Ropivacaine alleviates LPS-induced apoptosis of ulcerative colitis cell line NCM-460
10.16352/j.issn.1001-6325.2024.10.1368
- VernacularTitle:罗哌卡因减轻LPS诱导的人结肠上皮细胞系NCM-460凋亡
- Author:
Lingqin ZHOU
1
;
Weijuan WANG
;
Lingling REN
;
Junlai ZHU
;
Guanglan CHEN
Author Information
1. 丽水市第二人民医院 麻醉科,浙江 丽水 323000
- Keywords:
human colon epithelial cell line;
lipopolysaccharide;
ropivacaine;
proliferation;
apoptosis
- From:
Basic & Clinical Medicine
2024;44(10):1368-1375
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impact of ropivacaine on apoptosis of lippolysaccharide(LPS)-induced ulcerative colitis cell line NCM-460 and on activity of nucleotide oligomerization domain(NOD)-like receptor protein-3(NLRP3)inflammatome.Methods Human colon epithelial cell line NCM-460 was cultured in vitro and divided into control group(no intervention),model group(10 μg/mL LPS treatment),low/medium/high concentration ropivacaine group(10 μg/mL LPS and 0.5,1,1.5 mmol/L ropivacaine co-treatment,respectiv-oly).Cell viability was determined by cell counting kit 8(CCK-8)and the appropriate concentration was selected.The cells were then divided into control group,model group,ropivacaine group(10 μg/mL LPS and 1.5 mmol/L ropivacaine treatment)and ropivacaine+inhibitor group(10 μg/mL LPS,1.5 mmol/L ropiva-caine and 1 μmol/L NF-κB pathway inhibitor BAY 11-7082 treatment),inhibitor group(10 μg/mL LPS+1 μmol/L NF-κB pathway inhibitor BAY 11-7082 treatment)and ropivacaine+activator group(10 μg/mL LPS,1.5 mmol/L ropivacaine and 1 μmol/L NF-κB pathway activator Prostratin),all groups were treated for 24 h.The level of IL-6,IL-8 and TNF-α were detected by enzyme-linked immunosorbent assay(ELISA).The proliferation rate was detected by EdU incorporation.Hoechst 33258 staining microscopy was used to detect the apoptosis rate.Level of cyclinD1,caspase-3,NLRP3 and NF-κB pathway-related proteins were detected by Western blot.Results Compared with the control group,the cell viability of the model group was significantly decreased and the cell viability of high-concentration experimental group was increased after adding ropivacaine(P＜0.05).So,1.5 mmol/L ropivacaine was selected for the follow-up experiment.Compared with the control group,the concentration of inflammatory cytokines IL-6,IL-8,TNF-α,apoptosis rate and the protein expression of caspase-3,NLRP3 and phosphorylated p-NF-κB in model group were all significantly increased(P＜0.05),while the proliferation rate and cycilnD1 protein expression were decreased(P＜0.05).Com-pared with model group,the concentrations of IL-6,IL-8,TNF-α,apoptosis rate and the expression of caspase-3,NLRP3 and p-NF-κB protein in ropivacaine group and inhibitor group were significantly decreased(P＜0.05),while the proliferation and cycilnD1 protein expression were increased(P＜0.05).Compared with ropivacaine group,the trend of the above indexes in ropivacaine+inhibitor group was more significant(P＜0.05),and the trend of these indexes in ropivacaine+agonist group was significantly reversed(P＜0.05).Conclusions Ropivacaine can inhibit the activation of NLRP3 inflammasome and block the signal transduction of NF-κB pathway,further inhibit LPS-induced apoptosis of NCM-460 cells and promote proliferation.
