普拉固对动物激素性股骨头坏死的干预研究
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摘要
目的：采用激素（地塞米松，Dexamethasone）制作成兔股骨头坏死(Steroid-induced Femoral Head Necrosis, SIFHN) 模型，探讨普拉固（Pravastatin）对SIFHN干预作用及其机制。
方法：12-18周龄日本大白兔42只，雌雄不限，体重2.50.6kg,随机分为4组：空白组（Group A）：6只。对照组(单纯激素组Group B)：12只。小剂量普拉固组（Group C）：12只，普拉固口服5mg/天。大剂量普拉固组（Group D）：12只，普拉固口服10mg/天，各组动物均以标准饲料及草料喂养，除A组外，B,C,D组均以Dexamethasone 2.5mg/kg·d im，用足12周。期间每组均予青霉素80万u/只， im。链霉素1.0g/只，im，2次/周，防止感染。分别于6周，8周，10周，12周处死每组兔子各3只，处死前均检查血脂，血粘度，血PT,t-PA及双髋关节x-ray摄片，处死后取双股骨头行HE染色，标本行光学电镜观察，及透射电镜检查，细胞凋亡分析。
结果: 普拉固组中小剂量组及大剂量组的兔血脂，血粘度 PT, t-PA较激素组均升高 （P<005）。与单纯激素组比较，普拉固组X-ray 示股骨头及髋臼可见软骨下区不规则囊性透亮区及骨化硬化带明显减少，HE染色示骨小梁未见稀疏，骨陷窝空虚率，髓内基质细胞脂肪转化率减低，髓内血管栓子形成减少。透射电镜示骨细胞吞饮脂滴减少。细胞凋亡分析示实验组较对照组骨细胞凋亡指数减少（P<005）。但普拉固这种对SIFHN的干预作用，大小(10mg/d与5mg/d)剂量间不存在显著差异。
结论: 地塞米松能在兔体内成功造成股骨头坏死模型，而他汀类药物普拉固能有效干预股骨头坏死进展，其作用机制可能与降血脂，血粘度，抗凝，从而缓解股骨头血供障碍，减少骨质疏松避免软骨下骨骨折，降低骨内压有关。
Object : To build a model of rabbits’ Steroid-induced Femoral Head Necrosis (SIFHN) by administing Dexamethasone(DEX). Discussing whether the Provastatine can intervening the course of resulting in SIFHN and the possible mechanism. Methods: The Japanese rabbits, 12-18 weeks old ,2.50.6kg, randomly divided into 4 groups, Group A(n=8): Just used as the control. Group B(purely administering DEX n=12).Group C(n=12): Pravastatin 5mg/d p.o. Group D(n=12): Pravastatin 10mg·d-1 p.o. Every rabbit of group A B C was fed by standard forage and fodder. DEX of 2.5mg·kg-1·d-1 was injeceted to the rabbits except Group A for 12 weeks. Simultaneity, the Penicillin-natrium 80,0000u and streptomycin 1.0g was injected to everyone ,twice a week, avoiding being infected. Every following 6,8,10,12 weeks, 3 rabbits of every group were selected to examined with serum cholesterol and triglyceride, hemorrhelogical factor, PT, tissue plasminogen( t-PA) and double hipcoxa joints were taken an X-ray photograph before they were executed . The specimens of double femoral heads were taken to studied by the light microscope after stained with Hematoxylin and eosin(HE), Transmission Electron Microscope(TEM) and analyse of apoptosis. Result: Comparing with the Group B, the serum cholesterol and triglyceride, hemorrhelogical factor levels of the Group C and D had lowered obviously, PT and t-PA are higher since 6 weeks (P<0.05),X-ray didn’t show the phenomenons of the irregular radiolucent cystic area and ossificational cirrhosis that was found clearly in the subchondral region of the acetabulum and femoral head. Light microscopy exhibit the density of trabecular bone did not change more than Group A. The rate of the empty lacuna and the bone marrow stromal cell transforming to the lipocyte had decreased obviously . There is seldom fat embolus and lipid droplets appearing in the vessels of the osteocytes. Under the TEM, organelle and nucleus of osteocytes were found rarely degenerative, necrotic and swallowing the lipid droplets. The analyse of apoptosis present the AI(Apoptosis Index) was lower than Group B（P<005） .The effect of
Pravastatin intervening the course of SIFHN did not show markedly difference between the Group C and D. Conclusion: The Dexamethasone resulted in building the model of rabbits’ SIFHN as the Group B. and the Pravastatin can intervene the couse of SIFHN effectively .The mechanism of action is mightly related with lowering the level of serum lipid and hemorrhelogical factor, releasing the osteoporosis, which would reduce the obstructive possibility of blood supply and the rate of fracture that happened below the subchondral region of femoral head.
