生脉成骨胶囊治疗激素性股骨头坏死的骨修复机理研究
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摘要
股骨头坏死是由于不同病因破坏股骨头血供所造成的最终结果，其基本
病理变化是骨细胞、骨小梁坏死，同时伴有血管及骨髓细胞的坏死改变，在坏
死发生的同时，骨组织交错着进行自然修复反应。目前认为激素诱导的股骨头
坏死多为广泛坏死，其修复晚且能力差，成为治疗难题。国内外学者一直在寻
找能真正用于促进坏死修复的有效方法。
本文总结近年来激素性股骨头坏死的研究进展及导师多年临床经验，认为
本病的病机特点是气血不通、气血瘀滞而产生的“瘀血”。生脉成骨胶囊是导
师研制的治疗股骨头坏死的经验方，具有活血化瘀、补肾健骨、消肿止痛之功
效。多年的研究结果表明，生脉成骨胶囊有比较肯定的促进坏死修复的作用，
但作用机理有待深入研究。
本课题采用激素+内毒素注射造成家兔激素性股骨头坏死模型，从光镜、
电镜、原位杂交、骨组织生化代谢、血清骨钙素和IL-6的放射免疫分析等方
面比较深入地研究生脉成骨胶囊促进骨修复的作用，旨在进一步阐明该药的作
用机理，同时也为该药扩大应用范围如治疗骨质疏松症、骨折等骨科疾病提供
理论依据。
成年新西兰大白兔56只，随机选取18只为正常对照组，余38只用于造
模；予兔耳缘静脉注射内毒素每次50ug/kg，共2次，前后间隔24小时，在
第2次注射内毒素后随即腹腔注射甲基强的松龙每次20mg/kg/日，连续三日。
于2周后随机处死动物2只观察组织学变化，确认模型成立后将动物随机分为
2组：模型组、生脉成骨胶囊治疗组。生脉成骨胶囊治疗组用含生药0.42g/ml
药液按5ml/kg体重/日灌胃，模型组和正常组用等量生理盐水灌胃，三组动物
分别于用药后4周、8周、12周各处死6只行各项检查。采血，分离血清，测
定骨钙素和IL-6。取出双侧股骨头行组织病理学、超微结构观察和Ⅰ型胶原mRNA
生脉成骨胶囊治疗激素性股骨头坏死的骨修复机理研究
结构观察和 1型胶原栅NA的原位表达。也采用激素＋内毒素注射法造模，观
察生脉成骨胶囊治疗股骨头坏死后第8周时兔股骨头局部生化代谢的改变。
结果出现：1．光镜观察见模型组空骨陷窝率逐渐升高，成骨细胞计数。
骨小梁相对体积和平均宽度逐渐减少，新骨形成少，用生脉成骨胶囊治疗后
空骨陷窝率减少，成骨细胞计数、骨小梁相对体积和平均宽度逐渐增加，新
骨形成活跃；
2．电镜观察见模型组成骨细胞细胞器损伤严重，骨细胞脂肪变性坏死。
用生脉成骨胶囊治疗后成骨细胞细胞器有修复，骨细胞退变相少；
3，原位杂交见模型组成骨细胞、骨髓前成骨细胞1型胶原帆NA的表达
减少，用生脉成骨胶囊治疗后1型胶原mRNA表达的强度及面积增加；
4．生化分析见模型组软骨下骨胶原、钙磷含量减少，用生脉成骨胶囊治
疗后胶原、钙磷含量升高；
5、放射免疫分析见模型组血清骨钙素水平逐步下降，K－6水平升高，用
生脉成骨胶囊治疗后骨钙素水平上升，1卜6维持在正常水平。
结论：激素性股骨头坏死的骨修复能力有限，骨形成减少，骨丢失增加
与激素性股骨头坏死的病理发展密切相关。生脉成骨胶囊能促进骨形成，抑
制骨丢失，逆转骨坏死的发展。中药促进骨修复的能力随作用时间的延长而
更为明显，其作用机理表现为：①调节骨细胞代谢，抑制骨细胞的变性坏死，
保持骨小梁的完整结构；②促进成骨细胞的增殖分化，提高成骨细胞的活性，
增加 l型胶原InNA的表达和骨钙素的分泌，促进新骨形成；③增加骨胶原
的合成与钙磷的沉积，减少骨丢失；④降低R－6水平，抑制其介导的骨吸
收。
Avascular necrosis of the femoral head (ANFH) results from the blood
supply impairment by various causative factors. Its basic pathologic change
is the death of bone and marrow elements. At the same time, a process of
repair then is initiated. The reparative reaction is late and ineffective in
glueocorticoid-induced avascular necrosis of the femoral head (GANFH),
which is involved extensive size and difficult to treat. Scholars in the world
are looking for effective methods in improving the reparative process.
