| 鹿茸多肽对抗骨关节炎软骨细胞氧化损伤作用的实验研究 |
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投稿时间：2010-08-24
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| 期刊信息：《中国骨伤》2011年，第24卷，第3期，第245-248页 |
| DOI：10.3969/j.issn.1003-0034.2011.03.020 |
| 基金项目:国家自然科学基金资助项目(编号：30472224) |
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中文摘要:
目的：研究鹿茸多肽对骨关节炎软骨细胞的氧化损伤的逆转作用，探讨鹿茸多肽保护软骨细胞的主要作用机制。
方法：选15只5月龄日本大耳白兔，采用内侧副韧带、前后交叉韧带切断并切除内侧半月板的Hulth骨性关节炎动物模型制作方法，体外分离培养软骨细胞。以假手术组细胞为正常对照组细胞，造模组细胞为骨关节炎软骨细胞，分别加入6.25、12.5、25 μg/ml鹿茸多肽低、中、高剂量，从第9周(2个月)开始，每周处死1组动物，分离培养软骨细胞。连续8周，DCFH-DA法检测细胞内活性氧水平，Griess法测定细胞培养上清中NO、SOD和GSH-Px含量。
结果：DCFH-DA法检测细胞内活性氧水平，对照组平均5.46±0.46,模型组平均12.08±0.74,两组比较，P<0.001;鹿茸多肽低、中、高剂量组分别为(9.81±0.59)、(7.83±0.63)和(6.89±0.71),与模型组比较，差异均有统计学意义(P<0.05).骨关节炎模型组中NaNO2、SOD、GSH-Px分别为(5.60±0.45) μM、(38.56±12.53) U/ml和(151.90±25.60) U,与对照组比较，差异均有统计学意义(P<0.001,P<0.05).NaNO2水平鹿茸多肽低剂量组(4.34±0.39) μM,中剂量组(3.67±0.36) μM,高剂量组(3.20±0.27) μM,与模型组比较差异均有统计学意义(P<0.01);SOD水平鹿茸多肽低剂量组(49.91±5.77)U/ml,中剂量组(54.05±5.27)U/ml,高剂量组(57.44±5.70) U/ml,与模型组比较差异均有统计学意义(P<0.05);GSH-Px水平鹿茸多肽低剂量组(172.50±18.65) U,中剂量组(202.10±21.60) U,高剂量组(315.80±10.50) U,中剂量组及高剂量组与模型组比较差异均有统计学意义(P<0.01).
结论：鹿茸多肽对骨关节炎软骨细胞的氧化损伤有逆转作用，且在一定范围内呈现剂量依赖性。鹿茸多肽这种抗氧化损伤作用，具有研发成为骨性关节炎治疗药物的潜能。 |
| 【关键词】鹿茸多肽 骨性关节炎 软骨细胞 氧化损伤 动物实验 |
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| Experimental study of velvet antler polypeptides against oxidative damage of osteoarthritis cartilage cells |
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ABSTRACT
Objective: To study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides(VAPS),and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.
Methods: Fifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut,articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group,osteoarthritis cartilage cells in the model groups were added VAPS 6.25,12.5,25 μg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA,the content of NO,SOD and GSH-Px in cell culture supernatant were detected by Griess method.
Results: DCFH-DA detection of intracellular reactive oxygen species was(5.46±0.46)in the control group,(12.08±0.74) in the model groups. The model group compared with the control group by t test with the P value less than <0.001. DCFH-DA detection of intracellular reactive oxygen species was(9.81±0.59)in VAPS 6.25 μg/ml group,(7.83±0.63)in the VAPS 12.5 μg/ml group,(6.89±0.71)in the VAPS 25 μg/ml group,as compared with model group there were statistically significant difference(P<0.05). The content of NaNO2,SOD and GSH-Px in osteoarthritis model group was(5.60±0.45) μM,(38.56±12.53) U/ml and (151.90±25.60) U,as compared with control group there were statistically significant difference(P<0.001,P<0.05);The content of NaNO2 was(4.34±0.39) μM in VAPS 6.25 μg/ml group,(3.67±0.36) μM in the VAPS 12.5 μg/ml group,(3.20±0.27) μM in the VAPS 25 μg/ml group,as compared with model group there were statistically significant difference(P<0.01). The content of SOD was(49.91±5.77) U/ml in VAPS 6.25 μg/ml group,(54.05±5.27) U/ml in the VAPS 12.5 μg/ml group,(57.44±5.70) U/ml in the VAPS 25 μg/mL group,as compared with model group there was statistically significant (P<0.05). The content of GSH-Px was (172.50±18.65) U in VAPS 6.25 μg/ml group,(202.10±21.60) U in the VAPS 12.5 μg/ml group,(315.80±10.50) U in the VAPS 25 μg/ml group,the VAPS 12.5 μg/mL group and VAPS 25 μg/ml group was compared with model group,there were statistically significant difference(P<0.01).
Conclusion: The VAPS have anti-oxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis. |
| KEY WORDS Velvet antler polypeptides Osteoarthritis Chondrocytes Oxidative damage Animal experimentation |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 李振华,赵文海,周秋丽.鹿茸多肽对抗骨关节炎软骨细胞氧化损伤作用的实验研究[J].中国骨伤,2011,24(3):245~248 |
| 英文格式: | LI Zhen-hua,ZHAO Wen-hai,ZHOU Qiu-li.Experimental study of velvet antler polypeptides against oxidative damage of osteoarthritis cartilage cells[J].zhongguo gu shang / China J Orthop Trauma ,2011,24(3):245~248 |
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