摘要
目的探讨没食子水提取物对脂多糖诱导的人牙龈上皮角质形成细胞(HGEK)的抗炎作用,以及对细胞组织伤口愈合作用的影响。方法选取中国人民解放军空军军医大学第九八六医院口腔科2020年6月至12月收治的10例牙冠延长术患者的健康牙龈,体外培养HGEK细胞,使用第3~5代细胞。分为阴性对照组(A组,PBS),脂多糖组(B组,质量浓度为1μg/mL),低、中、高没食子水提取物组(分别为C_(1)组、C_(2)组、C_(3)组,没食子酸质量浓度分别为1,5,10 mg/mL),氯己定组(D组,蒸馏水调整20%葡萄糖酸氯己定质量分数至0.2%)。采用四甲基偶氮唑盐比色(MTT)法测定细胞活力;采用酶联免疫吸附(ELISA)法测定白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)基因和蛋白表达的水平;采用划痕愈合实验观察没食子水提取物对细胞迁移的影响;采用蛋白免疫印迹法测定核因子E2相关因子(Nrf2)和血红素加氧酶1(HO-1)基因和蛋白表达水平。两组间定量资料采用Bonferroni检验,多组间定量资料比较采用方差分析(ANOVA)。结果与A组比较,C_(2)组、C_(3)组细胞活力显著增强(P<0.05)。与B组比较,C_(2)组、C_(3)组IL-1β,IL-6,TNF-α基因和蛋白表达水平均显著降低(P<0.05),C_(3)组细胞迁移率显著增加(P<0.05);与A组比较,C_(3)组Nrf2和HO-1基因和蛋白的表达水平显著增强(P<0.05)。结论没食子水提取物可下调炎性细胞因子水平,减轻脂多糖诱导的HGEK的炎性反应,具有作为口服口腔抗炎药的可能性。
Objective To investigate the anti-inflammatory effect of gallic water extract on lipopolysaccharide(LPS)-induced human gingival epithelial keratinocytes(HGEK),and its effects on the wound healing of cells and tissue.Methods The healthy gums of 10 patients underwent crown lengthening surgery who were admitted to the Department of Stomatology of the No.986 Air Force Hospital of Air Force Medical University from June to December 2020 were selected.The HGEK cells were cultured in vitro and the third to fifth generation cells were used.They were divided into the negative control group(group A,PBS),LPS group(group B,with a mass concentration of 1μg/mL),low-,medium-and high-gallic water extract groups(groups C_(1),C_(2) and C_(3),with the mass concentration of gallic acid of 1,5 and 10 mg/mL,respectively)and chlorhexidine group(group D,distilled water adjusted the mass fraction of 20%chlorhexidine gluconate to 0.2%).Cell viability was determined by the methylthiazoletrazolium(MTT)colorimetry assay.The expression levels of interleukin 1β(IL-1β),interleukin 6(IL-6),tumor necrosis factor-α(TNF-α)genes and proteins were determined by the enzyme-linked immunosorbent assay(ELISA).The effect of gallic water extract on cell migration was observed by the scratch wound healing assay.The expression levels of nuclear factor E2 related factor(Nrf2)and heme oxygenase-1(HO-1)genes and proteins were determined by the Western blot.Quantitative data between two groups was analyzed by the Bonferroni test,and quantitative data among multiple groups was compared by the analysis of variance(ANOVA).Results Compared with those in group A,the cell viability in group C_(2) and group C_(3) enhanced significantly(P<0.05).Compared with those in group B,the expression levels of IL-1β,IL-6 and TNF-αgenes and proteins in group C_(2) and group C_(3) decreased significantly(P<0.05),and the cell migration rate in group C_(3) increased significantly(P<0.05).Compared with those in group A,the expression levels of Nrf2 and HO-1 genes and proteins of group C_(3) enhanced significantly(P<0.05).Conclusion The gallic water extract can down-regulate the level of inflammatory cytokines,alleviate the inflammatory response of HGEK induced by LPS and has the possibility of being used as an oral anti-inflammatory drug.
作者
徐娜
李恺
高鹏
张若冰
艾林
XU Na;LI Kai;GAO Peng;ZHANG Ruobing;AI Lin(No.986 Air Force Hospital of Air Force Medical University,Xi'an,Shaanxi,China 710054;Shaanxi Medical Products Administration,Xi'an,Shaanxi,China 710065)
出处
《中国药业》
CAS
2022年第9期60-64,共5页
China Pharmaceuticals
基金
中国人民解放军“十三五”军队后勤科研计划项目[CKJ20J027]。
关键词
没食子水提取物
人牙龈上皮角质形成细胞
脂多糖
炎性细胞因子
抗炎作用
作用机制
划痕伤口愈合实验
gallic water extract
human gingival epithelial keratinocytes
lipopolysaccharide
inflammatory cytokine
anti-inflammatory effect
mechanism
scratch wound healing assay