GANFH belongs to blood stasis in TCM. Shengnai Chenggu Capsule is a
recipe made by Prof. Yuan llao for the treatment on avascular necrosis of the
femoral head. It is efficacious in activating circulation of the blood so as to
eleminate blood stasis, tonifying the kidney and strengthening the bone,
suppressing inflammation and alleviating pains. The long-term clinical
practice has proved it is an effective medicine for ANFH. But its effect
mechanism remains further research.
GANFH model in rabbits was induced by injecting endotoxin and
niethylprednisolone. We investigated the promotion effect mechanism of
Shengmai Chenggu Capsule on GANFII.
56 New Zealand rabbits were randomly allocated into normal control
group and the test. 38 rabbits in the test were given S0uglkg of endotoxin
from Escherichia intrvenously twice at an interval of 24 hours, and injected
intramuscularly with 20mg/kg of methylpreduisolone 3 times at intervals of
24 hours. 2 rabbits were sacrificed for ascertainment of GANFLI after two
weeks. The left were randomly allocated into model group and treatment
gr1u1 With Shengmai Chenggu Capsule. Rabbits in treatment group were
respectively received Shengmai Chenggu Capsule 2.lg.k?d? Rabbits in
normal and model control group were respectively received 2.lg.k?d?Saline.
6 rabbits were killed at the time of fourth week, eighth week, twelfth week in
each group respectively. Bone gla protein (BGP) and interleukin-6 (IL-6) in
serum were assayed.
Results :1. By light microscopy, in model group empty lacunae
increased. Quality of osteoblasts, MTW and TBV in model group decreased.
Specimens in treatment group showed those pathologic changes relieved
gradually along with active new bone formation.
2. By electron microscopy, in model group the function organeli in
osteoblasts showed impaired severely. Small lipid droplets in osteocytes
gradually, which resulted in cell death, in treatment group organeli in
osteoblasts and osteocytes maintained intact structure.
3. In situ hybridization, it decreased expression of type I collagen
messenger RNA in osteoblasts and osteoprogenitor cells in model group,
whereas increased in treatment group.
4 By biochemistry analysis, it decreased the bone content of collagen,
calcium and phosphorus in subchondral bone in model group. It increased in
treatment group.
5. By radioimmunology analysis, level of BGP in sum gradually
decreased in model group, whereas LL-6 increased. Level of BGP in serum
gradually increased in treatment group, whereas IL-6 maintained normal
level.
