[1]谢陆莉,李 鲲,邓 琳.岩黄连总碱下调miR-223表达调控人牙龈成纤维细胞增殖与凋亡的分子机制研究[J].陕西医学杂志,2023,52(11):1493-1497.[doi:DOI:10.3969/j.issn.1000-7377.2023.11.009]
　XIE Luli,LI Kun,DENG Lin.Molecular mechanism of corydalis saxicola bunting total alkaloids regulating roliferation and apoptosis of human gingival fibroblasts by down-regulating expression of miR-223[J].,2023,52(11):1493-1497.[doi:DOI:10.3969/j.issn.1000-7377.2023.11.009]
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岩黄连总碱下调miR-223表达调控人牙龈成纤维细胞增殖与凋亡的分子机制研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
52
期数:
2023年11期
页码:
1493-1497
栏目:
基础研究
出版日期:
2023-11-05

文章信息/Info

Title:
Molecular mechanism of corydalis saxicola bunting total alkaloids regulating roliferation and apoptosis of human gingival fibroblasts by down-regulating expression of miR-223
作者:
谢陆莉; 李 鲲; 邓 琳
(成都市龙泉驿区第一人民医院口腔科,四川 成都610100)
Author(s):
XIE Luli; LI Kun; DENG Lin
(Department of Stomatology,the First People's Hospital of Longquanyi District Chengdu,Chengdu 610100,China)
关键词:
岩黄连总碱;  微小RNA-223;  磷脂酰肌醇-3-激酶/蛋白激酶B信号通路;  人牙龈成纤维细胞;  牙龈卟啉单胞菌;  脂多糖
Keywords:
Corydalis saxicola bunting total alkaloids;  miR-223;  Phosphoinositide 3-kinase/Protein kinase B;  Human gingival fibroblasts;  Porphyromonas gingivalis;  Lipopolysacharide
分类号:
R 781.41
DOI:
DOI:10.3969/j.issn.1000-7377.2023.11.009
文献标志码:
A
摘要:
目的:探讨岩黄连总碱对牙龈卟啉单胞菌脂多糖(Pg-LPS)诱导的人牙龈成纤维细胞增殖与凋亡的影响及其可能作用机制。方法:体外培养人牙龈成纤维细胞,分为对照组(NC组)、Pg-LPS组、5 μg/ml岩黄连总碱+Pg-LPS组、10 μg/ml岩黄连总碱+Pg-LPS组、20 μg/ml岩黄连总碱+Pg-LPS组、miR-NC+Pg-LPS组、miR-223+Pg-LPS组、anti-miR-NC+Pg-LPS组、anti-miR-223+Pg-LPS组、20 μg/ml岩黄连总碱+Pg-LPS+miR-NC组、20 μg/ml岩黄连总碱+Pg-LPS+miR-223组。采用EDU、流式细胞术分别检测细胞增殖与凋亡; RT-qPCR法检测miR-223的表达量; ELISA法检测白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平; Western blot检测剪切的半胱天冬氨酸蛋白酶3(Cleaved-caspase-3)、pro-caspase-3、磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)蛋白表达。结果:岩黄连总碱(5、10、20 μg/ml)和下调miR-223表达可显著升高Pg-LPS诱导的人牙龈成纤维细胞增殖活力和pro-caspase-3蛋白表达水平,降低细胞凋亡率、IL-1β和TNF-α水平以及Cleaved-caspase-3、p-PI3K/PI3K、p-AKT/AKT蛋白表达水平(均P<0.05)。过表达miR-223可逆转岩黄连总碱对Pg-LPS诱导的人牙龈成纤维细胞增殖、凋亡及PI3K/AKT通路的作用。结论:岩黄连总碱可通过下调miR-223表达而抑制PI3K/AKT信号通路从而促进Pg-LPS诱导的人牙龈成纤维细胞增殖,并抑制细胞凋亡和炎性反应。
Abstract:
Objective:To explore the effect of corydalis saxicola bunting total alkaloids on the proliferation and apoptosis of human gingival fibroblasts induced by porphyromonas gingivalis lipopolysaccharide(Pg-LPS)and its possible mechanism.Methods:Human gingival fibroblasts were cultured in vitro and divided into control group(NC group),Pg-LPS group,5 μg/ml corydalis saxicola bunting total alkaloids+Pg-LPS group,10 μg/ml corydalis saxicola bunting total alkaloids+Pg-LPS group,20 μg/ml corydalis saxicola bunting total alkaloids+Pg-LPS group,miR-NC+Pg-LPS group,miR-223+Pg-LPS group,anti miR-NC+Pg-LPS group,anti miR-223+Pg-LPS group,20 μg/ml corydalis saxicola bunting total alkaloids+Pg-LPS+miR-NC group,20 μg/ml corydalis saxicola bunting total alkaloids+Pg-LPS+miR-223 group.Cell proliferation and apoptosis were detected by EDU and flow cytometry; the expression of miR-223 was detected by qRT-PCR; the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were determined by ELISA.Western blot was used to detect the expression of Cleaved caspase-3,pro-caspase-3,phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)proteins.Results:The corydalis saxicola bunting total alkaloids(5,10,20 μg/ml)and down-regulation of miR-223 expression significantly increased the proliferation activity and pro-caspase-3 protein level of Pg-LPS-induced human gingival fibroblasts,decreased apoptosis rate,IL-1β and TNF-α levels,and Cleaved caspase-3,p-PI3K/PI3K,p-AKT/AKT protein levels(all P<0.05).Overexpression of miR-223 reversed the effects of corydalis saxicola bunting total alkaloids on Pg-LPS-induced proliferation,apoptosis and PI3K/AKT pathway of human gingival fibroblasts.Conclusion:Corydalis saxicola bunting total alkaloids can inhibit the PI3K/AKT signaling pathway by downregulating the expression of miR-223,thereby promoting the proliferation of human gingival fibroblasts induced by Pg-LPS,and inhibiting cell apoptosis and inflammatory reactions.

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备注/Memo

备注/Memo:
基金项目:四川省卫生和计划生育委员会科研课题(16PJ091)
更新日期/Last Update: 2023-11-06