补肾活血中药结合金葡液局部注射治疗骨质疏松性骨折的实验研究

Experimental Research of Bushen Huoxue Herbs Combined with Staphylococcal Enterotoxin C Injection for Repairing Osteoporotic Fracture

目的:研究补肾活血中药结合金葡液注射治疗兔骨质疏松性骨折,分析疗效,探讨作用机制。方法:取雌性兔88只,采用切除卵巢法制作OP模型,随后制作右侧桡骨OPF模型,随机分为4组,每组20只:补肾活血中药结合金葡液治疗组(A组)、补肾活血中药治疗组(B组)、金葡液局部注射治疗组(C组)、骨折未治疗组(D组)。分四个时间段取标本,进行X线摄片、骨痂CT值的测定、HE染色观察、FN和PCNA免疫组化指标的测定,对数据进行统计学分析。结果:X线观察显示不同时间段A组的骨折愈合均较B、C、D组好,结果呈现A>C>B>D排列。CT值测定显示四组骨痂不同时间段CT值均呈递增趋势,8周取材时A组所测的CT值已接近正常骨组织,CT值结果呈现A>C>B>D排列。HE染色显示2、4周时A组染色细胞丰富,排列有序,C、B组次之,6周时A、B、C组骨痂开始成熟,软骨细胞、成骨细胞减少,破骨细胞增加,8周时A组骨小梁致密,排列有序,骨痂开始塑形,以破骨细胞为主,B、C次之,而D组仍有较多软骨细胞、成骨细胞,骨小梁也较稀疏。免疫组化指标结果显示A、B、C组骨痂中FN和PCNA染色阳性细胞初期(2、4周)均表达活跃,且均高于D组,随着骨折愈合进程的发展,表达逐渐减少,8周取材时A、B、C组已不能见到染色阳性细胞,而D组仍有少量表达,整个进程中A组初期表达最活跃,减幅也最显著。结论:补肾活血中药结合金葡液注射能明显促进兔骨质疏松性骨折初期骨痂中FN和PCNA阳性细胞的表达,增加骨痂含量,加快软骨内化骨,改善骨小梁结构,缩短骨折的愈合时间,比单纯补肾活血中药或金葡液注射疗效更好。

Objective ： This research was designed to evaluate the effect of Bushen Huoxue herbs combined with staphylococcal enterotoxin C injection （SECI） for repairing osteoporotic fracture （ OPF）, and to investigate the mechanism of action. Methods ： 80 female rabbits were chosen to make model of OP which were ovanriectomized, then their right radius were made closely fracture for OPF models and were randomized into 4 groups,20 rabbits in each group：group A （treated by Bushen Huoxue herbs combined with SECI） , group B （injected local by SECI only） , group C （treated by Bushen Huoxue herbs only）, group D as the control group. The rabbits were sacrificed in the 4 different periods （2t~ week,4~h week , 6th week ,8th week）, and their right radius were taken for radiography, osteotylus anylysis by CT, hematoxylin and eosin （HE） , FN, the immunohistochemistry index proliferating cell nuclear antigen （PCNA）. All the date was calculated and statistically analyzed. Results：X ray observation showed：the healing in group A in different times was better than that in group B, C and D with the order of A 〉 C 〉 B 〉 D. CT test showed ： the callus at different times in four groups had an in- creasing trend. At the 8th week,the CT result in group A was close to the normal level,with the order of A 〉 C 〉 B 〉 D. HE staining resuhs showed：there were rich cells stained and ordered in group A at 2^nd and 4th week ,followed by group C and B. at 6th week ,the callus in group A, B and C became mature and cartilage cells as well as osteoplast were reduced while osteoelast was increased. At 8th week, the moulding in group A appeared, followed by group B and C. In group D, there were still many cartilage cells and osteoplast. Immunohistochemical indicators showed that FN and PCNA were active in the early period of staining in group A, B and C and were higher than that in group D. As the healing development, FN and PCNA were reduced. At 8th week, there was no staining ceils in group A, B and C. However, in group D, there was still a few expression. Through the whole process, the expression in group A at early stage was active and the reduce was also obvious. Conclusion： Bushen Huoxue herbs combined with staphylococcal enterotoxin C injection can improve the positive cell expressions of FN and PCNA in the early stage of osteoporotic fracture, enrich the quantity of callus,improve the trabecular bone structure and shorten the healing time, which has a better effect than single administration of Bushen Huoxue herbs or staphylococcal enterotoxin C injection.