兔关节软骨细胞的培养及硝普钠对其凋亡影响的实验研究
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摘要
目的:熟练掌握实验室相关仪器的操作,培养无菌操作的实验观念,进行兔关节软骨细胞的分离及培养,观察兔关节软骨细胞体外培养的生物学形态,进行不同浓度的硝普钠作用不同的时间后诱导兔关节软骨细胞凋亡的实验,并检测其凋亡率,来探讨一氧化氮(NO)供体硝普钠(SNP)诱导关节软骨细胞凋亡的机制及关节软骨细胞凋亡与骨关节炎(OA)的发生机制。
方法:
1.在实验室进行不同实验仪器的操作。
2.在实验室进行无菌操作的动手锻炼。
3.在净化操作台上进行各种实验试剂的配制。
4.在净化操作台上进行兔关节软骨细胞的分离和培养。
5.在CO_2培养箱内进行兔关节软骨细胞的培养。
6.在显微镜下观察兔关节软骨细胞的生物学形态。
7.在净化操作台上进行不同浓度的硝普钠作用不同的时间后诱导兔关节软骨细胞凋亡的实验操作。
8.用流氏细胞仪检测兔关节软骨细胞凋亡,并对实验结果进行统计学分析。
结果:
1.对兔关节软骨细胞生物学形态有了初步的认识。
2.成功建立NO供体SNP诱导软骨细胞凋亡的模型
3.软骨细胞凋亡与SNP有一定的量效和时效关系。
4.不同的浓度组,不同的时间组采用组间配伍组设计资料的方差分析具有统计学意义(p＜0.01)。
5.2mmol/L浓度的硝普钠诱导兔关节软骨细胞凋亡,培养24小时后,其凋亡率明显高于其他浓度和其他时间的对照组(p＜0.01)。
结论:通过成功建立NO供体SNP诱导软骨细胞凋亡的模型,发现软骨细胞凋亡与SNP有一定的量效和时效关系,有助于了解NO引起关节软骨细胞凋亡的发生机制及关节软骨细胞凋亡与OA的发生机制。
OBJECTIVE:To master the operation of related equipment,to train the experimental concept of asepsis,to carry out the separation and culture of articular chondrocytes from rabbit,to observe the biological patter of articular chondrocytes from rabbit in vitro, to carry out the experiment of of articular chondrocytes from rabbit induced by different concentrations of sodium nitroprusside(SNP) and different times,to calculate of the rate of apoptosis,to explore the mechanisms of apoptosis of articular chondrocytes induced by sodium nitroprusside(SNP) and the mechanisms of occurrence between apoptosis of articular cartilage and osteoarthritis(OA).
METHODS:
1.Operate the different devices in the laboratory..
2.Carry out the training of aseptic operation in the laboratory.
3.Make up a variety of experimental reagents in clean console.
4.Carry out the separation and culture of articular chondrocytes from rabbit in CO_2 incubator.
5.Carry out the culture of articular chondrocytes from rabbit in the CO_2 incubator.
6.Observe the biological morphology of articular chondrocytes from rabbit under the microscope.
7.Carry out the experiment of of articular chondrocytes from rabbit induced by different concentrations of sodium nitroprusside(SNP) and different times.
8.Detect the apoptosis of articular chondrocytes from rabbit by low cytometry.
RESULTS:
1.On biology of articular chondrocytes from rabbits with an initial understanding of the form.
2.Set up the cartilage model of apoptosis successfully ihduced by NO donor SNP.
3.Chondrocytes apoptosis and the SNP have a relationship between concentration and time.
4.Use the inter-group design information combined analysis of variance is significant in statistics between the groups of different concentration and the groups of different times(p <0.01).
5.Apoptosis of articular chondrocytes from rabbit induced by 2mmol /L concentration of sodium nitroprusside and cultured for 24 hours, the rate of apoptosis is significantly higher than other concentrations and other times of the control group(p <0.01).
