中国骨质疏松杂志

葛根素调控SIRT1-FOXO1通路抑制成骨细胞凋亡

Puerarin regulates SIRT1-FOXO1 pathway to inhibit osteoblast apoptosis

DOI：10.3969/j.issn.1006-7108.2025.05.008

中文关键词: 葛根素沉默信息调节因子1-叉头状转录因子O1 类固醇成骨细胞自噬

英文关键词:puerarin silent information regulator 1-forkhead box transcription factor O1 steroids osteoblasts autophagy

基金项目:武汉市医学科研项目（WZ21C09）

作者	单位
周凡1* 高扬1 胡艳平1 向超1 万骐 1 周茹2	1.武汉市中医医院骨科，湖北武汉 430000 2.武汉市中医医院脑病科，湖北武汉 430000

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中文摘要:

目的探讨葛根素（puerarin，PR）调节沉默信息调节因子1（SIRT1）-叉头状转录因子O1（FOXO1）信号通路对类固醇诱导的成骨细胞自噬和凋亡的影响。方法用浓度为0～50 μmol/L的PR及10 μmol/L的地塞米松（DEX）共同处理小鼠成骨细胞（MC3T3-E1），MTT法筛选最佳PR作用浓度；将MC3T3-E1细胞分为对照组（不进行任何干预）、DEX组（10 μmol/L DEX）、PR+DEX组（10 μmol/L DEX+40 μmol/L PR）、DEX+EX527组（10 μmol/L DEX+100 nmol/L的SIRT1抑制剂EX527）、PR+DEX+EX527组（10 μmol/L DEX+40 μmol/L PR+100 nmol/L EX527），MTT法、流式细胞术、MDC法测定各组细胞增殖活力、凋亡率及细胞自噬数量；Western Blot检测SIRT1-FOXO1通路相关蛋白、自噬标志蛋白Beclin-1、LC3及凋亡蛋白Bax、Bcl-2表达水平。结果 0～40 μmol/L的PR可促进DEX诱导的MC3T3-E1细胞增殖活力，40 μmol/L PR处理后MC3T3-E1细胞增殖活力最高，选择40 μmol/L PR进行后续实验。与对照组比较，DEX组MC3T3-E1细胞增殖活力、自噬阳性率、SIRT1、FOXO1、Beclin-1、LC3、Bax蛋白表达均降低，凋亡率及Bcl-2蛋白表达升高（P＜0.05）；与DEX组对比，PR+DEX组凋亡率及Bcl-2蛋白表达降低，上述其余指标均升高（P＜0.05），DEX+EX527组凋亡率及Bcl-2蛋白表达升高，上述其余指标均降低（P＜0.05）；与PR+DEX组对比，PR+DEX+EX527组凋亡率及Bcl-2蛋白表达升高，上述其余指标均降低（P＜0.05）。结论 PR可通过激活SIRT1-FOXO1信号通路增强MC3T3-E1细胞自噬进而抑制DEX诱导的细胞凋亡。

英文摘要:

Objective To investigate the effect of puerarin (PR) on steroid induced autophagy and apoptosis of osteoblasts by regulating the silent information regulator 1 (SIRT1) - forkhead box transcription factor O1 (FOXO1) signaling pathway. Methods Mouse osteoblasts (MC3T3-E1) were treated with 0-50 μmol/L and 10 μmol/L dexamethasone (DEX), and the optimal concentration of PR concentration was selected by MTT method; MC3T3-E1 cells were separated into a control group (without any intervention), a DEX group (10 μmol/L DEX), a PR+DEX group (40 μmol/L PR + 10 μmol/L DEX), a DEX+EX527 group (10 μmol/L DEX+100 nmol/LSIRT1 inhibitor EX527), and a PR+DEX+EX527 group (10 μmol/L DEX+40 μmol/L PR+100 nmol/L EX527), MTT assay, flow cytometry, and MDC assay were applied to determine the proliferation activity, apoptosis rate, and autophagy quantity of cells in each group; Western blot was applied to detect the expression levels of SIRT1-FOXO1 pathway related proteins, autophagy marker proteins Beclin-1, LC3, and apoptotic proteins Bax and Bcl-2. Results PR at 0-40 μmol/L can promote the proliferation activity of MC3T3-E1 cells induced by DEX, after treatment with 40 μmol/L PR, MC3T3-E1 cells showed the highest proliferation activity. 40 μmol/L PR was selected for subsequent experiments. Compared with the control group, the proliferation activity, autophagy positivity rate, SIRT1, FOXO1, Beclin-1, LC3, and Bax protein expression of MC3T3-E1 cells in the DEX group decreased, while the apoptosis rate and expression of Bcl-2 protein increased (P<0.05); compared with the DEX group, the PR+DEX group showed a decrease in apoptosis rate and Bcl-2 protein expression, while all other indicators mentioned above increased (P<0.05); compared with the PR+DEX group, the PR+DEX+EX527 group showed an increase in apoptosis rate and Bcl-2 protein expression, while all other indicators mentioned above decreased (P<0.05), the DEX+EX527 group showed an increase in apoptosis rate and Bcl-2 protein expression, while all other indicators mentioned above decreased (P<0.05). Conclusion PR can enhance autophagy in MC3T3-E1 cells and inhibit DEX induced apoptosis by activating the SIRT1-FOXO1 signaling pathway.

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