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陈伟达,石玮,杨继国,陈腾.三七总皂苷经miR-24靶向C-myc对骨关节炎大鼠软骨细胞凋亡与增殖的影响[J].浙江中西医结合杂志,2021,31(4):
三七总皂苷经miR-24靶向C-myc对骨关节炎大鼠软骨细胞凋亡与增殖的影响
Effect of Panax Notoginseng Saponins targeting c-myc via miR-24 on chondrocyte apoptosis and proliferation in osteoarthritis rats
投稿时间:2020-07-07  修订日期:2021-03-16
DOI:
中文关键词:  三七总皂苷  miR-24  C-myc  骨关节炎  软骨细胞  凋亡  增殖
英文关键词:Total saponins of Panax notoginseng  miR-24  C-myc  Osteoarthritis  Chondrocytes  Apoptosis  Proliferation
基金项目:
作者单位E-mail
陈伟达 台州市第一人民医院手足外科 daweichen847@163.com 
石玮 台州市第一人民医院手足外科  
杨继国 台州市第一人民医院手足外科  
陈腾 台州市第一人民医院手足外科  
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中文摘要:
      目的 探讨三七总皂苷经微小RNA-24(miR-24)靶向原癌基因(C-myc)对骨关节炎大鼠软骨细胞凋亡与增殖的影响。方法 木瓜蛋白酶水溶液注射至大鼠膝关节腔内诱导骨关节炎,分离两侧膝关节骨关节炎软骨;设立骨关节炎软骨细胞组、三七总皂苷低、中、高剂量组(三七总皂苷终浓度分别为:100、200、400μg/ml),于37℃、5%CO2、20% O2培养72h,培养结束后,CCK-8试剂盒测定细胞活力、吉姆萨染色测定菌落水平、流式细胞仪测定细胞凋亡水平、实时荧光定量PCR(qRT-PCR)法与蛋白免疫印迹(WB)法测定miR-24、C-myc mRNA及C-myc蛋白水平。结果 与骨关节炎软骨细胞组比较,三七总皂苷低、中、高剂量组OD值、存活率、细胞菌落数、细胞miR-24 mRNA水平升高(P<0.05),凋亡率、C-myc mRNA和蛋白水平降低(P<0.05);且随着三七总皂苷剂量的增加,三七总皂苷各剂量组OD值、存活率、细胞菌落数、细胞miR-24 mRNA水平逐渐升高(P<0.05),凋亡率、C-myc mRNA和蛋白水平逐渐降低(P<0.05),呈剂量依赖性(P<0.05)。结论 三七总皂苷能抑制骨关节炎大鼠软骨细胞凋亡,促进骨关节炎大鼠软骨细胞增殖;其机制可能与三七总皂苷能靶向上调miR-24 mRNA进而抑制C-myc mRNA和蛋白的表达有关。
英文摘要:
      Objective: To investigate the effect of Panax Notoginseng Saponins targeting C-myc via MicroRNA-24(miR-24) on chondrocyte apoptosis and proliferation in osteoarthritis rats. Methods: Papain aqueous solution was injected into the knee joint cavity of rats to induce osteoarthritis and to separate the osteoarthritis cartilage on both sides of the knee joint. The Osteoarthritis Chondrocytes group and the low, medium and high dose group of total saponins of Panax notoginseng (the final concentration of total saponins of Panax notoginseng: 100, 200 and 400 μg/ml) were set up and cultured at 37℃, 5%CO2 and 20%O2 for 72h. After the culture, the cell viability were measured by CCK-8 kit, the colony level was determined by Giemsa staining, Cell apoptosis was measured by flow cytometry, The mRNA and protein levels of mir-24, C-myc and C-myc protein were measured by real-time quantitative PCR (qRT-PCR) and Western blot (WB). Results: Compared with Osteoarthritis Chondrocytes group, the OD value, survival rate, cell colony number, level of mir-24 mRNA of total saponins of Panax notoginseng in low, medium and high dose groups were increased (P<0.05); the apoptosis rate, C-myc mRNA and protein in cells were decreased (P<0.05). And with the increase of Panax notoginseng saponins dose, the OD value, survival rate, cell colony number, level of mir-24 mRNA of total saponins of the each dose group of total saponins of notoginseng were gradual increase (P<0.05); the apoptosis rate, C-myc mRNA and protein in cells were gradual decreased (P<0.05); it was dose-dependent (P<0.05). Conclusion: Total saponins of Panax notoginseng can inhibit the apoptosis of chondrocytes in osteoarthritis rats and promote the proliferation of chondrocytes in osteoarthritis rats. The mechanism may be related to the fact that total saponins of Panax notoginseng can target up-regulated miR-24 mRNA and then inhibit the expression of c-myc mRNA and protein.
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