bc	4.17^c	4.37^bc	4.57^b	4.57^b	0.099	<0.0001
LDH	3.74^a	3.88^a	3.14^b	3.44^ab	3.08^b	3.85^a	3.63^a	0.132	0.0011
SOD	1.94^ab	1.90^b	1.90^b	1.97^ab	1.94^ab	1.97^ab	2.00^a	0.022	0.068
GSH-Px	3.76^bc	3.60^c	3.96^ab	4.09^ab	4.25^a	4.12^ab	4.34^a	0.104	0.002
CAT	216.58^a	167.22^b	165.83^b	174.87^b	163.45^b	179.04^b	169.65^b	6.008	<0.01