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目的 :研究白藜芦醇对高糖环境下牙龈上皮细胞TLR4表达及炎症因子分泌的影响,探讨白藜芦醇对伴糖尿病牙周炎患者的治疗作用及相关分子机制。方法:体外培养牙龈上皮细胞,按照作用方式不同分为正常对照组、高糖组和高糖+白藜芦醇组。荧光定量PCR检测TLR4表达情况;取第3代牙龈上皮细胞,高糖环境下采用或不采用白藜芦醇处理24 h,随后用100 ng/m L LPS处理2 h,ELISA检测IL-1β、IL-6、IL-8和TNF-α分泌;Western印迹法检测TLR4信号通路下游分子NF-κB p65、p38MAPK和STAT3磷酸化。采用SPSS 17.0软件包对数据进行统计学分析。结果:白藜芦醇可以逆转高糖环境下牙龈上皮细胞TLR4水平的升高,同时抑制高糖环境下LPS诱导的IL-1β、IL-6、IL-8和TNF-α表达。Western印迹结果显示,白藜芦醇也可抑制TLR4信号通路下游分子NF-κB p65、p38MAPK和STAT3磷酸化。结论 :白藜芦醇通过负向调控TLR4信号通路减轻高糖环境下牙龈上皮细胞炎症因子的分泌。
Objective: To investigate the effects of resveratrol on the expression of TLR4 and the secretion of inflammatory cytokines in gingival epithelial cells under high glucose conditions and to explore the therapeutic effect and related molecular mechanisms of resveratrol on patients with diabetes mellitus. Methods: Gingival epithelial cells were cultured in vitro and divided into normal control group, high glucose group and high glucose + resveratrol group according to the mode of action. Fluorescent quantitative PCR was used to detect the expression of TLR4. The 3rd generation of gingival epithelial cells were treated with or without resveratrol in high glucose for 24 h, then treated with 100 ng / mL LPS for 2 h, and the levels of IL-1β, IL-6, IL-8 and TNF-α secretion; phosphorylation of NF-κB p65, p38MAPK and STAT3 in TLR4 signaling pathway were detected by Western blot. Data were statistically analyzed using SPSS 17.0 software package. RESULT: Resveratrol can reverse the increase of TLR4 level in gingival epithelial cells under high glucose and inhibit the expression of IL-1β, IL-6, IL-8 and TNF-α induced by LPS in high glucose. Western blotting results showed that resveratrol could also inhibit the phosphorylation of NF-κB p65, p38MAPK and STAT3 downstream of TLR4 signaling pathway. CONCLUSION: Resveratrol can reduce the secretion of inflammatory cytokines in gingival epithelial cells under high glucose condition by negatively regulating TLR4 signaling pathway.