不同年龄大鼠终板软骨细胞抗氧化能力研究

Study on antioxidant capacity of endplate chondrocytes from different ages in rats

中文关键词:  终板软骨细胞  总抗氧化能力  衰老  凋亡

英文关键词:Endplate chondrocytes  Antioxidant capacity  Ageing  Apoptosis

中文摘要:

  【摘要】 目的：通过研究不同年龄大鼠的终板软骨细胞氧化应激的差异，探讨衰老对终板软骨细胞抗氧化能力的影响。方法：原代培养获取SD大鼠2个月、18个月来源的终板软骨细胞鉴定后，使用第三代细胞进行实验。实验分为两组，A组为2个月大鼠来源的终板软骨细胞，B组为18个月大鼠来源的终板软骨细胞。待A、B两组细胞贴壁后，通过β-半乳糖脱氢酶染色和RT-PCR对端粒酶长度进行检测，比较两组细胞衰老的差异；通过总抗氧化能力试剂盒检测两组细胞总抗氧化能力的不同。以浓度为200umol/L过氧化氢对细胞进行2.5h的处理，RT-PCR法检测处理前后两组细胞Ⅱ型胶原、蛋白聚糖、caspase-3及B淋巴细胞瘤-2（B-cell lymphoma-2，bcl-2）的mRNA表达情况，使用流式细胞仪检测两组细胞的凋亡情况。对Ⅱ型胶原、蛋白聚糖、caspase-3及bcl-2的相对表达量进行组间t检验，P<0.05为差异有统计学意义。结果：未使用过氧化氢处理时，A、B两组终板软骨细胞的在β-半乳糖脱氢酶染色、Ⅱ型胶原、蛋白聚糖表达及凋亡率无明显差异（P>0.05），而B组在端粒酶的相对长度（T/S=10±2）及总抗氧化能力（0.72±0.18）较A组（T/S=135±5，1）显著降低（P<0.05）。在相同条件的过氧化氢刺激后，B组终板软骨细胞的Ⅱ型胶原（0.60±0.16）、蛋白聚糖（0.75±0.22）表达明显低于A组细胞（P<0.05），而凋亡率显著高于A组细胞（P<0.05）。结论：不同年龄大鼠终板软骨细胞衰老及抗氧化能力存在差异，衰老越明显的终板软骨细胞，总抗氧化能力越低，抵抗过氧化损伤的能力越差。

英文摘要:

  【Abstract】 Objectives: To explore the antioxidant capacity of aging on the endplate chondrocytes′ oxidative stress action in rats. Methods: Two kinds of endplate cartilage cells in primary generation were cultured. Cells were tested and passaged to the third generation for experiments. The cells were divided into two groups: group A was endplate chondrocytes from two-month rats; group B was endplate chondrocytes from 18-month rats. β-galactose dehydrogenase staining and PCR test on the relative length of telomere were used to compare their age difference. The total antioxidant capacity of the cell was determined by the total antioxidant capacity kits. Cells were intervented by a concentration of 200mmol/L of hydrogen peroxide for 2.5 hours. Before and after intervention by the same dose of hydrogen peroxide, RT-PCR was used to detect the expression of type Ⅱ collagen, proteoglycans, caspase-3 and bcl-2. Apoptosis of two different cells were compared by flow cytometry. The relative expressions of type Ⅱ collagen, proteoglycans, caspase-3 and bcl-2 were statistically analyzed by t test between two groups, P<0.05 meant statically significant. Results: Before intervention by the same dose of hydrogen peroxide, β-galactose dehydrogenase staining, expressions of type Ⅱ collagen, proteoglycans and apoptosis rate had no significant differences between 2 groups of endplate cartilage cells. The relative length of telomere was found significantly shorten in the aging rat origin and total antioxidant capacity was found significantly lower in group B. After the same dose of hydrogen peroxide stimulation, the expressions of type Ⅱ collagen and proteoglycans in group B were significantly lower than thoses in group A and the level of apoptosis rate was significantly higher than that in group A. Conclusions: Endplate cartilage cell under different ages has different antioxidant capacities. The older of the endplate cartilage cells, the lower ability to resist peroxidation.