In conclusion, decreased bone formation and increased bone loss awe
close correlated to disease progression of GANFII. Shengn~qi Che~t鏶u
Capsule is able to reverse disease progression With improving bone form~ion
and inhibiting bone loss. This effect is more obvious in a time-dependent
mann
病理变化是骨细胞、骨小梁坏死，同时伴有血管及骨髓细胞的坏死改变，在坏
死发生的同时，骨组织交错着进行自然修复反应。目前认为激素诱导的股骨头
坏死多为广泛坏死，其修复晚且能力差，成为治疗难题。国内外学者一直在寻
找能真正用于促进坏死修复的有效方法。
本文总结近年来激素性股骨头坏死的研究进展及导师多年临床经验，认为
本病的病机特点是气血不通、气血瘀滞而产生的“瘀血”。生脉成骨胶囊是导
师研制的治疗股骨头坏死的经验方，具有活血化瘀、补肾健骨、消肿止痛之功
效。多年的研究结果表明，生脉成骨胶囊有比较肯定的促进坏死修复的作用，
但作用机理有待深入研究。
本课题采用激素+内毒素注射造成家兔激素性股骨头坏死模型，从光镜、
电镜、原位杂交、骨组织生化代谢、血清骨钙素和IL-6的放射免疫分析等方
面比较深入地研究生脉成骨胶囊促进骨修复的作用，旨在进一步阐明该药的作
用机理，同时也为该药扩大应用范围如治疗骨质疏松症、骨折等骨科疾病提供
理论依据。
成年新西兰大白兔56只，随机选取18只为正常对照组，余38只用于造
模；予兔耳缘静脉注射内毒素每次50ug/kg，共2次，前后间隔24小时，在
第2次注射内毒素后随即腹腔注射甲基强的松龙每次20mg/kg/日，连续三日。
于2周后随机处死动物2只观察组织学变化，确认模型成立后将动物随机分为
2组：模型组、生脉成骨胶囊治疗组。生脉成骨胶囊治疗组用含生药0.42g/ml
药液按5ml/kg体重/日灌胃，模型组和正常组用等量生理盐水灌胃，三组动物
分别于用药后4周、8周、12周各处死6只行各项检查。采血，分离血清，测
定骨钙素和IL-6。取出双侧股骨头行组织病理学、超微结构观察和Ⅰ型胶原mRNA
生脉成骨胶囊治疗激素性股骨头坏死的骨修复机理研究
结构观察和 1型胶原栅NA的原位表达。也采用激素＋内毒素注射法造模，观
察生脉成骨胶囊治疗股骨头坏死后第8周时兔股骨头局部生化代谢的改变。
结果出现：1．光镜观察见模型组空骨陷窝率逐渐升高，成骨细胞计数。
骨小梁相对体积和平均宽度逐渐减少，新骨形成少，用生脉成骨胶囊治疗后
空骨陷窝率减少，成骨细胞计数、骨小梁相对体积和平均宽度逐渐增加，新
骨形成活跃；
2．电镜观察见模型组成骨细胞细胞器损伤严重，骨细胞脂肪变性坏死。
用生脉成骨胶囊治疗后成骨细胞细胞器有修复，骨细胞退变相少；
3，原位杂交见模型组成骨细胞、骨髓前成骨细胞1型胶原帆NA的表达
减少，用生脉成骨胶囊治疗后1型胶原mRNA表达的强度及面积增加；
4．生化分析见模型组软骨下骨胶原、钙磷含量减少，用生脉成骨胶囊治
疗后胶原、钙磷含量升高；
5、放射免疫分析见模型组血清骨钙素水平逐步下降，K－6水平升高，用
生脉成骨胶囊治疗后骨钙素水平上升，1卜6维持在正常水平。
结论：激素性股骨头坏死的骨修复能力有限，骨形成减少，骨丢失增加
与激素性股骨头坏死的病理发展密切相关。生脉成骨胶囊能促进骨形成，抑
制骨丢失，逆转骨坏死的发展。中药促进骨修复的能力随作用时间的延长而
更为明显，其作用机理表现为：①调节骨细胞代谢，抑制骨细胞的变性坏死，
保持骨小梁的完整结构；②促进成骨细胞的增殖分化，提高成骨细胞的活性，
增加 l型胶原InNA的表达和骨钙素的分泌，促进新骨形成；③增加骨胶原
的合成与钙磷的沉积，减少骨丢失；④降低R－6水平，抑制其介导的骨吸
收。
Avascular necrosis of the femoral head (ANFH) results from the blood
supply impairment by various causative factors. Its basic pathologic change
is the death of bone and marrow elements. At the same time, a process of
repair then is initiated. The reparative reaction is late and ineffective in
glueocorticoid-induced avascular necrosis of the femoral head (GANFH),
which is involved extensive size and difficult to treat. Scholars in the world
are looking for effective methods in improving the reparative process.
