Lv Jia-Shu, Jiang Xue-Wei, Zhang Yan, Zhen Lei
Department of Periodontology, Shanghai Stomatological Disease Center. Shanghai 200001, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2017 Feb;26(1):32-36.
Through a study of the molecular mechanism of the effect of resveratrol(RSV) on expression of TLR4 and inflammatory factors in gingival epithelial cells under high glucose environment, the therapeutic effect and molecular mechanism of resveratrol on periodontitis in patients with diabetes mellitus was investigated.
Gingival epithelial cells were cultured in vitro; according to the way of action, the cultured cells were divided into control group, high glucose group(HG) and HG+RSV group. The mRNA expression of TLR4 was detected by PCR; The third generation of gingival epithelial cells were pre-treated with or without RSV for 24 h under high glucose conditions, and subsequently treated with LPS at 100 ng/mL for 2 h. ELISA was used to detect the secretion of IL-1 beta, IL-6, IL-8 and TNF- alpha; the activation of TLR4 downstream signaling molecules NF-κB p65, p38 MAPK, and STAT3 was determined by Western blot. SPSS17.0 software package was used for statistical analysis.
RSV could reverse the increase of TLR4 level in gingival epithelial cells in high glucose medium.LPS markedly increased the expression and secretion of IL-1β, IL-6, IL-8, and TNF-α in GECs cultured in high glucose medium, which was partly blocked in the presence of RSV. Furthermore, Western blot results showed that RSV significantly suppressed the phosphorylation of TLR4 downstream factors NF-κB p65, p38MAPK, and STAT3.
RSV reduces inflammatory cytokine secretion in gingival epithelial cells, through negative regulation of TLR4 signaling pathway.
通过研究白藜芦醇(RSV)对高糖环境下牙龈上皮细胞TLR4表达及炎症因子的影响的分子机制,探讨白藜芦醇对糖尿病患者牙周炎的治疗作用及分子机制。
体外培养牙龈上皮细胞;根据作用方式,将培养的细胞分为对照组、高糖组(HG)和HG+RSV组。采用PCR检测TLR4的mRNA表达;第三代牙龈上皮细胞在高糖条件下用或不用RSV预处理24 h,随后用100 ng/mL LPS处理2 h。采用ELISA检测IL-1β、IL-6、IL-8和TNF-α的分泌;通过Western blot检测TLR4下游信号分子NF-κB p65、p38 MAPK和STAT3的激活情况。使用SPSS17.0软件包进行统计分析。
RSV可逆转高糖培养基中牙龈上皮细胞TLR4水平的升高。LPS显著增加高糖培养基中培养的牙龈上皮细胞中IL-1β、IL-6、IL-8和TNF-α的表达和分泌,在RSV存在的情况下部分受到抑制。此外,Western blot结果显示,RSV显著抑制TLR4下游因子NF-κB p65、p38MAPK和STAT3的磷酸化。
RSV通过负调控TLR4信号通路减少牙龈上皮细胞中炎性细胞因子的分泌。