牛膝总皂苷体外干预膝关节炎软骨细胞增殖与凋亡的研究  

Effects of total saponins of Achyranthes bidentata on proliferation and apoptosis of chondrocytes in vitro

作　　者：陈浩雄 徐启良 黄竞杰 林新珍 徐宁达 CHEN Haoxiong;XU Qiliang;HUANG Jingjie;LIN Xinzhen;XU Ningda(Shenzhen Hospital of Guangzhou University of Chinese Medicine(Futian),Shenzhen 518034,Guangdong,China)

机构地区：[1]广州中医药大学深圳医院(福田),广东深圳518034

出　　处：《现代中西医结合杂志》2021年第22期2400-2404,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：广东省中医药管理局资助项目(20191295)。

摘　　要：目的观察不同浓度牛膝总皂苷对人膝骨关节炎软骨细胞增殖、凋亡及炎性因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)表达的影响。方法实验分为空白组、牛膝总皂苷0.01 mg/mL组、牛膝总皂苷0.05 mg/mL组、牛膝总皂苷0.1 mg/mL组、牛膝总皂苷0.5 mg/mL组、牛膝总皂苷1 mg/mL组。取第3代人膝骨关节炎软骨细胞,空白组常规培养,牛膝总皂苷各组加入相应浓度牛膝总皂苷D/F12培养,采用CCK-8法及Annexin V-FITC/PI检测细胞增殖和凋亡情况,应用放射免疫分析法检测IL-1β水平,应用ELISA法检测TNF-α水平。结果牛膝总皂苷各组培养第3天细胞增殖达到高峰(空白组第5天达到高峰),之后呈下降趋势,牛膝总皂苷0.5 mg/mL组各时间点细胞增殖OD值均明显高于其他组(P均<0.05)。培养3 d后,牛膝总皂苷0.01 mg/mL和0.05 mg/mL组早期和晚期细胞凋亡率及总凋亡率与空白组比较差异均无统计学意义(P均>0.05);牛膝总皂苷0.1 mg/mL、0.5 mg/mL、1 mg/mL组早期和晚期细胞凋亡率及总凋亡率均明显低于空白组(P均<0.05),且牛膝总皂苷0.5 mg/mL组细胞总凋亡率明显低于牛膝总皂苷0.1 mg/mL和1 mg/mL组(P均<0.05)。干预72 h,牛膝总皂苷各组IL-1β、TNF-α水平均明显低于空白组(P均<0.05),且牛膝总皂苷浓度越高,IL-1β、TNF-α水平越低,各组间两两比较差异均有统计学意义(P均<0.05)。结论牛膝总皂苷体外能有效促进人膝骨关节炎软骨细胞增殖,抑制软骨细胞凋亡及软骨细胞中IL-1β、TNF-α的表达。Objective It is to observe the effects of different concentrations of total saponins of Achyranthes bidentata(TSAB)on the proliferation and apoptosis of human knee osteoarthritis chondrocytes and the expression of inflammatory factors interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α).Methods The experiment was divided into a blank group,TSAB 0.01 mg/mL group,TSAB 0.05 mg/mL group,TSAB 0.1 mg/mL group,TSAB 0.5 mg/mL group,and TSAB 1 mg/mL group.The third-generation human knee osteoarthritis chondrocytes were taken,which were routinely cultured in the blank group,and were cultured in each TSAB groups after given corresponding concentration of Achyranthes bidentata saponins D/F12,the cell proliferation and apoptosis were detected by CCK-8 method and Annexin V-FITC/PI,the level of IL-1βwas detected by radioimmunoassay method and the level of TNF-αwas detected by ELISA.Results The cell proliferation of each group of TSAB reached a peak on the 3rd day of culture(the blank group reached the peak on the 5th day),and then showed a downward trend.The OD values of cell proliferation in the TSAB 0.5 mg/mL group were significantly higher than other groups at each time point(all P<0.05).After 3 days of culture,there was no significant difference in the early and late cell apoptosis rate and total apoptosis rate between the 0.01 mg/mL and 0.05 mg/mL TSAB groups and the blank group(all P>0.05);the early and late cell apoptosis rate and total apoptosis rate of TSAB 0.1 mg/mL,0.5 mg/mL,1 mg/mL groups were significantly lower than those of the blank group(all P<0.05),and the total cell apoptosis rate of TSAB 0.5 mg/mL group was significantly lower than those of the 0.1 mg/mL and 1mg/mL groups(all P<0.05).After 72 hours of intervention,the levels of IL-1βand TNF-αin each group of TSAB were significantly lower than those in the blank group(all P<0.05),and the higher the concentration of TSAB,the lower the levels of IL-1βand TNF-α,the differences between the groups were statistically significant(P<0.05).Conclusion TSAB

关 键 词：牛膝总皂苷 骨关节炎 软骨细胞 增殖 凋亡 

分 类 号：R684.3[医药卫生—骨科学]