尿石素A通过TLR4/NF-κB信号通路对重症急性胰腺炎的保护作用及机制

doi:10.11958/20221590

天津医药 ›› 2023, Vol. 51 ›› Issue (6): 613-617.doi: 10.11958/20221590

尿石素A通过TLR4/NF-κB信号通路对重症急性胰腺炎的保护作用及机制

曾家月¹(), 马昕玥¹, 马凤雨¹, 陈霞¹^,²^,^△

1 西南医科大学附属医院消化内科（邮编646000）
2 成都医学院第一附属医院消化内科

收稿日期:2022-09-28 修回日期:2022-12-16 出版日期:2023-06-15 发布日期:2023-06-20
通讯作者: ^△E-mail：970217858@qq.com
作者简介:曾家月（1995），女，硕士在读，主要从事急性胰腺炎的基础和临床方面研究。E-mail：20200299120017@stu.swmu.edu.cn
基金资助:
国家自然科学基金资助项目(81600420);泸州市人民政府-西南医科大学联合项目(2019LZXNYDJ48)

The protective effect of urolithin A on severe acute pancreatitis through TLR4 / NF-κB signaling pathway

ZENG Jiayue¹(), MA Xinyue¹, MA Fengyu¹, CHEN Xia¹^,²^,^△

1 Department of Gastroenterology, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, China
2 Department of Gastroenterology, the First Affiliated Hospital of Chengdu Medical College

Received:2022-09-28 Revised:2022-12-16 Published:2023-06-15 Online:2023-06-20
Contact: ^△E-mail：970217858@qq.com

曾家月, 马昕玥, 马凤雨, 陈霞. 尿石素A通过TLR4/NF-κB信号通路对重症急性胰腺炎的保护作用及机制[J]. 天津医药, 2023, 51(6): 613-617.

ZENG Jiayue, MA Xinyue, MA Fengyu, CHEN Xia. The protective effect of urolithin A on severe acute pancreatitis through TLR4 / NF-κB signaling pathway[J]. Tianjin Medical Journal, 2023, 51(6): 613-617.

摘要/Abstract

摘要：

目的探讨尿石素A（UroA）对重症急性胰腺炎（SAP）大鼠胰腺损伤的保护作用及机制。方法 36只健康雄性SD大鼠随机分为假手术组（Sham组）、SAP组及UroA组，每组12只。采用逆行胆胰管注射5%牛磺胆酸钠溶液的方法构建SAP大鼠模型。UroA组于SAP造模后灌胃UroA溶液（10 mg/kg），每6 h灌胃给药1次，共4次。建模24 h后，统计各组大鼠腹水量，检测血清脂肪酶（LPS）水平，酶联免疫吸附试验检测血清白细胞介素（IL）-6、肿瘤坏死因子（TNF）-α水平，HE染色观察胰腺组织病理损伤情况；实时荧光定量PCR分析胰腺组织IL-6、TNF-α、Toll样受体4（TLR4）、MyD88、核转录因子（NF）-κB mRNA表达水平；蛋白质免疫印迹法检测胰腺组织TLR4、MyD88、NF-κB p65蛋白表达水平。结果与Sham组相比，SAP组大鼠血清LPS、IL-6、TNF-α水平升高，腹水量及胰腺病理学评分增加（P<0.05），胰腺IL-6、TNF-α、TLR4、MyD88、NF-κB的mRNA水平升高（P<0.05），TLR4、MyD88、NF-κB p65的蛋白水平升高（P<0.05）；与SAP组比较，UroA组大鼠胰腺病理损伤改善，血清LPS、IL-6、TNF-α水平降低，TLR4、MyD88、NF-κB蛋白及mRNA水平降低（P<0.05）。结论 UroA应用可改善SAP大鼠胰腺病理损伤，其机制可能与抑制TLR4/NF-κB信号通路活化，从而抑制炎症反应有关。

关键词: 胰腺炎, Toll样受体4, NF-κB, 尿石素A, 重症急性胰腺炎

Abstract:

Objective To investigate the effect and its protective mechanism of urolithin A (UroA) on pancreatic injury in rats with severe acute pancreatitis (SAP). Methods Thirty-six healthy male SD rats were randomly divided into the sham group, the SAP group and the UroA group, with 12 rats in each group. The SAP model was established by retrograde perfusion of 5% sodium taurocholate into biliopancreatic duct. The UroA group was given intervention 10 mg/kg by UroA once 6 h for 4 times immediately after operation. Twenty-four hours after modeling, the ascites volume was counted. Meanwhile, the serum lipase (LPS) level was detected. The serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay, and the pathological injury of pancreatic tissue was detected by HE staining. The expression levels of IL-6, TNF-α, TLR4, MyD88 and NF-κB mRNA in pancreatic tissue were analyzed by RT-qPCR. The protein expressions of TLR4, MyD88 and NF-κB p65 in pancreatic tissue were detected by Western blot assay. Results Compared with the sham group, the serum levels of LPS, IL-6 and TNF-α increased in the SAP group (P<0.05), ascites volume and pancreatic pathological score increased (P<0.05) and the mRNA levels of IL-6, TNF-α, TLR4, MyD88, NF-κB and the protein expression of TLR4, MyD88, NF-κB p65 in pancreatic tissue were also up-regulated (P<0.05). Compared with the SAP group, the pancreatic pathological injury significantly improved in the UroA group. The serum levels of LPS, IL-6 and TNF-α were significant decreased. In addition, TLR4, MyD88, NF-κB protein and mRNA levels were decreased (P<0.05). Conclusion UroA can alleviate the pathological injury of pancreas in rats with SAP, and its mechanism may be related to inhibiting the activation of TLR4/NF-κB signaling pathway, and thereby inhibiting inflammatory response.

