目的 探讨鸢尾素促进高糖环境中兔骨髓间充质干细胞(BMSCs)成骨分化的作用机制。方法 兔BMSCs随机分为对照组、高糖组、鸢尾素组(高糖+鸢尾素)和氯奎组(高糖+鸢尾素+氯奎)。成骨诱导培养5 d后,Western blot检测自噬相关蛋白P62和LC3B的表达,实时荧光定量反转录PCR(qRT-PCR)检测成骨相关基因Runx2、ALP、COL-I和OCN的转录表达,试剂盒检测碱性磷酸酶(ALP)活性。结果 与对照组相比,高糖组P62蛋白表达升高同时LC3B-II表达降低,下调Runx2、ALP和COL-I的转录表达并降低ALP活性(P＜0.05);鸢尾素干预降低P62蛋白水平同时升高LC3B-II的表达,上调Runx2、ALP和COL-I的转录表达并升高ALP活性(P＜0.05);当使用氯喹抑制自噬后,鸢尾素上调Runx2和ALP的转录表达并升高ALP活性的作用明显减弱(P＜0.05)。结论 鸢尾素通过激活自噬促进高糖环境中BMSCs成骨分化。
Abstract
Objective To explore irisin’s promotion mechanism of osteoblast differentiation of rabbit BMSCs in high glucose condition.Methods BMSCs of rabbit were divided into 4 groups: control, HG (high glucose), irisin (HG+irisin) and CQ (chloroquine; HG+irisin+CQ). After cultured in osteogenic differentiation medium for 5 days, western blotting was used to detect autophagy-related protein levels of P62 and LC3B; qRT-PCR was used to evaluate the expression levels of Runx2, ALP, COL-I and OCN; ALP activity was also tested.Results Compared with the control group, HG group significantly increased P62 and decreased LC3B-II protein levels, down-regulated mRNA expression of Runx2, ALP and COL-I, and reduced ALP activity (P＜0.05);however, irisin obviously decreased P62 and increased LC3B-II protein levels, up-regulated mRNA expression of Runx2, ALP and COL-I, and promoted ALP activity (P＜0.05); moreover, when inhibited autophagy with CQ, the effect of irisin was blunted (P＜0.05).Conclusion Irisin promotes osteoblast differentiation of BMSCs in high glucose condition via activating autophagy.
关键词
鸢尾素 /
自噬 /
骨髓间充质干细胞 /
成骨细胞
Key words
irisin /
autophagy /
BMSCs /
osteoblast
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