方法：12-18周龄日本大白兔42只，雌雄不限，体重2.50.6kg,随机分为4组：空白组（Group A）：6只。对照组(单纯激素组Group B)：12只。小剂量普拉固组（Group C）：12只，普拉固口服5mg/天。大剂量普拉固组（Group D）：12只，普拉固口服10mg/天，各组动物均以标准饲料及草料喂养，除A组外，B,C,D组均以Dexamethasone 2.5mg/kg·d im，用足12周。期间每组均予青霉素80万u/只， im。链霉素1.0g/只，im，2次/周，防止感染。分别于6周，8周，10周，12周处死每组兔子各3只，处死前均检查血脂，血粘度，血PT,t-PA及双髋关节x-ray摄片，处死后取双股骨头行HE染色，标本行光学电镜观察，及透射电镜检查，细胞凋亡分析。
结果: 普拉固组中小剂量组及大剂量组的兔血脂，血粘度 PT, t-PA较激素组均升高 （P<005）。与单纯激素组比较，普拉固组X-ray 示股骨头及髋臼可见软骨下区不规则囊性透亮区及骨化硬化带明显减少，HE染色示骨小梁未见稀疏，骨陷窝空虚率，髓内基质细胞脂肪转化率减低，髓内血管栓子形成减少。透射电镜示骨细胞吞饮脂滴减少。细胞凋亡分析示实验组较对照组骨细胞凋亡指数减少（P<005）。但普拉固这种对SIFHN的干预作用，大小(10mg/d与5mg/d)剂量间不存在显著差异。
结论: 地塞米松能在兔体内成功造成股骨头坏死模型，而他汀类药物普拉固能有效干预股骨头坏死进展，其作用机制可能与降血脂，血粘度，抗凝，从而缓解股骨头血供障碍，减少骨质疏松避免软骨下骨骨折，降低骨内压有关。
Object : To build a model of rabbits’ Steroid-induced Femoral Head Necrosis (SIFHN) by administing Dexamethasone(DEX). Discussing whether the Provastatine can intervening the course of resulting in SIFHN and the possible mechanism. Methods: The Japanese rabbits, 12-18 weeks old ,2.50.6kg, randomly divided into 4 groups, Group A(n=8): Just used as the control. Group B(purely administering DEX n=12).Group C(n=12): Pravastatin 5mg/d p.o. Group D(n=12): Pravastatin 10mg·d-1 p.o. Every rabbit of group A B C was fed by standard forage and fodder. DEX of 2.5mg·kg-1·d-1 was injeceted to the rabbits except Group A for 12 weeks. Simultaneity, the Penicillin-natrium 80,0000u and streptomycin 1.0g was injected to everyone ,twice a week, avoiding being infected. Every following 6,8,10,12 weeks, 3 rabbits of every group were selected to examined with serum cholesterol and triglyceride, hemorrhelogical factor, PT, tissue plasminogen( t-PA) and double hipcoxa joints were taken an X-ray photograph before they were executed . The specimens of double femoral heads were taken to studied by the light microscope after stained with Hematoxylin and eosin(HE), Transmission Electron Microscope(TEM) and analyse of apoptosis. Result: Comparing with the Group B, the serum cholesterol and triglyceride, hemorrhelogical factor levels of the Group C and D had lowered obviously, PT and t-PA are higher since 6 weeks (P<0.05),X-ray didn’t show the phenomenons of the irregular radiolucent cystic area and ossificational cirrhosis that was found clearly in the subchondral region of the acetabulum and femoral head. Light microscopy exhibit the density of trabecular bone did not change more than Group A. The rate of the empty lacuna and the bone marrow stromal cell transforming to the lipocyte had decreased obviously . There is seldom fat embolus and lipid droplets appearing in the vessels of the osteocytes. Under the TEM, organelle and nucleus of osteocytes were found rarely degenerative, necrotic and swallowing the lipid droplets. The analyse of apoptosis present the AI(Apoptosis Index) was lower than Group B（P<005） .The effect of
Pravastatin intervening the course of SIFHN did not show markedly difference between the Group C and D. Conclusion: The Dexamethasone resulted in building the model of rabbits’ SIFHN as the Group B. and the Pravastatin can intervene the couse of SIFHN effectively .The mechanism of action is mightly related with lowering the level of serum lipid and hemorrhelogical factor, releasing the osteoporosis, which would reduce the obstructive possibility of blood supply and the rate of fracture that happened below the subchondral region of femoral head.