Key words: pancreatitis, Toll-like receptor 4, NF-kappa B, urolithin A, severe acute pancreatitis

中图分类号:

R576

图/表 7

参考文献 17

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基因名称	引物序列（5′→3′）	产物大小（bp）
TLR4	上游：TATCGGTGGTCAGTGTGCTT	104
TLR4	下游：CTCGTTTCTCACCCAGTCCT	104
MyD88	上游：TTCGACGCCTTCATCTGCTA	116
MyD88	下游：CATGCGACGACACCTTTTCT	116
NF-κB	上游：ACGCAAAAGGACCTACGAGA	97
NF-κB	下游：ATGGTGCTGAGGGATGTTGA	97
IL-6	上游：GGAGTTCCGTTTCTACCTGG	103
IL-6	下游：GCCGAGTAGACCTCATAGTG	103
TNF-α	上游：GCGTGTTCATCCGTTCTCTACC	167
TNF-α	下游：TACTTCAGCGTCTCGTGTGTTTCT	167
GAPDH	上游：CCATTCTTCCACCTTTGATGCT	181
GAPDH	下游：TGTTGCTGTAGCCATATTCATTGT	181

基因名称	引物序列（5′→3′）	产物大小（bp）
TLR4	上游：TATCGGTGGTCAGTGTGCTT	104
TLR4	下游：CTCGTTTCTCACCCAGTCCT	104
MyD88	上游：TTCGACGCCTTCATCTGCTA	116
MyD88	下游：CATGCGACGACACCTTTTCT	116
NF-κB	上游：ACGCAAAAGGACCTACGAGA	97
NF-κB	下游：ATGGTGCTGAGGGATGTTGA	97
IL-6	上游：GGAGTTCCGTTTCTACCTGG	103
IL-6	下游：GCCGAGTAGACCTCATAGTG	103
TNF-α	上游：GCGTGTTCATCCGTTCTCTACC	167
TNF-α	下游：TACTTCAGCGTCTCGTGTGTTTCT	167
GAPDH	上游：CCATTCTTCCACCTTTGATGCT	181
GAPDH	下游：TGTTGCTGTAGCCATATTCATTGT	181

组别	胰腺病理学评分（分）	血清LPS（U/L）
Sham组	3.10±0.96	55.19±9.94
SAP组	13.20±1.15^a	139.70±5.36^a
UroA组	9.90±0.65^ab	90.82±7.19^ab
F	148.729^＊＊	181.114^＊＊

组别	胰腺病理学评分（分）	血清LPS（U/L）
Sham组	3.10±0.96	55.19±9.94
SAP组	13.20±1.15^a	139.70±5.36^a
UroA组	9.90±0.65^ab	90.82±7.19^ab
F	148.729^＊＊	181.114^＊＊

组别	血清（n=5，ng/L）		胰腺组织（n=3）
组别	IL-6	TNF-α	IL-6	TNF-α
Sham组	28.49±5.50	83.98±4.14	1.24±0.46	1.00±0.06
SAP组	66.15±4.09^a	172.80±24.53^a	48.72±4.99^a	5.34±0.38^a
UroA组	39.55±3.61^ab	102.00±10.11^b	24.48±8.14^ab	1.78±0.06^ab
F	93.652^＊＊	45.869^＊＊	55.545^＊＊	319.328^＊＊

尿石素A通过TLR4/NF-κB信号通路对重症急性胰腺炎的保护作用及机制

The protective effect of urolithin A on severe acute pancreatitis through TLR4 / NF-κB signaling pathway

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组别	TLR4	MyD88	NF-κB
Sham组	1.01±0.02	1.01±0.20	0.99±0.02
SAP组	2.43±0.17^a	5.24±0.36^a	1.94±0.09^a
UroA组	1.12±0.17^b	3.78±0.12^ab	1.56±0.12^ab
F	94.740^＊＊	225.088^＊＊	86.994^＊＊

组别	TLR4	MyD88	NF-κB p65
Sham组	0.25±0.10	0.36±0.10	0.27±0.04
SAP组	0.95±0.16^a	0.91±0.06^a	0.98±0.07^a
UroA组	0.59±0.15^ab	0.49±0.12^b	0.42±0.09^ab
F	18.592^＊＊	27.074^＊＊	85.135^＊＊

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