桃红四物汤对激素性股骨头缺血坏死模型兔血管内皮生长因子表达和血液流变学的影响

Effects of Taohong Siwu decoction on vascular endothelial growth factor and hemorheology in rabbits with glucocorticoid-induced avascular necrosis of femoral head

目的:观察桃红四物汤对激素性股骨头缺血坏死模型兔血管内皮生长因子的表达和血液流变学的影响,进一步阐明活血化瘀法防治激素性股骨头缺血坏死的作用机制。方法:实验于2005-09/2006-01在福建中医学院中心实验室完成。选择健康成年新西兰大白兔20只,按随机数字表法分为4组,正常对照组、模型对照组、生理盐水对照组、治疗组,每组5只。采用激素注射造模,模型对照组、生理盐水对照组、治疗组兔臀肌注射醋酸氢化泼尼松7.5mg/(kg·只),2次/周;正常对照组兔注射等量生理盐水。所有动物肌注青霉素16万U和链霉素15万U预防感染,2次/周。治疗组给予桃红四物汤(出自清·吴谦等所著的《医宗金鉴》,原方组成为桃仁、红花、当归、生地黄、赤芍、川芎)7mL/kg灌胃,1次/d;生理盐水对照组给予等量生理盐水灌胃。6周后正常对照组和模型对照组麻醉后各处死5只,13周后生理盐水对照组、治疗组麻醉后各处死5只,分别进行股骨头组织病理学观察及血管内皮生长因子表达和血液流变学指标的测定。结果:纳入大白兔20只,均进入结果分析。①股骨头组织病理学观察:模型对照组空缺骨陷窝数高于正常对照组,差异有非常显著性意义[分别为(27.50±5.40)%,(14.50±1.72)%,P<0.001]。治疗组兔股骨头软骨、骨小梁、骨细胞及髓腔组织等均接近正常,空缺骨陷窝数基本恢复到正常对照组水平(P>0.05),治疗组空缺骨陷窝数低于模型对照组,差异有非常显著性意义[分别为(15.80±2.83)%,(27.50±5.40)%,P<0.001];生理盐水对照组与模型对照组病理情形相似,其空缺骨陷窝数明显高于正常对照组和治疗组,差异有非常显著性意义[分别为(28.70±7.26)%,(14.50±1.72)%,(15.80±2.83)%,P<0.001]。②血管内皮生长因子的表达:模型对照组血管内皮生长因子灰度指数(A)低于正常对照组,即表达增强,差异有显著性意义(分别为90.39±4.31,97.58±0.56,P<0.01);治疗组与正常对照组比较差异无显著性意义(P>0.05);生理盐水对照组血管内皮生长因子灰度指数(A)高于正常对照组,表达减弱,差异有非常显著性意义(分别为133.53±4.96,97.58±0.56,P<0.001);治疗组血管内皮生长因子灰度指数(A)低于生理盐水对照组,表达增强,差异有非常显著性意义(分别为97.55±0.74,133.53±4.96,P<0.001);生理盐水对照组血管内皮生长因子灰度指数(A)高于模型对照组,表达减弱,差异有非常显著性意义(分别为133.53±4.96,90.39±4.31,P<0.001)。③血液流变学指标:模型对照组、治疗组和生理盐水对照组全血高切黏度、全血低切黏度、血浆黏度高于正常对照组,差异有显著性意义[分别为(3.35±0.05),(3.14±0.02),(3.50±0.07),(2.76±0.22)mPa·s;(7.11±0.40),(5.77±0.24),(7.48±0.17),(5.49±0.48)mPa·s;(1.70±0.05),(1.57±0.06),(1.72±0.03),(1.48±0.03)mPa·s,P<0.001,0.01,0.05];治疗组全血高切黏度、全血低切黏度、血浆黏度低于模型对照组和生理盐水对照组,差异有非常显著性意义(P<0.001,0.01)。各组间红细胞压积比较,差异均无显著性意义(P>0.05)。生理盐水对照组红细胞聚集指数高于正常对照组,差异有显著性意义(分别为0.54±0.04,0.46±0.04,P<0.01);治疗组红细胞聚集指数低于模型对照组和生理盐水对照组,差异有显著性意义(分别为0.43±0.04,0.50±0.05,0.54±0.04,P<0.05,0.01)。结论:血液流变学异常是激素性股骨头缺血坏死的重要原因之一,活血化瘀中药可对抗激素的作用,间接地促进血管内皮生长因子的表达,可以有效地改善股骨头微循环障碍。