引文
Kawaik,Tamaki A,Hirohata. Sterioid-induced accumulation of lipid in the osteocytos of the rabbits femoral headl. J Bone Joint Surg (Am),1985,67:755-757.
Cui Q,Wang G L,Ba Lian G. Steriod induced adipogenesis in a pluripotential cell line from bone marrow 〔J〕.J Bone Joint Surg (Am),1997,79:1054-1063.
周谋望，秦建中，刘志雄等. 降脂药物防治激素所致股骨头骨细胞损害的实验研究.中华医学杂志，1996,76(1):13-16.
贺西京.肾上腺皮质激素引起股骨头缺血坏死的机制实验研究.中华骨科杂志，1992，12：440
Wang GJ.et al. Fat-cellchanges as a mechanism of avascular necrosis of the fermoral head in cortisone-treated rabbits. J Bone Joint Surg, 1977, 59A:729.
王坤正，毛履真，胡长根，等.激素性股骨头缺血坏死发病机制的实验研究. 中华实验外科杂志， 10994, 32:515
刘培林，夏仁云，张士杰. 股骨头缺血性造模的实验研究. 中国矫形外科杂志，2001,8:889
刘忠堂. 酒精和激素性股骨头缺血性坏死发病机制的实验研究. 温州医学院报，1999，29：184.
周强.糖皮质激素诱导性股骨头坏死模型的血管改变.中华外科杂志，2000，38：212
同志勤.实验性股骨头坏死免疫组织化学与病理改变研究,西安医科大学学报， 1998：452
洪加源.激素性股骨头坏死坏死骨代谢变化的实验研究.骨与关节损伤杂志,2001,16:365
曾晓峰，赵建宁.股骨头缺血性坏死病理改变及早期诊断.人民军医, 1998,41:207
Wang GJ, et al.The effect of core decompression on fermoral head blood flow in steroid-induced avascular necrosis of the fermoral head .J Bone Joint Surg(AM) 1985, 67:121.
Caffey J, Paper A. Avascular necrosis: early MR imaging and histological
findings in a canine model. AJR,1991,157：341～345
王坤正，杨万石，王春生等.特发性股骨头缺血坏死早期细胞超微结构改变的实验研究.中华矫形外科杂志,1996，1：51.
Alert J. Nontraumatic avascular necrosis of the femoral head. Clin orthrop,1992,227:12-21.
Kisker C T,robillad JE,Boh iken Dp . Glucocirticoid Stimulation of blood coagulation factor activitios in the fetal lamb.J J Lab Clin Med ,1983,101:509.
Darrell E,William H. Corticosteroid-inducced avascular necrosis . J Bone Joint Surg(Am),1971,53:859.
Jones JP.Intravascular coagulation and osteonecrosis. Clin Orthop, 1992, 2777:98
Kawaik,Tamaki A,Hirohata. Sterioid-induced accumulation of lipid in the osteocytos of the rabbits femoral headl. J Bone Joint Surg (Am),1985,67:755-757.
Kawai K,Maruno H,Fat Necrosis of osteonecrosis in heritable hyperlipaemic rabbits. Clin Orthop,1980,153:273.
周谋望，秦建中，朱盛雄，激素导致股骨头骨细胞损害的研究. 中华实验外科杂志， 1994，11：89
Arlet J. Ischemia and necrosis of bone. Baltimere l London, Williams & wilkins, 1980,29
付志厚，谭项玲，王本堂，等.股骨头缺血性坏死患者的实验室检查结果分析.山东医药，1995，5：20～21
李喜东，范力军，李国力，等. 介入治疗激素性股骨头坏死的.中华放射学杂志，1998，1：32～35
Rosenson RS,Tangney CC. Antibther othrombotic properties of statins:implications for Cardiovascular event reduction. JAMA,1998,279:1043-1650
Aoki I,Aokin, Kawanok,et al. Platelet-dependent thrombin generation in pationts with by perlipidaemia. J Am Coll Cardiol,1997,30:91-96.
okatori Y. Avascular necrosis of the femoral head: natural history and
magnetic resonance imaging. J Bone Joint Surg,1993,75B∶217～221
Dangas G,Badimon JJ,Smith DA, et al. Pravastatin therapy in hyperlipidemia:effects on thrombus formation and systemic hemostatic profile. J Am Coll Cardiol,1999,33(5):1294-1304.
Wyllie AH. The genetic regulation of apoptosis. Curr Opin Genet Dev, 1995, 5: 97-104.
罗义 李卓仁 郭南山 心血管疾病中的细胞凋亡 1997-02-03
Robert S. Weinstein, Robert L,et al. Inhibition of Osteoblastogenesis and Promotion of Apoptosis of Osteoblasts and Osteocytes by Glucocorticoids.J. Clin. Invest, Number, July 1998, 102(2)274-282.