GANFH belongs to blood stasis in TCM. Shengnai Chenggu Capsule is a
recipe made by Prof. Yuan llao for the treatment on avascular necrosis of the
femoral head. It is efficacious in activating circulation of the blood so as to
eleminate blood stasis, tonifying the kidney and strengthening the bone,
suppressing inflammation and alleviating pains. The long-term clinical
practice has proved it is an effective medicine for ANFH. But its effect
mechanism remains further research.
GANFH model in rabbits was induced by injecting endotoxin and
niethylprednisolone. We investigated the promotion effect mechanism of
Shengmai Chenggu Capsule on GANFII.
56 New Zealand rabbits were randomly allocated into normal control
group and the test. 38 rabbits in the test were given S0uglkg of endotoxin
from Escherichia intrvenously twice at an interval of 24 hours, and injected
intramuscularly with 20mg/kg of methylpreduisolone 3 times at intervals of
24 hours. 2 rabbits were sacrificed for ascertainment of GANFLI after two
weeks. The left were randomly allocated into model group and treatment
gr1u1 With Shengmai Chenggu Capsule. Rabbits in treatment group were
respectively received Shengmai Chenggu Capsule 2.lg.k?d? Rabbits in
normal and model control group were respectively received 2.lg.k?d?Saline.
6 rabbits were killed at the time of fourth week, eighth week, twelfth week in
each group respectively. Bone gla protein (BGP) and interleukin-6 (IL-6) in
serum were assayed.
Results :1. By light microscopy, in model group empty lacunae
increased. Quality of osteoblasts, MTW and TBV in model group decreased.
Specimens in treatment group showed those pathologic changes relieved
gradually along with active new bone formation.
2. By electron microscopy, in model group the function organeli in
osteoblasts showed impaired severely. Small lipid droplets in osteocytes
gradually, which resulted in cell death, in treatment group organeli in
osteoblasts and osteocytes maintained intact structure.
3. In situ hybridization, it decreased expression of type I collagen
messenger RNA in osteoblasts and osteoprogenitor cells in model group,
whereas increased in treatment group.
4 By biochemistry analysis, it decreased the bone content of collagen,
calcium and phosphorus in subchondral bone in model group. It increased in
treatment group.
5. By radioimmunology analysis, level of BGP in sum gradually
decreased in model group, whereas LL-6 increased. Level of BGP in serum
gradually increased in treatment group, whereas IL-6 maintained normal
level.
In conclusion, decreased bone formation and increased bone loss awe
close correlated to disease progression of GANFII. Shengn~qi Che~t鏶u
Capsule is able to reverse disease progression With improving bone form~ion
and inhibiting bone loss. This effect is more obvious in a time-dependent
mann
引文
1 胥少汀，葛宝奉，徐印坎主编．实用骨科学．人民军医出版社，1999，第二版 P1072-1076
2 刘培林，夏仁云，徐静．激素对股骨头骨细胞的损害．骨与关节损伤，2000，15(5)：397-398
3 Kawai K, Tamaki A, Hirohata K. Steroid-induced accumulation of lipid in the osteocytes of the rabbit femoral head: a histological electron microscopic study. J Bone Joint Surg(Am), 1985, 67: 755-763.
4 Uno K, Kawai K, Hirohata K, et al. Steroid induced early changes of the femoral head in man-histological study of autopsied cases. Ryumachi, 1991, 31: 282-289.
5 Humphreys S, Spencer JD, Tighe JR. The femoral head in osteonecrosis: a quantitative study of osteoeyte population. J Bone Joint Surg(Br), 1989, 71: 205-208.
