芦丁对脂多糖诱导的瘤胃上皮细胞屏障功能、炎症损伤的影响

doi:10.16431/j.cnki.1671-7236.2026.02.046

Abstract

Abstract:

Objective The purpose of this experiment was to investigate the effects of rutin on the barrier function and inflammatory injury of sheep rumen epithelial cells induced by lipopolysaccharide （LPS）. Method Sheep rumen epithelial cells were cultured in vitro. Different concentrations of LPS （0， 1， 5， 10， 50， and 100 μg/mL） were used to stimulate and establish a model of rumen epithelial cell injury. The optimal concentration and duration of LPS action were determined. The rumen epithelial cells were pre-treated with different concentrations of rutin （0， 25， 50， 100， and 150 μg/mL）， and the cell viability was detected by MTT assay. The relative proliferation rate of the cells was calculated to determine the appropriate concentration and duration of action of rutin. Based on the selected concentrations and durations of LPS and rutin， the cells were grouped and treated accordingly. They were respectively designated as control group （CON）， LPS group， rutin group （R）， and rutin intervention group （R+LPS）. The contents of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and IL-10 in the cell supernatant were determined by ELISA kit. The relative mRNA expression levels of IL-1β， IL-4， IL-6， IL-8， IL-10， IL-12， TNF-α， IFN-γ， CXCL8， CXCL9， Occludin， ZO-1， Claudin-1， and Claudin-4 were detected by Real-time quantitative PCR. Result Through experimental screening， it was ultimately determined that 1 μg/mL LPS， when applied for 9 hours， was the optimal condition for establishing the rumen epithelial cell injury model. A treatment of 100 µg/mL rutin for 8 hours was identified as the optimal condition for the pre-protective effect of rutin on rumen epithelial cells in vitro. Compared with CON group， the contents of IL-1β and TNF-α in the cell supernatant of LPS group were significantly increased （P<0.05）， while IL-10 content was significantly decreased （P<0.05）. Compared with LPS group， the contents of IL-1β and TNF-α in the cell supernatant of R+LPS group were significantly decreased （P<0.05）， while IL-10 content was significantly increased （P<0.05）. The Real-time quantitative PCR results showed that compared with CON group， the mRNA expression levels of IL-1β， IL-6， IL-12， TNF-α， IFN-γ， CXCL8， and CXCL9 in the rumen epithelial cells of LPS group were significantly increased （P<0.05）， the mRNA expression levels of IL-4， IL-10， Claudin-4， and ZO-1 were significantly decreased （P<0.05）. Compared with LPS group， the mRNA expression levels of IL-1β， IL-6， IL-12， TNF-α， INF-γ， CXCL8， and CXCL9 in the rumen epithelial cells of R+LPS group were significantly decreased （P<0.05）， the mRNA expression levels of IL-4， IL-10， ZO-1， and Claudin-4 were significantly increased （P<0.05）. Conclusion Under this experimental conditions， 100 µg/mL rutin， when applied for 8 hours， exhibited a significant protective effect on the barrier function and inflammatory injury of the rumen epithelial cells induced by LPS.

Key words: rumen epithelial cells; rutin; lipopolysaccharide; inflammatory factors; barrier function

CLC Number:

S816.7

HU Honglian, LI Lanzhu, XU Xiaohui, ZHAO Haixia, ZHAO Meng, SONG Liwen, WANG Jianjun, SUN Manji. Effects of Rutin on Barrier Function and Inflammatory Injury of Rumen Epithelial Cells Induced by Lipopdysaccharide[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(2): 1033-1042.

Figures/Tables 10

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References 23

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基因 Genes	引物序列 Primer sequences （5′→3′）	GenBank登录号 GenBank accession No.	产物长度 Product length/bp
IL-1β	F：AGTGATGGCTTGCTACAGTGATGAG R：CAGGTGTTGGGTGCAGCTCTTC	NM_001009465.2	85
IL-4	F：ACAAATTCCTGGGCGGACTTGAC R：CTCAGCGTACTTGTACTCGTCTTGG	NM_001009313.3	81
IL-6	F：ACGAGTGGGTAAAGAACGCAAAGG R：GCCGCAGCTACTTCATCCGAATAG	NM_001009392.1	99
IL-8	F：GCTGGCTGTTGCTCTCTTGGC R：GGGGTGGAAAGGTGTGGAATGTG	KC912524.1	127
IL-10	F：GGGTGTCTACAAAGCCATGAGTGAG R：AGGTTTATGTCGGGGAGTCTAGTCG	NM_001009327.1	143
IL-12	F：TCACAGACCAAACCTCAGCCAAAG R：CAGGACACAGATGCCCATTCACTC	XM_042249406.1	112
TNF-α	F：ACCTGGACTATGCCGAGTCT R：GAAGGGGATGAGGAGGGTCT	XM_012100437.4	127
IFN-γ	F：AAGTTCTTGAACGGCAGCTCTGAG R：TGAGGTTAGATTTTGGCGACAGGTC	NM_001009803.1	142
Occludin	F：GCCTGTGTTGCCTCCACTCTTG R：CCGTAGCCATAACCATAGCCATAGC	XM_015101255.3	131
ZO-1	F：TCCTGATCCTGAACCTGTGTCTGAG R：CTCCCGACTGGCACTCCTATCTC	XM_042235166.1	139
Claudin-1	F：TTCCACCGTAATGACCCTCCAGAG R：AAGGCACATCATTCCACCACAAGAG	XM_042230568.1	110
Claudin-4	F：ACCAGCAAGGAGGAATGGAAAG R：CAAGGGCAGGTAGGTGAGCAA	NM_001185017.2	297
CXCL8	F：GCTGGCTGTTGCTCTCTTGGC R：GGGGTGGAAAGGTGTGGAATGTG	NM_001009401.2	127
CXCL9	F：CAACACCAGCCAAGGGATGATCC R：CTTGAGCCCCATTCTTCACTGTAGC	XM_004009924.5	116
GAPDH	F：CCACGGCACAGTCAAGGCAGAGAAC R：CTCAGCACCAGCATCACCCCACTTG	NM_001190390.1	112

