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Effects of rutin on osteoblast MC3T3-E1 differentiation, ALP activity and Runx2 protein expression
European Journal of Histochemistry ( IF 2 ) Pub Date : 2021-01-20 , DOI: 10.4081/ejh.2021.3195
Xin-Wei Liu 1 , Bin Ma 2 , Ying Zi 3 , Liang-Bi Xiang 1 , Tian-Yu Han 1
Affiliation  

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.



中文翻译:

芦丁对成骨细胞MC3T3-E1分化、ALP活性及Runx2蛋白表达的影响

芦丁作为一种黄酮类化合物,已被发现具有广泛的生物学功能,如抵抗炎症和氧化,预防脑出血和高血压。近年来发现它在骨质疏松症和其他骨科疾病中发挥重要作用。MC3T3-E1细胞随机分为对照组、rutin-1组(0.01 mmol/L)、rutin-2组(0.05 mmol/L)和rutin-3组(0.1 mmol/L)。细胞的成骨分化由成骨诱导液诱导。对照组给予最大剂量的药物溶剂。2~3天后,用含有芦丁的新鲜成骨诱导液代替溶剂。常规培养一定时间后,收集细胞用于后续实验。Runx2的表达Real-time PCR检测各组细胞基因;Western blot和免疫细胞化学检测Runx2蛋白的表达;ALP活性采用试剂盒法检测;通过茜素红染色分析成骨分化。Real-time PCR结果显示,与对照组相比,芦丁处理细胞可显着增加Runx2基因的表达(p<0.05);浓度越高,Runx2的表达越高基因,不同浓度组间差异显着(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。药物治疗组ALP活性显着高于对照组(p<0.05);使用不同浓度的组之间存在显着差异(p<0.05)。茜素红染色结果显示,各组均形成钙化结节,芦丁处理组形成的钙化结节面积大于对照组;浓度越大,面积越大。芦丁能促进成骨细胞分化;而且浓度越大,效果越好。

更新日期:2021-01-20
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