miR-143-3p靶向调控TLR2/ NF-κB/NLRP3通路对溃疡性结肠炎细胞焦亡的影响

doi:10.3969/j.issn.1006-5725.2024.15.003

Abstract

Abstract:

Objective To study the effect of miR-143-3p on pyroptosis of HCT-116 cells induced by LPS + ATP. Methods The targeting relationship between miR-143-3 p and TLR2 gene was detected by dual luciferase. The pyroptosis model was established after transfection of miR-143-3p mimic. Cell apoptosis was detected by flow cytometry， and the activity of Caspase-1 and LDH was detected by biochemical method. The levels of IL-1β and IL-18 were detected by ELISA. The mRNA levels of NF-κB， TLR2， NLRP3 and GSDMD were detected by q-PCR. The expression of NLRP3 and ASC was detected by immunofluorescence. The protein expressions of TLR2， GSDMD， p-NF-κB p65 and cleave Caspase-1 were detected by Western blot. Results Dual luciferase assay showed that miR-143-3p targeted TLR2 expression. The expressions of NF-κB， TLR2， NLRP3， GSDMD mRNA and TLR2， GSDMD， p-NF-κB p65， cleave Caspase-1， ASC， NLRP3 protein in miR-143-3p mimic group and si TLR2 group were lower than those in model group. The activity of Caspase-1 and the content of LDH， IL-1β and IL-18 decreased. Conclusion miR-143-3p regulates pyroptosis of ulcerative colitis cells by targeting TLR2 gene and regulating NF-κB/NLRP3/Caspase-1 signaling pathway.

Key words: ulcerative colitis, cell pyroptosis, miR-143-3p, TLR2, TLR2/ NF-κB/NLRP3

CLC Number:

R259

Xiuli SHI,Jiaqi CHEN,Fan ZHU,Juan ZENG,Na. WU. The effect of miR⁃143⁃3p on pyroptosis of ulcerative colitis cells by regulating TLR2/NF⁃κB/NLRP3[J]. The Journal of Practical Medicine, 2024, 40(15): 2056-2062.

Figures/Tables 8

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References 31

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引物名称	引物序列（5′-3′）	长度（bp）
U6	上游：CGCTTCGGCAGCACATATAC
U6	下游：AAATATGGAACGCTTCACGA
miR-143-3p	上游：GTCGTATCCAGTGCAGGGTCCGAGGTATTCG CACTGGATACGACGAGCTACA
miR-143-3p	下游：TGCGCTGAGATGAAGCACTGT
GAPDH	上游：GGAAGCTTGTCATCAATGGAAATC	168
GAPDH	下游：TGATGACCCTTTTGGCTCCC	168
GSDMD	上游：TGGTTATTGACTCTGACTTGGACG	164
GSDMD	下游：ATCTGTCAGGAAGTTGTGGAGGC	164
TLR2	上游：TGTGAAGAGTGAGTGGTGCA	100
TLR2	下游：TACCCAAAATCCTTCCCGCT	100
NLRP3	上游：TGGTCTGCTGGATCGTGTGC	84
NLRP3	下游：CGGTGGTGGTCTTGGATGTCTG	84
NF-κB	上游：TCCCATCTTTGACAATCGTGC	155
NF-κB	下游：AGCCTGGTCCCGTGAAATAC	155

分组	海肾/萤火虫
空白组	0.92 ± 0.07
mimics NC+野生型TLR2 3'UTR组	1.43 ± 0.01^?
miR-143-3p mimics+野生型TLR2 3'UTR组	0.59 ± 0.02^?#
mimics NC+突变型TLR2 MUT 3'UTR组	1.42 ± 0.02^?
miR-143-3p mimics+突变型TLR2 MUT 3'UTR组	1.43 ± 0.36^?

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The effect of miR⁃143⁃3p on pyroptosis of ulcerative colitis cells by regulating TLR2/NF⁃κB/NLRP3

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