中药骨痛仙胶囊对骨折家兔Ⅰ、Ⅱ型胶原蛋白及BMP-2mRNA表达影响的实验研究

目的:观察骨痛仙胶囊对骨折愈合过程中Ⅰ、Ⅱ型胶原及BMP-2mRNA的转录水平的影响,分析其促进骨折愈合的分子机理。方法:健康成年家兔45只,做双侧桡骨骨折模型,随机分成治疗组、阳性对照组、模型组分别给予骨痛仙胶囊、骨折创伤散和生理盐水灌胃给药,术后1周、2周、4周后处死家兔取材,用原位杂交法测定骨Ⅰ、Ⅱ型胶原及BMP-2表达,并与模型组、阳性对照组比较进行病理学观察。结果:各时间段治疗组Ⅰ型胶原表达mRNA的转录水平均增强,明显高于阳性对照组和模型组(P<0.05)。与对照组和模型组相比,给药组Ⅱ型胶原表达mRNA的转录水平第1周明显提高,第2、4周Ⅱ型胶原明显衰落,与模型组、阳性对照组相比有统计学意义(P<0.05)。给药组骨折部位骨形态发生蛋白-2(BMP-2)mRNA的转录水平比其它二组明显提高(P<0.05),同样在一周时提前达到高峰。结论:本课题从Ⅰ、Ⅱ型胶原及BMP-2mRNA转录三个层次讨论骨痛仙对骨折愈合的影响,从实验结果中我们看到,本实验采用家兔骨折模型,从组织学看,骨折2周达软骨修复高峰,之后进入骨基质骨化及塑性期,直至4周达到塑性的高峰。本实验发现Ⅰ型胶原mR-NA表达在骨折后两周明显小于4周,而Ⅱ型胶原mRNA表达在骨折后2周明显大于4周,这提示Ⅰ型胶原mRNA表达在骨化及塑性期最明显,Ⅱ型胶原mRNA表达在软骨修复期最明显,与Hiltrunen和Sandberg等报道结果一致。若骨痛仙组与阳性对照组、模型组进行比较观察,则发现Ⅰ型胶原mRNA表达,无论在骨折后2周还是4周,前者均大于后两者;而Ⅱ型胶原mRNA表达,前者均小于后两者,提示骨痛仙组的骨形成加快,已提前进入骨化及塑性期。另外,在骨折愈合的不同时期,BMP-2的表达及其mRNA的转录水平呈动态变化,即骨折后开始升高,2周左右达到高峰,以后逐渐下降。1骨痛仙胶囊可明显地促进实验性骨折家兔的骨折愈合,缩短骨折愈合的时间。2骨痛仙胶囊能够调节实验性骨折家兔Ⅰ、Ⅱ型胶原及BMP-2mRNA的转录水平。促进骨痂由纤维骨痂到骨性骨痂的转变。3Ⅰ、Ⅱ型胶原及BMP-2是观察骨折愈合的一项重要指标,在骨折愈合过程中,由软骨组织向骨组织变化过程对应着Ⅱ型胶原向Ⅰ型胶原转变的过程。

Objective:To investigate effect of fraction of GTX capsule on collagen typeⅠtypeⅡand BMP-2 mRNA gene expression in the process of fracture union, and to explore the mechanism of GTX inenhancing fracture healing. Methods:Standardized radial factures were performed in New Zeland White rabbits, which were randomized into three groups:the GTX capsule group, the GZCSS group(the positive control group), and the blank control model group. They were fed orally with GTX, GZCSS, and normal saline respectively, and were sacrificed on 1st week, 2nd week and 4th week after fracture in batches. The femur head was taken or detecting expression of collagen type Ⅰ.Ⅱand BMP-2 mRNA with situ hybridization and pathologic examination. All data were compared with that of the positive control group and the blank control model group.Results: On 1st week, 2nd week and 4th week after fracture , The collagen typeⅠmRNA expression of the GTX capsule group enhance without exception. And is obviously superior to the positive control group and the blank control model group (P<0.05). Compared with the positive control group and the blank control model group, the collagen typeⅡmRNA expression of the GTX capsule group rises clearly on 1st week,but declines on 2nd week and 4th week (P<0.05). Expression of BMP-2 at fracture region in treating group is obviously supeior to the other two groups(P<0.05) and advancely reach peak valuce at first weak. Conclusion:The GTX capsule and the GZCSS can both adjust the expression of collagen concerned in the process of fracture union,and promote fracture union;The GTX capsule is superior than the positive control group and the blank control model group in the expression of collagen type Ⅰ Ⅱand BMP-2 mRNA