Department of Orthopaedics, General Hospital of Northern Theater Command, Shenyang, Liaoning.
Department of Orthopaedics, Tongji Hospital, Tongji University School of Medicine, Shanghai.
Eur J Histochem. 2021 Jan 20;65(1):3195. doi: 10.4081/ejh.2021.3195.
As a flavonoid, rutin has been found to have a wide range of biological functions, such as resisting inflammation and oxidation, and preventing cerebral hemorrhage and hypertension. It has been found to play an important role in osteoporosis and other orthopedic diseases in recent years. MC3T3-E1 cells were randomly divided into a control group, a rutin-1 group (0.01 mmol/L), a rutin-2 group (0.05 mmol/L) and a rutin-3 group (0.1 mmol/L). Osteogenic differentiation of cells was induced by osteogenic induction fluid. The control group was treated with the maximum dose of drug solvent. 2~3 days later, the solvent was replaced with fresh osteogenic induction fluid containing rutin. After a certain period of routine culture, the cells were collected for subsequent experiments. The expression of Runx2 gene in cells in all groups was detected by Real-time PCR; the expression of Runx2 protein was detected by Western blot and immunocytochemistry; the activity of ALP was detected by reagent kit method; osteogenic differentiation was analyzed by alizarin red staining. The results of Real-time PCR showed that, compared with the control group, the treatment of cells with rutin can significantly increase the expression of Runx2 gene (p<0.05); the higher the concentration, the higher the expression of Runx2 gene, and significant differences were found among groups in which different concentrations were used (p<0.05); the results of Western blot and IHC showed that the expression trend of Runx2 protein in each group was consistent with PCR results. In drug treatment groups, the activity of ALP was significantly higher than that in the control group (p<0.05); there were significant differences among groups in which different concentrations were used (p<0.05). The results of alizarin red staining showed that calcified nodules were formed in all groups and that the area of calcified nodules formed in groups treated with rutin was greater than that in the control group; the greater the concentration, the larger the area. Rutin can promote osteoblastic differentiation; and the greater the concentration, the more effective it is.
芦丁作为一种类黄酮，已被发现具有广泛的生物学功能，如抗炎和抗氧化，预防脑出血和高血压。近年来，它在骨质疏松症和其他骨科疾病中被发现起着重要作用。MC3T3-E1 细胞被随机分为对照组、芦丁-1 组（0.01mmol/L）、芦丁-2 组（0.05mmol/L）和芦丁-3 组（0.1mmol/L）。用成骨诱导液诱导细胞成骨分化。对照组用最大剂量的药物溶剂处理。2~3 天后，用含芦丁的新鲜成骨诱导液替换溶剂。经过一定时间的常规培养，收集细胞进行后续实验。用 Real-time PCR 检测各组细胞中 Runx2 基因的表达；用 Western blot 和免疫细胞化学检测 Runx2 蛋白的表达；用试剂盒法检测 ALP 的活性；用茜素红染色分析成骨分化。Real-time PCR 结果显示，与对照组相比，芦丁处理细胞可显著增加 Runx2 基因的表达（p<0.05）；浓度越高，Runx2 基因表达越高，不同浓度组间差异有统计学意义（p<0.05）；Western blot 和 IHC 结果显示，各组 Runx2 蛋白表达趋势与 PCR 结果一致。药物处理组 ALP 活性明显高于对照组（p<0.05）；不同浓度组间差异有统计学意义（p<0.05）。茜素红染色结果显示，各组均形成钙化结节，芦丁处理组钙化结节形成面积大于对照组；浓度越高，面积越大。芦丁能促进成骨细胞分化；且浓度越大，效果越显著。
