黄芩苷对脂多糖诱导的人牙周膜干细胞增殖、迁移作用及机制
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篇名: 黄芩苷对脂多糖诱导的人牙周膜干细胞增殖、迁移作用及机制
TITLE: Effect and mechanism of baicalin on the proliferation and migration of human periodontal ligament stem cells induced by lipopolysaccharide
摘要: 目的 探究黄芩苷对脂多糖(LPS)诱导的人牙周膜干细胞(hPDLSCs)增殖、迁移及Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号通路的调控作用。方法将hPDLSCs分为对照组、LPS组、黄芩苷不同浓度(0.1、1、10mg/L)组,采用ELISA法和CCK-8法测定细胞炎症因子[白细胞介素6(IL-6)、IL-1β和肿瘤坏死因子α(TNF-α)]含量和细胞活力,以筛选最适黄芩苷浓度并进行后续通路验证实验。随后将细胞分为对照组、LPS组、最适黄芩苷浓度组、抑制剂组(10μg/mLLPS+1mg/L黄芩苷+3μmol/LJAK2/STAT3通路抑制剂AG490),干预24h后检测hPDLSCs增殖率、凋亡率、迁移率、侵袭细胞数、细胞周期素D1(CyclinD1)、胱天蛋白酶3(caspase-3)mRNA和蛋白表达及JAK2/STAT3信号通路相关蛋白表达。结果根据细胞炎症因子含量和细胞活力结果选择1mg/L为黄芩苷最适浓度。与对照组比较,LPS组细胞增殖率、迁移率、侵袭细胞数、CyclinD1mRNA及蛋白表达水平显著降低,细胞凋亡率、caspase-3mRNA及蛋白表达、p-JAK2和p-STAT3蛋白表达显著升高(P<0.05);经1mg/L黄芩苷干预后,上述指标均得到显著改善(P<0.05);而抑制剂组上述指标的改善程度更加显著(P<0.05)。结论黄芩苷可通过抑制JAK2/STAT3信号通路来促进LPS诱导的hPDLSCs增殖、迁移和侵袭,抑制其凋亡及炎症反应。
ABSTRACT: OBJECTIVE To explore the regulatory effects of baicalin on the proliferation and migration of human periodontal ligament stem cells (hPDLSCs) induced by lipopolysaccharide (LPS) and Janus protein tyrosine kinase 2 (JAK2)/signal transduction and transcription activator 3 (STAT3) signaling pathways. METHODS hPDLSCs were divided into control group, LPS group, different concentration baicalin groups (0.1, 1 and 10 mg/L). ELISA method and CCK-8 assay were used to determine the contents of cell inflammatory factors [interleukin 6 (IL-6), IL-1β and tumor necrosis factor-α (TNF-α)] and cell viability, so as to screen the optimal concentration of baicalin for follow-up pathway validation experiments. The cells were then divided into control group, LPS group, optimal baicalin concentration group and inhibitor group (10 μg/mL LPS+1 mg/L baicalin +3 μmol/L JAK2/STAT3 pathway inhibitor AG490). After treated for 24 h, the proliferation rate of hPDLSCs, apoptosis rate, migration rate, invasion cell number, mRNA and protein expressions of Cyclin D1 and caspase-3, the expression of JAK2/STAT3 pathway-related proteins were all detected. RESULTS According to cell inflammatory factors and cell viability, 1 mg/L was selected as the optimal concentration of baicalin. Compared with control group, cell proliferation rate, migration rate, invasion cell number, Cyclin D1 mRNA and protein expression were significantly decreased in LPS group, while cell apoptosis rate, caspase-3 mRNA and protein expression, p-JAK2 and p-STAT3 protein expression were significantly increased (P<0.05). After treated with 1 mg/L baicalin, the above indexes were reversed significantly (P<0.05). The improvement of above indexes in the inhibitor group was more obvious (P<0.05). CONCLUSIONS Baicalin can promote the proliferation, migration and invasion of LPS-induced hPDLSCs and inhibit their apoptosis and inflammation by blocking the JAK2/STAT3 pathway.
期刊: 2023年第34卷第10期
作者: 王桥;冯博;高永志
AUTHORS: WANG Qiao,FENG Bo,GAO Yongzhi
关键字: 牙周炎;黄芩苷;牙周膜干细胞;Janus蛋白酪氨酸激酶2/信号转导和转录激活因子3信号通路;增殖;迁移
KEYWORDS: periodontitis; baicalin; periodontal ligament
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