阳和汤干预兔骨关节炎软骨细胞中血管内皮生长因子变化

1、阳和汤干预兔骨关节炎软骨细胞中血管内皮生长因子变化         09-08-13 16:05:00     作者：陈朝蔚, 陈永强    编辑：studa20【摘要】  目的：观察阳和汤对兔骨关节炎软骨细胞中血管内皮生长因子（vascular endothelial growth factor, VEGF）表达的影响。方法：新西兰大白兔15只随机分为正常组、模型组和阳和汤组。按照Hulth法建立膝骨关节炎模型，术后第5周起

2、至第7周末，阳和汤组予混有阳和汤的颗粒饲料，正常组及模型组均给予普通饲料；术后第8周分别取各组胫骨平台关节软骨，采用番红O染色，进行Mankin评分；免疫组织化学染色测定关节软骨中VEGF表达的阳性指数。结果：正常对照组番红O染色的Mankin评分与模型组比较，差异有统计学意义（P<0.01）。阳和汤组番红O染色减少或缺失程度较模型组轻，差异有统计学意义（P<0.01）。各组关节软骨切片VEGF免疫组化染色分析，模型组VEGF的阳性指数明显高于正常组，差异有统计学意义（P<0.01），阳和汤组VEGF阳性指数明显低于模型组，差异有统计学意义（P<0.05）。结论：VEG

3、F在骨关节炎形成早期起重要作用；阳和汤可延缓关节软骨退行性改变，其作用可能是通过调控VEGF而达到抑制血管增生的作用。 【关键词】  骨关节炎; 阳和汤; 软骨; 血管内皮生长因子; 血管生成, 病理性Objective: To observe the effects of Yanghe Decoction on vascular endothelial growth factor (VEGF) in cartilage cells of osteoarthritis rabbits.Methods: Fifteen New Zealand white rabbits were ra

4、ndomly divided into normal group, untreated group and Yanghe Decoctiontreated group. The rabbit model of osteoarthritis was established according to Hulth's method. The rabbits were sacrificed at the 8th week after administration of Yanghe Decoction for 14 days, and then rabbit tibia articular c

5、artilage was removed. Sections of the cartilage were stained with Safranin O for histological examination. The cartilage histological characteristics were observed according to the method of Mankin. Immunohistochemical staining was performed to investigate the expression of VEGF. Articular cartilage

6、s were observed with microscopy and image analysis method was used to measure the expression intensity of VEGF.Results: There were significant differences in Mankin score between normal group and untreated group (P<0.01), and between untreated group and Yanghe Decoctiontreated group (P<0.01).

7、Immunohistochemical staining indicated that the expression intensity of VEGF in untreated group was significantly increased as compared with that in normal group (P<0.01), and also obviously higher than Yanghe Decotiontreated group.Conclusion: VEGF plays an important role during early stage of OA

8、. Yanghe Decoction can protect the articular cartilage through suppressing the VEGF expression in chondrocytes and then suppress angiogenesis.Keywords: osteoarthritis; Yanghe Decoction; cartilage; vascular endothelial growth factor; angiogenesis, pathologic    骨性关节炎（osteoarthritis, OA

9、）的临床表现以关节疼痛、僵硬和功能障碍为主要症状。其基本病理变化为关节软骨和软骨下骨损伤、骨质增生及关节内组织的炎性病变。对于骨性关节炎病因现在尚未明确。目前国内外研究已证实骨性关节炎和血管发生的关系密切，骨性关节炎受累滑膜和软骨中已发现多种血管生成介质1，其中血管内皮生长因子（vascular endothelial growth factor, VEGF）是新生血管形成的重要调节因子。阳和汤在临床使用中疗效较好，本文旨在探讨阳和汤对兔膝骨关节炎软骨细胞中VEGF的影响及其作用机制，为临床应用提供部分实验依据。1  材料和方法1.1  动物及试剂 6月龄新西兰大

10、白兔15只，雌雄兼用，体质量（2.3±0.4）kg，由上海中医药大学实验动物中心提供，动物合格证号为SCXK（沪）20040004，清洁级。随机分为正常组、模型组、阳和汤组，每组5只。阳和汤功用温阳补血，散寒通滞，出自清代王洪绪外科证治全生集。药物组成：熟地黄30 g、肉桂3 g、麻黄2 g、鹿角胶9 g、白芥子6 g、姜炭2 g、生甘草3 g，由上海市中医医院制剂科提供。VEGF第一抗体购于美国Santa Cruz公司。1.2  OA模型建立 模型组与阳和汤组按照Hulth等2报道的方法建立膝骨关节炎模型：3%戊巴比妥钠（30 mg/kg）耳缘静脉麻醉，麻醉满意后，手术切开兔左膝关节，剪断内侧副韧带和前后交叉韧带，完整切除内侧半月板，术中注意保护关节软骨面不受损伤；彻底止血，逐层缝合关节囊、皮下和皮肤等组织，不予固定。正常组不予任何手术处理。1.3  给药方式及剂量 从术后第5周起至第7周末结束。阳和汤组：阳和汤方水煎浓缩后与兔饲料混合均匀，制成定量颗粒饲料，予40 g/d相当于生药6 g/（kg·d），按体表面积折算。正常组和模型组予40 g/d普通