[摘 要] 目的：探究连翘苷（PHN）调节高迁移率族蛋白 B1（HMGB1）/晚期糖基化终产物受体（RAGE）信号通路对胶质瘤 U251细胞增殖、侵袭及上皮间质转化（EMT）的影响。方法：将人胶质瘤U251细胞分为PHN-0组（0 μmol/L PHN处理）、PHN低、 中和高剂量组（PHN-50、PHN-100、PHN-200组，分别用50、100和200 μmol/L PHN处理）、PHN + pcDNA-NC组（转染pcDNA-NC 质粒后200 μmol/L PHN处理）和PHN + HMGB1组（转染过表达HMGB1质粒后200 μmol/L PHN处理）。CCK-8法和克隆形成实 验检测各组细胞的增殖能力，流式细胞术检测各组细胞的凋亡水平，Transwell实验检测各组细胞的迁移和侵袭能力，ELISA检测 各组细胞分泌IL-8水平，免疫荧光法检测各组细胞中神经钙黏素（N-cadherin）和上皮钙黏素（E-cadherin）阳性率，WB法检测各组 细胞中Toll样受体4（TLR4）、核因子-κB（NF-κB）、HMGB1、RAGE、N-cadherin、E-cadherin、细胞周期蛋白D1（cyclin D1）、细胞周 期蛋白依赖性激酶2（CDK2）、B淋巴细胞瘤-（2 Bcl-2）、Bcl-2相关X蛋白（BAX）蛋白的表达水平。结果：与PHN-0组相比，PHN-50、 PHN-100、PHN-200 组 U251 细胞增殖活力、克隆形成数、迁移和侵袭数、IL-8 分泌水平、N-cadherin阳性率及其蛋白表达、 TLR4、NF-κB、HMGB1、RAGE、cyclin D1、CDK2蛋白表达均显著降低（均 P < 0.05），细胞凋亡率、E-cadherin阳性率及其蛋白 表达、BAX/Bcl-2比值均显著升高（均P < 0.05）；同时过表达HMGB1则可逆转PHN对U251细胞增殖、迁移、侵袭及EMT的抑制 作用和对凋亡的促进作用（均P < 0.05）。结论：PHN通过HMGB1/RAGE信号通路抑制胶质瘤U251细胞增殖、侵袭及EMT进程。

[Abstract] Objective: To investigate the effects of phillyrin (PHN) on the proliferation, invasion, and epithelial-mesenchymal transition (EMT) of glioma U251 cells by adjusting the high mobility group protein B1 (HMGB1)/receptor of advanced glycation endproduct (RAGE) signaling pathway. Methods: Human glioma U251cells were assigned into the PHN-0 group (treated with 0 μmol/L PHN), the low, medium, and high-dose PHN groups (PHN-50、PHN-100、PHN-200 groups, treated with 50, 100, and 200 μmol/L PHN respectively), the PHN + pcDNA-NC group (treated with 200 μmol/L PHN after transfection of pcDNA-NC plasmid), and the PHN + HMGB1 group (treated with 200 μmol/L PHN after transfection of overexpressed HMGB1 plasmid). The proliferation ability of cells in each group was detected by the CCK-8 method and the clone formation assay. The apoptosis level of cells in each group was detected by flow cytometry. The migration and invasion abilities of cells in each group were detected by the Transwell assay. ELISA was used to detect the IL-8 secretion level of cells in each group. Immunofluorescence was used to detect the positive rates of N-cadherin and E-cadherin in cells of each group. WB assay was performed to detect the expression levels of Toll like receptor 4 (TLR4), nuclear factor-kappa B (NF- κB), HMGB1, RAGE, N-cadherin, E-cadherin, cell cycle protein D1 (cyclin D1), cyclin dependent kinase 2 (CDK2), B-lymphoblastoma-2 (Bcl-2), Bcl-2 associated X protein (BAX) proteins in cells of each group. Results: Compared with those in the PHN-0 group, the proliferation activity, the number of clone formation, the numbers of invasion and migration, IL-8 secretion levels, the positive rate and protein expression of N-cadherin, and the expressions of TLR4, NF-κB, HMGB1, RAGE, cyclin D1 and CDK2 protein in the PHN-50, PHN-100, and PHN-200 groups decreased significantly (all P < 0.05); and the apoptosis rate, the positivity rate and protein expression of E-cadherin , and the BAX/Bcl-2 ratio increased significantly (all P < 0.05). At the same time, overexpression of HMGB1 could reverse the inhibitory effects of PHN on the proliferation, migration, invasion and EMT of U251 cells, as well as its promoting effect on the apoptosis (all P < 0.05). Conclusion: PHN inhibits the proliferation, invasion and EMT progression of glioma U251 cells through the HMGB1/RAGE signaling pathway.

[基金项目] 2023年张家口市市级科技计划项目（No. 2322180D）