摘要
目的:观察补肾通络方对骨性关节炎大鼠模型膝关节软骨半胱氨酸天冬氨酸特异性蛋白酶3表达的调节作用。方法:实验于2005-05/2006-03在上海中医药大学动物中心实验室完成。将30只雄性清洁级SD大鼠抽签随机分为正常对照组、骨关节炎模型组、补肾通络方组,每组10只。造模方法:氯胺酮经腹腔注射,麻醉后纵行切开右膝部内侧皮肤,切断内侧副韧带,打开关节腔,切除内侧半月板,切断前后交叉韧带,缝合皮肤。干预方法:正常对照组随意放养,不做处理。补肾通络方药物组成:杜仲、山萸肉、熟地黄、秦艽、寄生、独活等,由上海中医药大学脊柱病研究所提供。自造模第2周开始灌药,2次/d,连续4周。骨性关节病模型组自造模第2周开始灌服等量生理盐水。干预4周后,拉颈处死动物。电镜观察细胞微观形态,免疫组化法检测关节软骨半胱氨酸天冬氨酸特异性蛋白酶3蛋白表达,阳性表达为棕黄色,主要存在于细胞浆内。用平均灰度值和平均吸光度对阳性信号进行量化。RT-PCR法检测半胱氨酸天冬氨酸特异性蛋白酶3mRNA的表达强度。结果:30只大鼠均进入结果分析,无脱失。①半胱氨酸天冬氨酸特异性蛋白酶3表达:骨性关节病模型组明显高于正常对照组,差异非常显著(平均灰度值:142.07±4.04,195.65±2.71,P<0.01;平均吸光度:0.25±0.01,0.12±0.01,P<0.01);补肾通络方组与骨性关节病模型组相比,差异非常显著[平均灰度值:167.62±2.70,142.07±4.04,P<0.01;平均吸光度:0.18±0.01,0.12±0.01,P<0.01]。②半胱氨酸天冬氨酸特异性蛋白酶3mRNA表达:骨性关节病模型组明显高于正常对照组,差异非常显著(3.16±0.26,0.19±0.03,P<0.01);补肾通络方组与骨性关节病模型组相比,差异非常显著(0.41±0.05,3.16±0.26,P<0.01)。结论:补肾通络方能够下调膝骨性关节炎半胱氨酸天冬氨酸特异性蛋白酶3的表达。
AIM： To investigate the expression of caspase-3 in the chondrocyte of osteoarthritis （OA） rats treated by Bushentongluo recipe.
METHODS： The experiment was condt,cted in the Laboratory of Experimental Animal Centre, Shanghai University of Traditional Chinese Medicine from May 2005 to March 2006. Thirty male SD rats of cleaning grade were randomly divided into normal group, OA model group and Bushentongluo recipe group, with 10 rats in each. Modeling： The rats were subjected to anterior cruciate ligament transection （ACLT） of the right knee via microtubule after intraperitoneal injection of ketamine, and medial collateral ligament and medial meniscus were excised, then the skin was sutured. Intervention： The normal control rats were fed freely without management. Bushentongluo recipe： eucommia bark, fructus corni, prepared rehmanniae root, largeleaf gentian root, parasitism and doubleteeth pubescent angelica root, providing by Spondylopathy Institute, Shanghai University of Traditional Chinese Medicine. From the second week of modeling, the administration was given twice a day for continuous 4 weeks, while the OA model group was injected with equal saline. All the rats were executed after four weeks of intervention. Transmission electron microscopy was used for the uhrastrueture, and immunohistochemical and RT-PCR were used to detect the expression of caspase-3 and the level of the caspase-3 mRNA, respectively. The positive expression located in intracytoplasm and colored buffy. And quantitative analysis was conducted by average grey scale value and optical density （OD）.
RESULTS： All the 30 rats were involved in the analysis results without loss.①Expression of caspase-3： The expression was significantly increased in model group, compared ,with control group （average grey scale value： 142.07±4.04, 195.65±2.71, P 〈 0.01; OD： 0.25±0.01, 0.12±0.01, P 〈 0.01）, and was significantly down-regulated by Bushentongluo recipe compared with that of model group （average grey scale value： 167.62±2.70, 142.07±4.04, P 〈 0.01; OD： 0.18±0.01, 0.12±0.01, P 〈 0.01）.②Expression of caspase-3 mRNA was significantly higher in model group than in control group （3.16±0.26, 0.19±0.03, P 〈 0.01）, and was significantly lower in Bushentongluo recipe group than in model group （0.41±0.05, 3.16±0.26, P 〈 0.01）.
CONCLUSION： Bushentongluo recipe can decrease the expression of caspase-3 in OA rats.
出处
《中国临床康复》
CSCD
北大核心
2006年第43期67-69,F0003,共4页
Chinese Journal of Clinical Rehabilitation
基金
上海市重点学科建设项目(T0303)~~
