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顾龙龙, 单妍, 梁璇, 李荣, 谢薇.三七皂苷R1通过激活AMPK/mTOR/ULK1信号通路促进脂多糖诱导的牙周炎细胞模型自噬[J].湖南中医药大学学报英文版,2025,45(11):2090-2097.[Click to copy
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| 三七皂苷R1通过激活AMPK/mTOR/ULK1信号通路促进脂多糖诱导的牙周炎细胞模型自噬 |
| 顾龙龙,单妍,梁璇,李荣,谢薇 |
| (昆明医科大学海源学院生物化学分子生物学教研室, 云南 昆明 651700) |
| 摘要: |
| 目的 基于5'-磷酸腺苷活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）/UNC-51样激酶1(ULK1)信号通路，探究三七皂苷R1对脂多糖（LPS）诱导的牙周炎细胞自噬的影响。方法 采用100 ng/mL LPS处理人牙周膜成纤维细胞（hPDLFs）24 h以构建牙周炎细胞模型。将牙周炎细胞分为control组、LPS组、LPS+三七皂苷R1组、LPS+三七皂苷R1+AMPK抑制剂Compound C组。流式细胞术检测细胞的凋亡情况，Western blot检测细胞中Bcl-2、Bax、cleaved Caspase-3、Beclin-1、p62、pAMPK、p-mTOR、p-ULK1蛋白表达水平及LC3-Ⅱ/Ⅰ蛋白表达比值，免疫荧光染色实验检测细胞中LC3蛋白的表达水平。结果 相对于control组，LPS组细胞的凋亡率升高（P<0.05），细胞中Bcl-2、Beclin-1、p-AMPK、p-ULK1蛋白表达水平及LC3-Ⅱ/Ⅰ比值降低（P<0.05），细胞中Bax、cleaved Caspase-3、p62、p-mTOR蛋白表达水平升高（P<0.05）；相对于LPS组，LPS+三七皂苷R1组、LPS+三七皂苷R1+Compound C组细胞的凋亡率降低（P<0.05），细胞中Bcl-2、Beclin-1、p-AMPK、p-ULK1、蛋白表达水平及LC3-Ⅱ/Ⅰ比值升高（P<0.05），细胞中Bax、cleaved Caspase-3、p62、p-mTOR降低（P<0.05）；相对于LPS+三七皂苷R1组，LPS+三七皂苷R1+Compound C组细胞的凋亡率升高（P<0.05），细胞中Bcl-2、Beclin-1、p-AMPK、p-ULK1蛋白表达水平及LC3-Ⅱ/Ⅰ比值降低（P<0.05），细胞中Bax、cleaved Caspase-3、p62、p-mTOR升高（P<0.05）。结论 三七皂苷R1能够促进LPS诱导的牙周炎细胞自噬并抑制其凋亡，其机制可能与激活AMPK/mTOR/ULK1信号通路有关。 |
| 关键词: 牙周炎 人牙周膜成纤维细胞 三七皂苷R1 AMPK/mTOR/ULK1信号通路 自噬 凋亡 |
| DOI：10.3969/j.issn.1674-070X.2025.11.009 |
| Received:July 23, 2025 |
| 基金项目:昆明医科大学海源学院2024年科学研究基金项目(2024HY013)。 |
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| Panax notoginseng saponin R1 activates the AMPK/mTOR/ULK1 signaling pathway to promote autophagy in a lipopolysaccharide-induced periodontitis cell model |
| GU Longlong, SHAN Yan, LIANG Xuan, LI Rong, XIE Wei |
| (Department of Biochemistry and Molecular Biology, Haiyuan College, Kunming Medical University, Kunming, Yunnan 651700, China) |
| Abstract: |
| Objective To investigate the effects of Panax notoginseng saponin R1（NGR1） on autophagy in lipopolysaccharide（LPS）-induced periodontitis cells via the adenosine 5’-monophosphate-activated protein kinase（AMPK）/mammalian rapamycin target protein（mTOR）/Unc-51 like kinase 1（ULK1） signaling pathway. Methods Human periodontal ligament fibroblasts（hPDLFs） were treated with 100 ng/mL LPS for 24 h to establish a periodontitis cell model. The periodontitis cells were divided into control group,LPS group, LPS+NGR1 group, and LPS+NGR1+AMPK inhibitor Compound C group. Flow cytometry was used to check apoptosis of the cells. The protein expression levels of Bcl-2, Bax, cleaved Caspase-3, Beclin-1, p62, p-AMPK, p-m TOR, and p-ULK1, as well as the LC3-Ⅱ/Ⅰ ratio, were determined by Western blot. Immunofluorescence staining assay was used to examine the protein expression level of LC3. Results Compared with the control group, the LPS group exhibited a significant increase in the apoptosis rate of cells（P＜0.05）. The protein expression levels of Bcl-2, Beclin-1, p-AMPK and p-ULK1 in cells, as well as the LC3-Ⅱ/Ⅰ ratio in cells decreased（P＜0.05）. Conversely, the protein expression levels of Bax, cleaved Caspase-3, p62, and p-mTOR in cells were elevated（P＜0.05）. Compared with the LPS group, the apoptosis rate of cells in LPS+Panax notoginseng saponin R1 group and LPS+Panax notoginseng saponin R1+Compound C group decreased（P＜0.05）. The protein expression levels of Bcl-2, Beclin-1, p-AMPK,and p-ULK1, as well as the LC3-Ⅱ/Ⅰ ratio, significantly increased（P ＜0.05）. Conversely, the protein expression levels of Bax,cleaved Caspase-3, p62, and p-mTOR significantly decreased（P＜0.05）. Compared with the LPS+notoginsenoside R1 group, the cells in the LPS+notoginsenoside R1+Compound C group exhibited an increased apoptosis rate（P＜0.05）, along with decreased levels of Bcl-2,Beclin-1, the ratio of LC3-Ⅱ/Ⅰ, p-AMPK, and p-ULK1（P＜0.05）, while showing increased levels of Bax, cleaved Caspase-3, p62, and p-mTOR（P＜0.05）. Conclusion Panax notoginseng saponin R1 promotes autophagy and inhibits apoptosis of LPS-induced periodontitis cells, and its mechanism may be related to the activation of AMPK/mTOR/ULK1 signaling pathway. |
| Key words: periodontitis human periodontal ligament fibroblasts Panax notoginseng saponin R1 AMPK/mTOR/ULK1 signaling pathway autophagy apoptosis |
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