6 袁浩，方斌，何伟，等 生脉成骨胶囊治疗激素性股骨头坏死的实验研究 中医正骨 1999 11(8)：3-4
7 Weinstein RS, Jilka RI, Parfitt AM, et al. Inhibition of osteoblastogenisis and promotion of apotosis of osteoblasts and osteocytes by glucocorticoids. J Clin Invest, 1998, 102: 274-282
8 Doherty WJ, Derome ME, McCarthy MB. The effect of glucocorticoids on osteoblast function. J Bone Joint Surg, 1995, 77A: 396-405.
9 Defranco DJ, Lian JB, Glowacki J. Differential effects of glucocorticoid on recruitment and activity of osteoclasts induced by normal and osteocalcin deficient bone implanted in rats. Endocrinology, 1992, 131(1): 114.
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11 Vidal NO, Brandstrom H, Joneson KB, et al. Osteoprotegerin mRNA is expressed in primary human osteoblast-like cell: dowmregulation by glucocorticoids. J Endocrinol, 1998. 159: 191-195
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19 郑晓辉，方秀斌．骨钙素的基础与应用研究进展．解剖科学进展，1999．5(4)：327-329
20 Bikle DD, Halloran B, Fong L, et al. Elevated 1, 25-dihydroxyvitamin D levels in patients with chronic obstructive pulmonary disease treated with prednisone. J Clin Endocrinol Metab, 1993, 76(2): 456.
21 Morris HA, Need AG, O' Loughlin PD, et al. Malabsorption of calcium in corticosteroid-induced osteoporosis. Calcif Tissue Int, 1990, 46: 305.
22 黄洪，田成功，睾酮与雄性大鼠骨质疏松关系的实验研究．中华内分泌代谢杂志，1999，15(5)：304-307
23 Montecucco C, Caporali R, Caprotfi P, et al. Sex hormones and bone metabolism in postmentopausal rheumatoid arthritis treated with two different glucocorticoids. J Rheumatol, 1992, 19(12): 1895.
24 赵万军，肖鲁伟，童培建，等．右归饮、鹿马片对激素诱导的兔股骨头坏死血浆雌二醇、睾酮的影响及疗效观察．中医正骨 2000，12(1)：3-5
25 刘长安，张卫平，周吉怀，等 激素性股骨头坏死血浆一氧化氮含量的变化．中华实验外科杂志 1998，15(6)：547-548
26 罗南萍，杨道理，付志厚，等TNF-α、HA和LN对股骨头缺血性坏死致病作用的研究．中国骨伤，1999，13(4)：204-206
27 薛元锁，时述山，李亚非，等．激素性股骨头坏死病程中骨形态发生蛋白-2的改变及其意义．中华实验外科杂志 2000，17(5)：455-456
2 刘培林，夏仁云，徐静．激素对股骨头骨细胞的损害．骨与关节损伤，2000，15(5)：397-398
3 Kawai K, Tamaki A, Hirohata K. Steroid-induced accumulation of lipid in the osteocytes of the rabbit femoral head: a histological electron microscopic study. J Bone Joint Surg(Am), 1985, 67: 755-763.
4 Uno K, Kawai K, Hirohata K, et al. Steroid induced early changes of the femoral head in man-histological study of autopsied cases. Ryumachi, 1991, 31: 282-289.
5 Humphreys S, Spencer JD, Tighe JR. The femoral head in osteonecrosis: a quantitative study of osteoeyte population. J Bone Joint Surg(Br), 1989, 71: 205-208.
6 袁浩，方斌，何伟，等 生脉成骨胶囊治疗激素性股骨头坏死的实验研究 中医正骨 1999 11(8)：3-4
7 Weinstein RS, Jilka RI, Parfitt AM, et al. Inhibition of osteoblastogenisis and promotion of apotosis of osteoblasts and osteocytes by glucocorticoids. J Clin Invest, 1998, 102: 274-282
8 Doherty WJ, Derome ME, McCarthy MB. The effect of glucocorticoids on osteoblast function. J Bone Joint Surg, 1995, 77A: 396-405.
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