CAJ | 학술논문

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、0.0625、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dUTP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（ P＜0.05）。 SNP浓度为0.0625、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 （ P＜0.05）； 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（ P＜0.05）。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（ P＜0.05）。 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 （ P＜0.05）；且不同浓度间两两比较，差异有统计学意义（P＜0.05）。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。
목적：비교불동농도초보납（ SNP）유도토연골세포조망모형，위진일보탐토약물대연골세포조망적조공작용제공이론지지。방법2014년9—12월선취4주령건강웅성신서란대이백토8지，체질량200～250 g，체외분리배양병감정토연골세포，채용불동농도SNP （0、0.0625、0.125、0.25、0.5、1、2 mmol／L）유도토연골세포，분별우12、24、48 h채용사갑기우담서염비색시험법（ MTT법）화말단탈양핵감산전이적dUTP결구말단표기법（ TUNEL법）평개병비교불동농도SNP유도연골세포조망모형，기록토연골세포증식정황（ OD치）화토연골세포조망솔。결과불동농도SNP유도적토연골세포재불동시간점적 OD치비교，차이균유통계학의의（ P＜0.05）。 SNP농도위0.0625、0.125 mmol／L배양12、24、48 h시토연골세포OD치고우SNP농도위0 mmol／L （ P＜0.05）； SNP농도위0.25、0.5、1、2 mmol／L배양12、24、48 h시토연골세포OD치균저우SNP농도위0 mmol／L； SNP농도위0.25、0.5、1、2 mmol／L배양12、24、48 h시토연골세포OD치재불동농도간량량비교，차이균유통계학의의（ P＜0.05）。불동농도SNP유도적토연골세포재불동시간점적조망솔비교，차이균유통계학의의（ P＜0.05）。 SNP농도위0.25、0.5、1、2 mmol／L배양12、24、48 h시토연골세포조망솔균고우SNP농도위0 mmol／L （ P＜0.05）；차불동농도간량량비교，차이유통계학의의（P＜0.05）。결론 SNP농도위0.25～2 mmol／L차유도12、24、48 h시가제비불동엄중정도적연골세포조망모형，위진일보탐토약물대연골세포조망적조공작용제공료이론지지。
Objective To compare the chondrocytes apoptosis models induced by different concentrations of sodium nitroprusside ( SNP) , in order to provide theoretical support for the further investigation of the regulating effect of drugs on chondrocytes apoptosis.Methods Eight 4-week-old healthy New Zealand male rabbits (weight, 200-250 g) were selected from September to December in 2014.Rabbit chondrocytes were cultured and researched in vitro .Different concentrations of SNP (0.062 5, 0.125, 0.25, 0.5, 1, 2 mmol/L) were used to induce rabbit chondrocytes .At 12, 24 and 48 h, MTT method and TUNEL method were employed to evaluate the chondrocytes apoptosis models induced by different concentrations of SNP .The OD value and apoptosis rate of rabbit chondrocytes were recorded .Results At different time points , chondrocytes apoptosis models induced by different concentrations of SNP were significantly different ( P<0.05) in OD value.At 12, 24 and 48 h, chondrocytes apoptosis models induced by 0.062 5 mmol/L and 0.125 mmol/L of SNP were higher ( P <0.05 ) than the chondrocytes apoptosis model induced by 0 mmol/L of SNP in OD value; at 12, 24 and 48 h, chondrocytes apoptosis models induced by 0.25, 0.5, 1 and 2 mmol/L of SNP were lower ( P<0.05) than the chondrocytes apoptosis model induced by 0 mmol/L of SNP in OD value; at 12, 24 and 48 h, chondrocytes apoptosis models induced by 0.25, 0.5, 1 and 2 mmol/L of SNP were significantly different ( P<0.05 ) in OD value.At different time points , chondrocytes apoptosis models induced by different concentrations of SNP were significantly different ( P <0.05) in apoptosis rate.At 12, 24 and 48 h, chondrocytes apoptosis models induced by 0.25, 0.5, 1 and 2 mmol/L of SNP were higher (P<0.05) than the chondrocytes apoptosis model induced by 0 mmol/L of SNP in apoptosis rate; chondrocytes apoptosis models induced by different concentrations of SNP were significantly different ( P<0.05) in apoptosis rate.Conclusion Chondrocytes apoptosis models of different severity degrees can be build by SNP with the concentration ranging from 0.25 mmol/L to 2 mmol/L, which provides theoretical support for the regulating effect of drugs on chondrocytes apoptosis .