周哲人,马 训,么焕开,吴奇憧,张心怡,叶子欣,杨 锋.柚皮苷和木犀草素联合应用对大鼠BMSCs诱导成骨过程中Wnt/β-cantein通路相关基因表达的影响[J].现代生物医学进展英文版,2023,(2):258-262. |
柚皮苷和木犀草素联合应用对大鼠BMSCs诱导成骨过程中Wnt/β-cantein通路相关基因表达的影响 |
Combined Application of Naringin and Luteolin on the Osteogenesis of Rat BMSCs and Related Genes Expression of Wnt/β-cantein Pathway |
Received:April 23, 2022 Revised:May 18, 2022 |
DOI:10.13241/j.cnki.pmb.2023.02.010 |
中文关键词: 柚皮苷 木犀草素 骨髓间充质干细胞 成骨 Wnt/β-cantein通路 |
英文关键词: Naringin Luteolin Bone marrow mesenchymal stem cells Osteogenesis Wnt/β- cantein pathway |
基金项目:江苏省高等学校自然科学研究面上项目 (20KJD320006);徐州市科技计划项目(KC21197);江苏省大学生创新训练项目 (202110313064Y) |
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中文摘要: |
摘要 目的:探讨柚皮苷和木犀草素联合应用对大鼠骨髓间充质干细胞(BMSCs)诱导成骨过程中Wnt/β-cantein通路相关基因表达的影响。方法:取大鼠股骨中提取的BMSCs,分别建立A组(空白组)、B组给予柚皮苷溶液10 μmol/L,C组给予木犀草素溶液5 μmol/L;D组给予骨碎补总黄酮溶液10 mg/mL;E组给予柚皮苷-木犀草素混合溶液配伍比为10 μmol/L:5 μmol/L,并诱导其向成骨细胞分化。应用碱性磷酸酶(ALP)染色及分光光度法检测第7 d各组细胞ALP活性。应用实时荧光定量聚合酶链反应(qRT-PCR)检测第7 d各组细胞Wnt/β-cantein通路相关基因及成骨基因的表达。结果:B组、C组、D组、E组细胞562 nm波长下光密度(OD)值显著高于A组,E组细胞562 nm波长下OD值最高,显著高于B组、C组、D组(P<0.05)。B组、C组、D组、E组细胞ALP、骨钙素(OCN)、Runt相关转录因子2(RUNX2)基因表达水平显著高于A组,E组ALP、OCN、RUNX2基因表达水平最高,显著高于B组、C组、D组(P<0.05)。B组、C组、D组、E组细胞β-catenin、Cyclin D1基因表达水平显著高于A组;B组、E组LEF-1基因表达水平显著高于A组;E组β-catenin、LEF-1、Cyclin D1基因表达水平最高,显著高于B组、C组、D组(P<0.05)。结论:柚皮苷和木犀草素均具有促进大鼠BMSCs增殖和诱导其成骨向分化的作用,柚皮苷和木犀草素联合应用诱导大鼠BMSCs增成骨作用最强,其主要机制与Wnt/β-cantein通路激活有关。 |
英文摘要: |
ABSTRACT Objective: To investigate the effect of naringin and luteolin on the osteogenesis of rat bone marrow mesenchymal stem cells (BMSCs) and Wnt/β-cantein pathway expression. Methods: BMSCs extracted from rat femur were established. Group A (blank group) and group B were given naringin solution for 10 μmol/L, group C was given luteolin solution for 5 μmol/L, group D was given Rhizoma Drynariae total flavonoids solution 10 mg/mL, group E was given naringin luteolin mixed solution with a compatibility ratio of 10 μmol/L: 5 μmol/L, and induce it to differentiate into osteoblasts. Alkaline phosphatase (ALP) staining and spectrophotometry were used to detect the ALP activity of cells in each group on the 7th day. Wnt/β-cantein pathway related genes and osteogenic genes was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Results: The OD value of cells in group B, group C, group D and group E at 562 nm wavelength was significantly higher than that in group A. The OD value of cells in Group E at 562 nm wavelength was the highest, which was significantly higher than that in group B, group C and group D (P<0.05). The expression levels of ALP, osteocalcin(OCN) and Runt related transcription factor 2(RUNX2) genes in group B, C, D and E were significantly higher than those in group A. the expression levels of ALP, OCN and RUNX2 genes in Group E were the highest, which were significantly higher than those in group B, C and D (P<0.05). β-catenin, Cyclin D1 in group B, group C, group D and group E were significantly higher than those in group A. The expression level of LEF-1 gene in group B and group E was significantly higher than that in group A. The gene expression levels of β-catenin, LEF-1, Cyclin D1 were the highest, which were significantly higher than those in group B, group C and group D(P<0.05). Conclusion: Naringin and luteolin can promote the proliferation and osteogenic differentiation of rat BMSCs. The combined application of naringin and luteolin has the strongest osteogenic effect on rat BMSCs, and its main mechanism is related to Wnt/ β- cantein activation pathway. |
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