不同浓度硝普钠诱导兔软骨细胞凋亡模型的比较研究

发布时间：2018-10-30 15:17

【摘要】：目的比较不同浓度硝普钠(SNP)诱导兔软骨细胞凋亡模型,为进一步探讨药物对软骨细胞凋亡的调控作用提供理论支持。方法 2014年9—12月选取4周龄健康雄性新西兰大耳白兔8只,体质量200~250 g,体外分离培养并鉴定兔软骨细胞,采用不同浓度SNP(0、0.062 5、0.125、0.25、0.5、1、2 mmol/L)诱导兔软骨细胞,分别于12、24、48 h采用四甲基偶氮唑盐比色试验法(MTT法)和末端脱氧核苷酸转移的d UTP缺口末端标记法(TUNEL法)评价并比较不同浓度SNP诱导软骨细胞凋亡模型,记录兔软骨细胞增殖情况(OD值)和兔软骨细胞凋亡率。结果不同浓度SNP诱导的兔软骨细胞在不同时间点的OD值比较,差异均有统计学意义(P0.05)。SNP浓度为0.062 5、0.125 mmol/L培养12、24、48 h时兔软骨细胞OD值高于SNP浓度为0 mmol/L(P0.05);SNP浓度为0.25、0.5、1、2 mmol/L培养12、24、48 h时兔软骨细胞OD值均低于SNP浓度为0 mmol/L;SNP浓度为0.25、0.5、1、2 mmol/L培养12、24、48 h时兔软骨细胞OD值在不同浓度间两两比较,差异均有统计学意义(P0.05)。不同浓度SNP诱导的兔软骨细胞在不同时间点的凋亡率比较,差异均有统计学意义(P0.05)。SNP浓度为0.25、0.5、1、2 mmol/L培养12、24、48 h时兔软骨细胞凋亡率均高于SNP浓度为0 mmol/L(P0.05);且不同浓度间两两比较,差异有统计学意义(P0.05)。结论 SNP浓度为0.25~2 mmol/L且诱导12、24、48 h时可制备不同严重程度的软骨细胞凋亡模型,为进一步探讨药物对软骨细胞凋亡的调控作用提供了理论支持。
[Abstract]:Objective to compare the model of rabbit chondrocyte apoptosis induced by sodium nitroprusside (SNP) at different concentrations, and to provide theoretical support for the further study of the regulation of chondrocyte apoptosis by drugs. Methods from September to December 2014, 8 healthy male New Zealand white rabbits of 4 weeks of age, weighing 200,250g, were isolated and identified as chondrocytes in vitro. The chondrocytes were isolated and identified with different concentrations of SNP. Mmol/L) induced rabbit chondrocytes, The apoptotic model of chondrocytes induced by different concentrations of SNP was evaluated and compared with tetramethylazolium colorimetric assay (MTT) and d UTP Nick end labeling (TUNEL) for terminal deoxynucleotide transfer. The proliferation of rabbit chondrocytes (OD) and the rate of chondrocyte apoptosis were recorded. Results the OD values of rabbit chondrocytes induced by different concentrations of SNP were compared at different time points. The difference was statistically significant (P0.05). SNP concentration was 0.062, 0.125 mmol/L culture for 48 h, the OD value of rabbit chondrocytes was higher than that of SNP concentration was 0 mmol/L (P0.05); The OD values of chondrocytes in rabbit chondrocytes were lower than that of SNP concentration 0 mmol/L; after cultured for 48 h with SNP concentration of 0.25 and 0.5 mmol/L. The OD value of rabbit chondrocytes was significantly higher than that of rabbit chondrocytes at different concentrations (P0.05) when the concentration of SNP was 0.25 and 0.5 / 1 / 2 mmol/L cultured for 48 h (P < 0.05) and the OD values of chondrocytes in rabbits were significantly different (P0.05). The apoptosis rate of rabbit chondrocytes induced by SNP at different concentration was significantly different at different time points (P0.05). SNP concentration was 0.25). SNP 0.51, P 0.05), and the apoptosis rate of rabbit chondrocytes induced by SNP at different time points was significantly different (P0.05). 2 the apoptotic rate of rabbit chondrocytes was higher than that of SNP (0 mmol/L) at 48 h after mmol/L culture (P0.05). And different concentrations of pairwise comparison, the difference was statistically significant (P0.05). Conclusion the model of chondrocyte apoptosis with different severity can be established when the concentration of SNP is 0.25 ~ 2 mmol/L and induced for 48 h, which provides theoretical support for further study on the regulation of chondrocyte apoptosis by drugs.
【作者单位】：辽宁医学院药学院;辽宁医学院附属第一医院;中国科学院上海药物研究所;
【基金】：国家自然科学基金资助项目(81302681) 国家“十二五”重大新药创制基金(2012ZX093016-002) 辽宁省自然科学基金资助项目(2013022051) 辽宁医学院校长基金-奥鸿博泽基金(XZJJ20130106-02);辽宁医学院校长基金-全民口腔研究生科研创新基金(QM2014007)
【分类号】：R965

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