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

doi:10.11958/20221590

Abstract

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

CLC Number:

R576

Figures/Tables 7

References 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^＊＊