| 摘要: |
| 目的：观察牛膝总皂苷（TSA）体外干预人膝骨关节炎软骨细胞增殖及Ⅱ型胶原（CollagenⅡ）表达水平及最佳浓度。方法：将人膝骨关节炎软骨细胞进行分离、培养、鉴定,并将其分为6组,分别为0、0.01、0.05、0.1、0.5、1 mg/ml组。制备TSA存储液及6组不同TSA浓度DMEM/F12（D/F12）培养基,分别用6组培养基对P3代软骨细胞培养并进行细胞形态学观察；采用CCK-8法检测软骨细胞P1、P2、P3传代增殖情况,观察其生长曲线；采用Western-blot法检测P3代软骨细胞Collagen II蛋白表达、RT-PCR法检测CollagenⅡ mRNA表达。结果：软骨细胞体外分离并培养成功,经甲苯胺蓝染色鉴定细胞为软骨细胞来源；CCK-8法绘制的生长曲线符合Logistic生长曲线规律,P3代软骨细胞增殖速度高于P1、P2代；Western-Blot法检测结果显示,Collagen II蛋白表达量浓度为0.5 mg/ml时达最高,RT-PCR法检测结果显示,各组浓度AST能促进软骨组织Ⅱ型胶原mRNA表达上调,以0.5 mg/ml组最明显（P<0.05）。结论：TSA能体外有效促进软骨细胞增殖和Ⅱ型胶原mRNA表达上调,以0.5 mg/ml浓度最明显,具有促进软骨修复的作用,具体作用机制有待进一步研究。 |
| 关键词: 膝骨关节炎 软骨细胞 牛膝总皂苷 体外干预 实验研究 |
| DOI： |
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| In vitro intervention and optimal concentration of total saponins from Achyranthis Bidentatae Radix on the expression of type II collagen in chondrocytes of human knee osteoarthritis |
| CHEN Haoxiong,HUO Shaochuan,MA Dujun |
| (Shenzhen Hospital of Guangzhou University of Chinese Medicine(Futian),Shenzhen 518034,Guangdong,China;Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen 518033,Guangdong,China) |
| Abstract: |
| Objective：To investigate the in vitro intervention and optimal concentration of total saponins from Achyranthis Bidentatae Radix (TSA) on the expression of type II collagen (Collagen II) in chondrocytes of human knee osteoarthritis.Methods：Chondrocytes of human knee osteoarthritis were isolated,cultured,identified,and then divided into 0,0.01,0.05,0.1,0.5,and 1 mg/ml groups.TSA stock solution and DMEM/F12 medium with the above six concentrations of TSA were prepared for the culture of P3 chondrocytes,and cell morphology was observed；CCK-8 assay was used to observe the passage and proliferation of P1,P2,and P3 chondrocytes and their growth curves；Western blot was used to measure the protein expression of collagen II in P3 chondrocytes,and RT-PCR was used to measure the mRNA expression of collagen II.Results：Chondrocytes were successfully isolated and cultured in vitro,and the origin of cells was identified as chondrocytes by toluidine blue staining.The growth curve plotted based on CCK-8 assay was consistent with the logistic growth curve,and P3 chondrocytes had a higher proliferation rate than P1 and P2 chondrocytes.Western blot showed the highest protein expression of collagen II at the concentration of 0.5 mg/ml,and RT-PCR showed that TSA at each concentration promoted the upregulation of the mRNA expression of collagen II,with the most significant effect at the concentration of 0.5 mg/ml (P<0.05).Conclusion：TSA in vitro can effectively promote the proliferation of chondrocytes and the upregulation of the mRNA expression of collagen II,with the most significant effect at the concentration of 0.5 mg/ml,and it can also promote cartilage repair.Further studies are needed to clarify its mechanism of action. |
| Key words: knee osteoarthritis chondrocyte total saponins from Achyranthis Bidentatae Radix in vitro intervention experimental study |