AIM： To investigate the effect on vascular endothelial growth factor （VEGF） and hemorheology of femoral head avascular necrosis induced by glucocorticoid in rabbits by treatment of Taohong Siwu decoction and further explore the mechanism of anti-osteonecrosis by Chinese herbs of activating blood circulation to remove stasis. METHODS： The experiment was carried out in the Central Laboratory of Fujian College of Traditional Chinese Medicine from September 2005 to January 2006. A total of 20 healthy adult rabbits of New Zealand species were randomly divided into 4 groups： normal control group, model control group, saline control group and treatment group, each containing 5 rabbits. Hormone injection was used for model establishment, and each rabbit in normal control group was injected with 7.5 mg/kg saline while the rabbits in other three groups were administered with equal volume of hydroxyprednisone through gluteus injection （twice a week） for 6 weeks. All the animals were injected intramuscularly with 1.6×10^6 U penicillin and 1.5×10^6 U streptomycin for the prevention of infection, twice a week. Then the rabbits in treatment group were given 7 mL/kg Taohong Siwu decoction （peach seed, safflower, angelica, rehmannia, red peony root and Szechwan lovage rhizome, cited from the Golden Mirror of Medicine by Wu Qian in Qing Dysnasty）, once daily; the rabbits in saline control group were given equal volume of saline intragastrically. Six weeks later, 5 rabbits of normal control group and model control group were executed respectively and another seven weeks later 5 rabbits of saline control group and treatment group were executed separately for the measurements of histopathology, VEGF expression and hemorheology of femoral head. RESULTS： Totally 20 rabbits were involved in the result analysis.① Histopathologic measurements showed that empty lacuna increased in model control group and saline control group compared with normal control group and treatment group, with the extremely significant differences [（27.50±5.40）%, （28.70±7.26）%, （14.50±1.72）%, （15.80±2.83）%, P 〈 0.001]. And cartilage, trabecula of bone, osteoeytes and the medullary cavity lipocytes in treatment group showed similar size and amount to those in normal control group （P 〉 0.05）.②The expression of VEGF by grey index （A） increased significantly in model control group compared with normal control group （90.39±4.31, 97.58±0.56, P 〈 0.01）; There was insignificant difference between treatment group and normal control group （P 〉 0.05）; In comparison with saline control group, the expressions in normal control group and treatment group were extremely significantly increased （133.53±4.96, 97.58±0.56, P 〈 0.001; 97.55±0.74, 133.53±4.96,P 〈 0.001）; Additionally, the saline control group presented lower expression of VEGF than model control group, with extremely significant differences （133.53±4.96, 90.39±4.31, P 〈 0.001）.③Hemorheology： Whole blood viscosity and plasma viscosity were significantly higher in model control group, treatment group and saline group than in normal control group [（3.35±0.05）, （3.14±0.02）, （3.50±0.07）, （2.76±0.22） mPa· s; （7.11±0.40）, （5.77±0.24）, （7.48±0.17）, （5.49±0.48） mPa·s; （1.70±0.05）, （1.57±0.06）, （1.72±0.03）, （1.48±0.03） mPa·s, P 〈 0.001, 0.01, 0.05], and extremely significantly lower in treatment group than in model control group and saline group （P 〈0.001, 0.01）. Packed cell volume presented insignificant difference among groups （P 〉 0.05）. And the erythrocyte aggregation index was significantly lower in treatment group than in model control group and saline control group （0.43±0.04, 0.50±0.05, 0.54±0.04, P 〈 0,05, 0.01）. CONCLUSION： Abnormality of hemorheology can be lead to avascular necrosis of femoral head induced by glucocortieoid. The Chinese herbs of activating blood circulation to remove stasis can strengthen indirectly the expression of VEGF and ameliorate microcireulation of femoral head in the prevention of hormone.