CONCLUSION:Establish the model of NO donor SNP-induced chondrocyte apoptosis sccessfully and found that SNP has a certain amount of efficiency and timeliness,to understand NO cause the apoptosis of articular chondrocytes and the mechanism of apoptosis of articular chondrocytes and OA.
方法:
1.在实验室进行不同实验仪器的操作。
2.在实验室进行无菌操作的动手锻炼。
3.在净化操作台上进行各种实验试剂的配制。
4.在净化操作台上进行兔关节软骨细胞的分离和培养。
5.在CO_2培养箱内进行兔关节软骨细胞的培养。
6.在显微镜下观察兔关节软骨细胞的生物学形态。
7.在净化操作台上进行不同浓度的硝普钠作用不同的时间后诱导兔关节软骨细胞凋亡的实验操作。
8.用流氏细胞仪检测兔关节软骨细胞凋亡,并对实验结果进行统计学分析。
结果:
1.对兔关节软骨细胞生物学形态有了初步的认识。
2.成功建立NO供体SNP诱导软骨细胞凋亡的模型
3.软骨细胞凋亡与SNP有一定的量效和时效关系。
4.不同的浓度组,不同的时间组采用组间配伍组设计资料的方差分析具有统计学意义(p＜0.01)。
5.2mmol/L浓度的硝普钠诱导兔关节软骨细胞凋亡,培养24小时后,其凋亡率明显高于其他浓度和其他时间的对照组(p＜0.01)。
结论:通过成功建立NO供体SNP诱导软骨细胞凋亡的模型,发现软骨细胞凋亡与SNP有一定的量效和时效关系,有助于了解NO引起关节软骨细胞凋亡的发生机制及关节软骨细胞凋亡与OA的发生机制。
OBJECTIVE:To master the operation of related equipment,to train the experimental concept of asepsis,to carry out the separation and culture of articular chondrocytes from rabbit,to observe the biological patter of articular chondrocytes from rabbit in vitro, to carry out the experiment of of articular chondrocytes from rabbit induced by different concentrations of sodium nitroprusside(SNP) and different times,to calculate of the rate of apoptosis,to explore the mechanisms of apoptosis of articular chondrocytes induced by sodium nitroprusside(SNP) and the mechanisms of occurrence between apoptosis of articular cartilage and osteoarthritis(OA).
METHODS:
1.Operate the different devices in the laboratory..
2.Carry out the training of aseptic operation in the laboratory.
3.Make up a variety of experimental reagents in clean console.
4.Carry out the separation and culture of articular chondrocytes from rabbit in CO_2 incubator.
5.Carry out the culture of articular chondrocytes from rabbit in the CO_2 incubator.
6.Observe the biological morphology of articular chondrocytes from rabbit under the microscope.
7.Carry out the experiment of of articular chondrocytes from rabbit induced by different concentrations of sodium nitroprusside(SNP) and different times.
8.Detect the apoptosis of articular chondrocytes from rabbit by low cytometry.
RESULTS:
1.On biology of articular chondrocytes from rabbits with an initial understanding of the form.
2.Set up the cartilage model of apoptosis successfully ihduced by NO donor SNP.
3.Chondrocytes apoptosis and the SNP have a relationship between concentration and time.
4.Use the inter-group design information combined analysis of variance is significant in statistics between the groups of different concentration and the groups of different times(p <0.01).
5.Apoptosis of articular chondrocytes from rabbit induced by 2mmol /L concentration of sodium nitroprusside and cultured for 24 hours, the rate of apoptosis is significantly higher than other concentrations and other times of the control group(p <0.01).
CONCLUSION:Establish the model of NO donor SNP-induced chondrocyte apoptosis sccessfully and found that SNP has a certain amount of efficiency and timeliness,to understand NO cause the apoptosis of articular chondrocytes and the mechanism of apoptosis of articular chondrocytes and OA.
引文
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