摘要
目的:观察烟酰胺对肿瘤坏死因子α诱导纤维环基质降解的抑制作用,并分析其作用途径。方法:实验于2005-10/12在华中科技大学同济医学院附属协和医院中心实验室及骨科实验室完成。椎间盘组织取自12只清洁级日本大白兔,兔龄4个月,体质量2.5～3.0kg,由华中科技大学同济医学院实验动物中心提供。建立兔椎间盘组织凝胶培养模型,将其分为4组,即正常对照组、烟酰胺组、退变组及治疗组。正常对照组不加入药物;烟酰胺组加入0.5g/L烟酰胺;退变组加入10 μg/L肿瘤坏死因子α诱导纤维环基质降解;治疗组加入0.5g/L烟酰胺和10 μg/L肿瘤坏死因子α。培养1周后比较各组标本纤维环结构(藏红O-快绿染色强度)、糖醛酸含量、Ⅰ,Ⅱ型胶原结构及半胱氨酸天冬氨酸蛋白酶3表达情况(染色阳性细胞率)。结果:①各组兔椎间盘标本纤维环结构比较:治疗组纤维环结构的破坏得到较好的抑制,染色浓度高于退变组。②各组兔椎间盘标本纤维环糖醛酸含量比较:治疗组较退变组增高约48%(t=16.93,P=0.000),与烟酰胺组接近(t=0.71,P=0.657)。③各组兔椎间盘标本纤维环Ⅰ,Ⅱ型胶原结构及分布情况:治疗组板层结构完整性和胶原纹路连续性好于退变组,Ⅰ,Ⅱ型胶原含量之比较正常组升高。④各组兔椎间盘标本纤维环半胱氨酸天冬氨酸蛋白酶3阳性染色率比较:治疗组明显低于退变组(10.3%,17.9%,χ2=5.081,P<0.01)。结论:烟酰胺能够减轻肿瘤坏死因子α对纤维环基质的破坏作用,这种作用与烟酰胺减少纤维环半胱氨酸天冬氨酸蛋白酶3依赖的细胞凋亡有关。
AIM： To investigate the protective effect and mechanism of protection of niacinamide against annulus fibrosus （AF） degeneration that is induced by tumor necrosls factor-α （TNF-α）.
METHODS： The experiment was carried out in the Central Laboratory and Orthopaedic Laboratory, Union Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology between October and December in 2005. Chiba＇s intervertebral disc （IVD） culture models were enrolled from 12 Japanese white rabbits of 4 months old and 2.5-3.0 kg weight, which were offered by the Experimental Animal Center of Tongji Medical College, Huazhong University of Science and Technology. They were randomly divided into 4 groups： control group, niacinamide group （containing 0.5 g/L niacinamide）, degeneration group （10 μg/L TNF-α）, and treatment group （both niacinamide and TNF-α）. After 1-week of culture, AFs were collected for Safranine O staining, Glycosaminoglycan （GS） content measuring, type Ⅰ and Ⅱ collagen as well as cysteinyl aspartate specific protease-3 （Caspase-3） expression （immunohistochemicel examination）.
RESULTS： ①AF structure of rabbit IVD： The destruction of treatment group was inhibited greatly, and the staining was better than that of degeneration group. ②Comparad with degeneration group, the uronic acid content of the treatment group was increased by 48% （t =16.93, P =0.000）, and was similar with that of nlacinamide group （t =0.71, P =0.657）③ The lamellar structure and continuity of Type Ⅰ and Ⅱ collagen was better in treatment group than in degeneration group, and the collagen content was increased compared with control group. ④Rate of Caspase-3 positive staining AF ceils was significantly lower in treatment group than in degeneration group （10.3%, 17.9%, Χ^2=5.081, P 〈 0.01）.
CONCLUSION： Niecinamide can effectively inhibit the apoptosis of Caspase-3 in AFs so as to reduce destruction caused by TNF-α
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第2期305-308,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
作者简介
徐润冰，男．1981年生，湖北省英山县人．汉族，华中科技大学同济医学院在读硕士。主要从事脊柱外科方面的研究。xurunbing@163.com
