摘要
目的:研究不同浓度的白芍总苷在不同时间干预下对人膝关节骨性关节炎滑膜成纤维细胞增殖活性的影响,探讨时间与浓度的变化规律和对滑膜炎症反应的作用。方法:膝关节置换术中取得新鲜滑膜组织经胶原酶法分离培养,于倒置显微镜下观察细胞形态;将白芍总苷以1mg/m L为初始浓度,4倍稀释成11个浓度依次作用于滑膜成纤维细胞,用四甲基偶氮唑盐法分别测定不同浓度的白芍总苷在24、48、72h干预后对滑膜成纤维细胞增殖水平的影响,并计算生长抑制率和存活分数。结果:经24、48、72h的干预后,低浓度的白芍总苷对人膝关节骨性关节炎滑膜成纤维细胞生长有轻度促进作用,中高浓度(>15.625μg/m L)则有显著的抑制作用(P<0.05),且呈时间和浓度依赖性。结论:四甲基偶氮唑盐实验可用于评价白芍总苷对滑膜成纤维细胞增殖活性的影响;测定并计算各生长抑制率和存活分数,探讨中药有效成分与细胞增殖活性的关系,为进一步明确其药理作用机制、建立相应滑膜炎症反应细胞模型、探寻针对骨性关节炎各环节的靶向治疗提供实验参数。
Objective： To investigate influence of total glucosides of paeonia（TGP） on proliferation of fibroblast-like synovicyte（FLS） in patients with knee osteoarthritis（knee OA）, as well as to study the change rules and the effects on synovitis of time and concentration. Methods： FLS were obtained and cultured from the patients with knee OA in joint replacement, and observed the morphology with inverted microscopy. FLS were treated with 11 concentrations of TGP（ the initial was 1mg/mL by 4 times dilution）, and the cell proliferation of 24/48/72 h was examined by methyl thiazolyl tetrazolium assay. The survival fraction（SF） and inhibition rate（IR） were calculated. Results： After 24/48/72 h low concentration of TGP showed a slight promotion action on FLS, while middle and high concentration（15.625μg/mL） could significantly inhibit the proliferation of FLS（P〈0.05） in time and concentration-dependent manners. Conclusion： MTT assay can be used in test of the proliferation of FLS with TGP, as well as determination of IR and SF, which could be used for further study as a cell model for synovitis. To explore the relationship between TCM effective constituents and cellular proliferative activity, and guide explicit pharmacological mechanism, as well as further study for the target therapy in each link of knee OA.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2016年第11期4853-4856,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金面上项目(No.81373662)
北京中医药大学自主选题项目(No.2015-JYBZZ-XS)~~
