【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。更多还原

【Abstract】 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 20 μg/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-1β and 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-1β and 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-1β and TNF-α in 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-1β and TNF-α were 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.更多还原