Cui Q,Wang G L,Ba Lian G. Steriod induced adipogenesis in a pluripotential cell line from bone marrow 〔J〕.J Bone Joint Surg (Am),1997,79:1054-1063.
周谋望，秦建中，刘志雄等. 降脂药物防治激素所致股骨头骨细胞损害的实验研究.中华医学杂志，1996,76(1):13-16.
贺西京.肾上腺皮质激素引起股骨头缺血坏死的机制实验研究.中华骨科杂志，1992，12：440
Wang GJ.et al. Fat-cellchanges as a mechanism of avascular necrosis of the fermoral head in cortisone-treated rabbits. J Bone Joint Surg, 1977, 59A:729.
王坤正，毛履真，胡长根，等.激素性股骨头缺血坏死发病机制的实验研究. 中华实验外科杂志， 10994, 32:515
刘培林，夏仁云，张士杰. 股骨头缺血性造模的实验研究. 中国矫形外科杂志，2001,8:889
刘忠堂. 酒精和激素性股骨头缺血性坏死发病机制的实验研究. 温州医学院报，1999，29：184.
周强.糖皮质激素诱导性股骨头坏死模型的血管改变.中华外科杂志，2000，38：212
同志勤.实验性股骨头坏死免疫组织化学与病理改变研究,西安医科大学学报， 1998：452
洪加源.激素性股骨头坏死坏死骨代谢变化的实验研究.骨与关节损伤杂志,2001,16:365
曾晓峰，赵建宁.股骨头缺血性坏死病理改变及早期诊断.人民军医, 1998,41:207
Wang GJ, et al.The effect of core decompression on fermoral head blood flow in steroid-induced avascular necrosis of the fermoral head .J Bone Joint Surg(AM) 1985, 67:121.
Caffey J, Paper A. Avascular necrosis: early MR imaging and histological
findings in a canine model. AJR,1991,157：341～345
王坤正，杨万石，王春生等.特发性股骨头缺血坏死早期细胞超微结构改变的实验研究.中华矫形外科杂志,1996，1：51.
Alert J. Nontraumatic avascular necrosis of the femoral head. Clin orthrop,1992,227:12-21.
Kisker C T,robillad JE,Boh iken Dp . Glucocirticoid Stimulation of blood coagulation factor activitios in the fetal lamb.J J Lab Clin Med ,1983,101:509.
Darrell E,William H. Corticosteroid-inducced avascular necrosis . J Bone Joint Surg(Am),1971,53:859.
Jones JP.Intravascular coagulation and osteonecrosis. Clin Orthop, 1992, 2777:98
Kawaik,Tamaki A,Hirohata. Sterioid-induced accumulation of lipid in the osteocytos of the rabbits femoral headl. J Bone Joint Surg (Am),1985,67:755-757.
Kawai K,Maruno H,Fat Necrosis of osteonecrosis in heritable hyperlipaemic rabbits. Clin Orthop,1980,153:273.
周谋望，秦建中，朱盛雄，激素导致股骨头骨细胞损害的研究. 中华实验外科杂志， 1994，11：89
Arlet J. Ischemia and necrosis of bone. Baltimere l London, Williams & wilkins, 1980,29
付志厚，谭项玲，王本堂，等.股骨头缺血性坏死患者的实验室检查结果分析.山东医药，1995，5：20～21
李喜东，范力军，李国力，等. 介入治疗激素性股骨头坏死的.中华放射学杂志，1998，1：32～35
Rosenson RS,Tangney CC. Antibther othrombotic properties of statins:implications for Cardiovascular event reduction. JAMA,1998,279:1043-1650
Aoki I,Aokin, Kawanok,et al. Platelet-dependent thrombin generation in pationts with by perlipidaemia. J Am Coll Cardiol,1997,30:91-96.
okatori Y. Avascular necrosis of the femoral head: natural history and
magnetic resonance imaging. J Bone Joint Surg,1993,75B∶217～221
Dangas G,Badimon JJ,Smith DA, et al. Pravastatin therapy in hyperlipidemia:effects on thrombus formation and systemic hemostatic profile. J Am Coll Cardiol,1999,33(5):1294-1304.
Wyllie AH. The genetic regulation of apoptosis. Curr Opin Genet Dev, 1995, 5: 97-104.
罗义 李卓仁 郭南山 心血管疾病中的细胞凋亡 1997-02-03
Robert S. Weinstein, Robert L,et al. Inhibition of Osteoblastogenesis and Promotion of Apoptosis of Osteoblasts and Osteocytes by Glucocorticoids.J. Clin. Invest, Number, July 1998, 102(2)274-282.