摘要
目的从PI3K/Akt/NF-κB信号通路探讨黄芩苷对脂多糖(LPS)诱导的人牙龈成纤维细胞(HGFs)损伤的影响及机制。方法运用LPS诱导建立人牙龈成纤维细胞损伤模型,设置正常对照组、模型组和模型+黄芩苷(1、10和20μg/mL)组,每组设3个复孔,经不同浓度的黄芩苷干预后,Western blotting检测p-Akt、Akt、NF-κB p65等蛋白的表达,采用qPCR法检测炎性因子TNF-α、IL-1β、IL-6等基因的表达。用PI3K抑制剂LY294002作用半小时后,再用黄芩苷干预,qPCR和Western blotting法检测Akt、p-Akt、NF-κB p65、TNF-α、IL-1β、IL-6等基因和蛋白的表达。结果与模型组比较,黄芩苷低、中、高剂量能有效降低TNF-α、IL-6、IL-1βmRNA的表达水平,促进p-Akt蛋白表达,抑制NF-κB p65核蛋白表达,并呈剂量依赖性(均P<0.05)。与模型组比较,模型+黄芩苷组p-Akt表达升高(P<0.05),NF-κB p65表达降低(P<0.01)。PI3K抑制剂LY294002作用后,模型+黄芩苷组p-Akt/Akt表达量(0.63±0.18)较模型组(0.56±0.14)升高(P<0.05),模型+黄芩苷组NF-κB p65、IL-6、IL-1β、TNF-α表达量较模型组降低(均P<0.01)。黄芩苷对LY294002作用后,p-Akt/Akt、NF-κB p65、IL-6、IL-1β、TNF-α表达量与模型组比较,差异无统计学意义。结论黄芩苷可抑制LPS诱导的人牙龈成纤维细胞损伤引起的炎症反应,其作用机制可能与促进Akt的磷酸化,抑制NF-κB p65核转录和炎性因子的释放有关。
Objective To investigate the effects of baicalin on the injury of human gingival fibroblasts(HGFs)induced by Lipopolysaccharide(LPS)through PI3 K/Akt/NF-κB signaling pathway and its mechanism.Methods LPS induced HGFs injury models were established and divided into normal control group,model group and model+baicalin(1,10 and 20μg/mL)groups.Each group was set with 3 repetitions.After treatment with different concentrations of baicalin,Western blotting was used to detect the expression of p-Akt,Akt and NF-κB p65,and the expressions of inflammatory factors TNF-α,IL-1βand IL-6 were detected by qPCR.After treated with PI3 K inhibitor LY294002 for half an hour,baicalin was used to intervene,and the genes and proteins expressions of Akt,p-Akt,NF-κB p65,TNF-α,IL-1βand IL-6 were detected by qPCR and Western blotting.Results Compared with the model group,low,medium and high dose baicalin could effectively reduce the mRNA expression levels of TNF-α,IL-6 and IL-1β,promote the protein expression of P-Akt and inhibit the nuclear protein expression of NF-κB p65 in a dose-dependent manner(all P<0.05).Compared with the model group,the expression of p-Akt in the model+baicalin group was increased(P<0.05),and the expression of NF-κB p65 was decreased(P<0.01).After the treatment of PI3 K inhibitor LY294002,the expression level of p-Akt/Akt in the model+baicalin group was increased(0.63±0.18)compared with that in the model group(0.56±0.14,P<0.05),and the expressions of NF-κB p65,IL-6,IL-1βand TNF-αin the model+baicalin group were decreased compared with those in the model group(all P<0.01).After baicalin treated LY294002,the expression levels of p-Akt/Akt,NF-κB p65,IL-6,IL-1βand TNF-αwere not significantly different from those of the model group.Conclusion Baicalin can inhibit the inflammatory response induced by LPS-induced human gingival fibroblast injury,and its mechanism may be related to promote the phosphorylation of Akt,inhibit NF-κB p65 nuclear transcription and the release of inflammatory factors.
作者
高娟
伍燕
郑之峻
王青云
刘曙
党妮
GAO Juan;WU Yan;ZHENG Zhi-jun;WANG Qing-yun;LIU Shu;DANG Ni(Department of Orthodontics,Guiyang Stomatological Hospital,Guiyang,Guizhou 550002,China;不详)
出处
《中华全科医学》
2022年第12期2002-2005,共4页
Chinese Journal of General Practice
基金
贵州省卫生健康委科学技术基金项目(gzwjkj2020-1-162)。
关键词
黄芩苷
脂多糖
人牙龈成纤维细胞
炎症反应
Baicalin
Lipopolysaccharide
Human gingival fibroblasts
Inflammatory response
