目的 观察银连含漱液对脂多糖（LPS）诱导人牙龈成纤维细胞（HGFs）炎症反应、氧化应激及细胞凋亡的 影响。方法 收集临床患者的健康牙龈组织，采用组织块法培养HGFs并采用免疫荧光法进行细胞鉴定。采用 CCK-8法检测不同体积分数银连含漱液（2.5%、5%、10%、20%、40%）、不同浓度绿原酸（0.01、0.1、0.5、1、 10 mg·mL-1）及LPS（1、10、20、35、50 μg·mL-1）分别与HGFs共培养24、48、72 h后的增殖变化情况。依据CCK-8 检测结果，将 HGFs 分为 4 组：DMEM 对照组、1 μg·mL-1 LPS 组、1 μg·mL-1 LPS +20% 银连含漱液组、 1 μg·mL-1 LPS +1 mg·mL-1绿原酸组。干预 24 h后，采用流式细胞术检测 HGFs的凋亡水平；WST-1法检测细 胞超氧化物歧化酶（SOD）活性；ELISA 法检测细胞上清液中炎性因子肿瘤坏死因子 α（TNF-α）、白细胞介 素 6（IL-6）的含量。结果 选择对HGFs无明显毒性作用的1 mg·mL-1绿原酸、20%银连含漱液和1 μg·mL-1 LPS 作用24 h。与对照组比较，LPS组的HGFs凋亡率显著升高（P＜0.001），SOD活力显著降低（P＜0.001），上清液 中的IL-6、TNF-α水平显著升高（P＜0.001）。与LPS组比较，LPS+20%银连含漱液组及LPS+1 mg·mL-1绿原酸 组的HGFs凋亡率显著降低（P＜0.001），SOD活力显著升高（P＜0.01，P＜0.001），上清液中的IL-6、TNF-α水平 显著降低（P＜0.001）。与 LPS+20% 银连含漱液组比较，LPS+1 mg·mL-1绿原酸组的 HGFs 凋亡率显著降低（P＜ 0.001），SOD活力显著升高（P＜0.01），上清液中的IL-6、TNF-α水平显著降低（P＜0.05，P＜0.01）。结论 银 连含漱液可抑制LPS诱导的HGFs细胞凋亡，提高其抗氧化能力，降低炎症反应，该作用可能与活性成分绿原 酸密切相关。

Objective To study the effects of Yinlian Gargle on inflammation，oxidative stress response，and apoptosis of human gingival fibroblasts（HGFs）stimulated by lipopolysaccharide（LPS）. Methods Healthy gingival samples were collected from the donors. HGFs were cultured by tissue-explant technique and identified by immunofluorescence assay. CCK-8 method was used to observe the effects of different volume fractions（2.5%，5%，10%，20%，40%）of Yinlian Gargle，different concentrations of chlorogenic acid（0.01，0.1，0.5，1，10 mg·mL-1）and different concentrations of LPS（1， 10， 20， 35， 50 μg·mL-1）on cell proliferation of HGFs at different time duration （24， 48， 72 h）. According to CCK-8 results，HGFs were divided into four groups：DMEM control group，1 μg·mL-1 LPS group， 1 μg·mL-1 LPS +20% Yinlian Gargle group， and 1 μg·mL-1 LPS +1 mg·mL-1 chlorogenic acid group. After 24 h of intervention， flow cytometry， WST-1 assay and enzyme-linked immunosorbent assay（ELISA）were used to test apoptosis of HGFs，superoxide dismutase（SOD）activity，TNF-α and IL-6 levels. Results No significant toxic effect on HGFs was observed by 24 h treatment of 20% of Yinlian Gargle，1 mg·mL-1 of chlorogenic acid and 1 μg·mL-1 of LPS，which were chosen for subsequent experiments. Compared to the control group，the apoptosis rate of HGFs in LPS group significantly increased（P＜0.001），SOD activity significantly decreased（P＜0.001），while the levels of IL-6 and TNF- α in the supernates significantly increased（P＜0.001）. Compared to LPS group， the apoptosis rate of 1 μg·mL-1 LPS +20% Yinlian Gargle group， and 1 μg·mL-1 LPS +1 mg·mL-1chlorogenic acid group significantly decreased（P＜0.001），SOD activities significantly increased（P＜0.01，P＜0.001），while the levels of IL-6 and TNF-α in the supernates significantly decreased（P＜0.001）. Compared to 1 μg·mL-1 LPS +20% Yinlian Gargle group， the apoptosis rate of HGFs in 1 μg·mL-1 LPS +1 mg·mL-1chlorogenic acid group significantly decreased（P＜0.001）， SOD activity significantly increased（P＜0.01），while the levels of IL-6 and TNF- α in the supernates significantly decreased（P＜0.05，P＜0.01）. Conclusion Yinlian Gargle could inhibit LPS-induced apoptosis of HGFs，enhance antioxidant capacity，reduce inflammatory response，which may be closely related to chlorogenic acid.

R285.5

广东省中医药局科研项目（20231168）