| 沈晓娟,程磊,刘琳.苍术素调节腺苷酸活化蛋白激酶 /沉默信息调节因子 1信号通路对白细胞介素 -1β诱导软骨细胞自噬和凋亡的影响[J].安徽医药,2024,28(5):899-903. |
| 苍术素调节腺苷酸活化蛋白激酶 /沉默信息调节因子 1信号通路对白细胞介素 -1β诱导软骨细胞自噬和凋亡的影响 |
| Effect of atractylodin on IL-1β-induced autophagy and apoptosis of chondrocytes by regulating AMPK/SIRT1 signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2024.05.011 |
| 中文关键词: 苍术属 白细胞介素 1β 软骨细胞 AMPK/SIRT1信号通路 自噬 |
| 英文关键词: Atractylodes Interleukin-1β Chondrocytes AMPK/SIRT1 signal pathway Autophagy |
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| 中文摘要: |
| 目的探讨苍术素(ATR)通过调节腺苷酸活化蛋白激酶 /沉默信息调节因子 1(AMPK/SIRT1)信号通路对白细胞介素 1β(IL-1β)诱导的乳鼠软骨细胞自噬与凋亡的影响。方法 2022年 1―8月从乳鼠中分离软骨细胞并鉴定；使用不同浓度苍术素(0、5.0、10.0、20.0、40.0、80.0 μmol/L)预处理软骨细胞，再使用 IL-1β诱导软骨细胞培养 24 h，噻唑蓝( MTT)法检测增殖活性，筛选最佳药物浓度；将软骨细胞分为对照组、 IL-1β组( 10 μg/L IL-1β)、苍术素组( 10 μg/L IL-1β+20μmol/L苍术素)、苍术素 +AMPK抑制剂组(苍术素 +compound C组， 10 μg/L IL-1β+20 μmol/L苍术素 +50 μmol/L compound C)；流式细胞术与 TUNEL法检测软骨细胞凋亡，单丹磺酰尸胺( MDC)染色观察自噬小体；蛋白质印迹法检测自噬及 AMPK/SIRT1通路相关蛋白表达。结果与对照组比较， IL-1β组软骨细胞增殖活性、自噬水平、微管相关蛋白轻链 3Ⅱ/Ⅰ(LC3Ⅱ/LC3Ⅰ)［( 0.47±0.08)比( 1.23±0.14)］、 Beclin-1［( 0.67±0.09)比( 1.45±0.19)］、自噬相关基因 -5(Atg5)［( 0.35±0.06)比( 1.14±0.13)］、磷酸化 AMPK(p-AMPK)［(0.37±0.05)比( 0.91±0.05)］、SIRT1［(0.51±0.06)比( 1.31±0.14)］磷酸化 Unc-51样自噬激活蛋白酶(p-ULK1)蛋白表达［(0.25±0.04)比(0.85±0.11)］降低，细胞凋亡率［(28.46±3.55)%比( 2.54±0.82)%］升高( P<0.05)；与 IL-1β组比较，苍术素组细胞增殖活性、自噬水平、 LC3Ⅱ/LC3Ⅰ、Beclin-1、Atg5、p-AMPK、SIRT1、p-ULK1蛋白表达升高，细胞凋亡率降低( P<0.05)； compound C可逆转苍术素对 IL-1β诱导的软骨细胞自噬激活作用。结论苍术素通过激活 AMPK/SIRT1信号通路促进 IL-1β诱导的大鼠软骨细胞自噬，抑制细胞凋亡。 |
| 英文摘要: |
| Objective To investigate the impacts of atractylodin (ATR) on interleukin-1β (IL-1β) induced autophagy and apoptosis of neonatal rat chondrocytes by regulating AMP-activated protein kinase/silencing Message regulator 1(AMPK/SIRT1) signaling pathway. Methods Chondrocytes were isolated from suckling mice and identified from January to August 2022; different concentrations ofatractylodin (0, 5.0, 10.0, 20.0, 40.0, 80.0 μmol/L) was used to pretreat chondrocytes, and then IL-1β was used to induce chondrocytesto culture for 24 hours, MTT assay was used to detect the proliferation activity to screen the best drug concentration; chondrocytes weredivided into Control group, IL-1β group (10 μg/L IL-1β), ATR group (10 μg/L IL-1β +20 μmol/L ATR), and ATR+AMPK inhibitor group (ATR+compound C group, 10 μg/L IL-1β+20μmol/L ATR+50μmol/L compound C), flow cytometry and TUNEL were used to de-tect chondrocyte apoptosis, the autophagosomes were observed by MDC staining; Western blotting was used to detect the expression ofautophagy and AMPK/SIRT1 pathway related proteins.Results Compared with the Control group, the proliferation activity of chondro-cytes, autophagy level, protein expression of Microtubule-associated protein light chain 3Ⅱ/Ⅰ(LC3 Ⅱ/LC3 Ⅰ) [(0.47±0.08) vs. (1.23± 0.14)], Beclin-1 [(0.67±0.09) vs. (1.45±0.19)], Autophagy-related gene(Atg5) [(0.35±0.06) vs. (1.14±0.13)], Phosphorylated AMPK(p-AMPK) [(0.37±0.05) vs. (0.91±0.05)], SIRT1 [(0.51±0.06) vs. (1.31±0.14)], andPhosphorylated Autophagy Activating Kinase1(p-ULK1) [(0.25±0.04) vs. (0.85±0.11)] in IL-1β group decreased, and the apoptosis rate [(28.46±3.55)% vs. (2.54±0.82)%] increased (P<0.05); compared with IL-1β group, the proliferation activity of chondrocytes, autophagy level, protein expression of LC3 Ⅱ/LC3 Ⅰ, Beclin-1, Atg5, p-AMPK, SIRT1, and p-ULK1 in ATR group increased, and the apoptosis rate decreased (P<0.05); compound C was able to re-verse the activation of IL-1β-induced chondrocyte autophagy induced by atractylonin.Conclusion Atractylodin promotes IL-1β-in-duced autophagy of rat chondrocytes and inhibits apoptosis by activating AMPK/SIRT1 signaling pathway. |
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