时间 Time/h	LPS浓度 LPS concentration/（µg/mL）						P值 P-value
时间 Time/h	0	1	5	10	50	100	P值 P-value
1	0.90±0.01^Ca	0.89±0.02^Aab	0.85±0.01^ABbc	0.88±0.01^Babc	0.85±0.01^Bc	0.86±0.01^Bbc	0.040
3	0.97±0.01^AB	0.95±0.02^A	0.90±0.02^A	0.94±0.02^A	0.91±0.03^A	0.92±0.02^A	0.250
6	0.94±0.02^ABa	0.84±0.01^Ab	0.81±0.01^Bb	0.81±0.01^Cb	0.81±0.02^Bb	0.82±0.01^Bb	<0.001
9	0.98±0.01^Aa	0.63±0.01^Bb	0.53±0.02^Cc	0.51±0.02^Dc	0.50±0.01^Cc	0.36±0.01^Cd	<0.001
18	0.93±0.01^BCa	0.38±0.05^Cb	0.36±0.03^Dbc	0.31±0.01^Ebc	0.31±0.04^Dbc	0.26±0.02^Dc	<0.001
P值 P-value	0.006	<0.001	<0.001	<0.001	<0.001	<0.001

时间 Time/h	芦丁浓度 Rutin concentration/（µg/mL）					P值 P-value
时间 Time/h	0	25	50	100	150	P值 P-value
0	0.95±0.01^BCab	0.95±0.03^a	0.94±0.01^Bab	0.90±0.02^b	0.93±0.01^ab	0.188
2	0.95±0.01^BC	0.95±0.01	0.96±0.01^B	0.95±0.01	0.96±0.01	0.938
4	1.05±0.02^ABab	1.06±0.01^a	1.05±0.01^ABab	1.05±0.02^ab	1.00±0.03^b	0.168
6	1.08±0.01^A	1.06±0.03	1.05±0.02^A	1.14±0.04	1.14±0.04	0.131
8	1.17±0.01^Ab	1.21±0.02^b	1.20±0.09^Ab	1.43±0.06^a	0.84±0.02^c	<0.001
10	0.75±0.04^D	0.75±0.05	0.74±0.02^C	0.74±0.03	0.74±0.04	0.998
P值 P-value	<0.001	<0.001	0.934	0.637	<0.001

项目 Items	组别 Groups				P值 P-value
项目 Items	CON	LPS	R	R+LPS	P值 P-value
白细胞介素-1β IL-1β	1.32±0.06^b	2.72±0.15^a	0.86±0.04^c	1.52±0.08^b	<0.001
白细胞介素-4 IL-4	1.07±0.03^b	0.63±0.03^d	1.45±0.08^a	0.83±0.05^c	<0.001
白细胞介素-6 IL-6	1.03±0.09^b	1.26±0.08^a	0.96±0.02^b	0.85±0.04^b	0.001
白细胞介素-10 IL-10	1.06±0.11^a	0.75±0.06^b	1.26±0.11^a	1.07±0.08^a	0.005
白细胞介素-12 IL-12	1.17±0.05^b	1.67±0.09^a	0.87±0.04^c	0.85±0.03^c	<0.001
肿瘤坏死因子-α TNF-α	1.14±0.02^b	1.90±0.02^a	0.97±0.08^c	1.26±0.06^b	<0.001
γ-干扰素 IFN-γ	1.03±0.01^b	2.03±0.20^a	0.82±0.06^b	1.01±0.02^b	<0.001
CXC趋化因子配体8 CXCL8	1.05±0.06^bc	1.45±0.04^a	0.96±0.04^c	1.15±0.08^b	<0.001
CXC趋化因子配体9 CXCL9	1.09±0.09^b	1.79±0.22^a	0.74±0.05^c	0.80±0.04^c	0.030

项目 Items	组别 Groups				P值 P-value
项目 Items	CON	LPS	R	R+LPS	P值 P-value
闭合蛋白-1 Claudin-1	1.01±0.05^b	0.80±0.04^b	1.93±0.14^a	1.07±0.15^b	<0.001
闭合蛋白-4 Claudin-4	1.05±0.03^b	0.64±0.06^c	1.85±0.04^a	1.01±0.05^b	<0.001
咬合蛋白 Occludin	1.01±0.04^b	0.70±0.07^b	1.48±0.18^a	0.98±0.06^b	<0.001
闭锁小带蛋白-1 ZO-1	1.12±0.09^b	0.50±0.08^c	1.89±0.40^a	1.05±0.04^b	<0.001

Effects of Rutin on Barrier Function and Inflammatory Injury of Rumen Epithelial Cells Induced by Lipopdysaccharide

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