丹参有效部位对骨折愈合过程中胶原基因表达的影响

Effect of Effective Fraction of Radix Salviae Miltiorrhizae on Procollagen Gene Expression in Fracture Healing

观察丹参有效部位（ＲＳ９４０３）对骨折愈合过程中前胶原、转化生长因子β１（ＴＧＦβ１）基因表达模式的影响，分析ＲＳ９４０３促进骨折愈合作用的分子机理。方法：健康成年Ｗｉｓｔａｒ大鼠２４只，作双侧桡骨骨折模型，随机分成ＲＳ９４０３组、阳性对照药三花接骨散组（三花组）和空白对照组（空白组），分别以ＲＳ９４０３、三花接骨散和生理盐水灌胃给药，术后３天、 １周、 ２周和 ４周后处死大鼠取材，采用不脱钙的大鼠骨痂组织冰冻切片进行原位杂交，观察骨痂组织中前胶原和 ＴＧＦβ１基因的表达。结果：骨折后第 ３天，ＲＳ９４０３组与三花组的骨折端 ＴＧＦβ１ ｍＲＮＡ的表达明显高子空白组，并已出现血型胶原基因的表达。骨折第１周末，成纤维细胞和软骨细胞样细胞的Ⅲ型前胶原基因表达占主导，ＲＳ９４０３组与三花组Ⅱ、Ⅰ型前胶原与 ＴＧＦβ１基因的表达较空白组里同步增强趋势。骨折第 ２周末， Ⅰ型前胶原 ｍＲＮＡ表达明显增加，ＲＳ９４０３组和三花组中肥大软骨细胞明显多于空白组，Ⅱ型前胶原ｍＲＮＡ的表达呈现显著的衰落趋势。同时证实共有表型表达现象的存在，尤以ＲＳ９４０３组与三花组更为明显。骨折第４周末，软骨骨痂基本被骨组织替代。 ＲＳ９４０３组和三花组?

To investigate effect of effective fraction of Radix Salviae Miltiorrhizae (RS9403) on procollagens and transfroming growth factor β1 (TGF β1) gene expression in fracture callus, and to explore the mechanism of RS9403 in enhancing fracture healing. Methods: Standardized radial fractures were performed in 24 Wistar rats, which were randomized into three groups: the RS9403 group, the Sanhua Jiegu powder (SHJGP) group and the blank control model group. They were fed orally with RS9403, SHJGP and normal saline respectively, and were sacrificed on the 3rd day, 1st week, 2nd week and 4th week after fracture in batches. In situ localization of procollagens and TGF β1 gene expression were examined on the cryosection of rat fracture callus. Results: On the 3rd day after fracture, the TGF β1 gene expression at the site of fracture in the RS9403 group and the SHJGP group was significantly higher than that in the blank control group, and type-Ⅲ procollagen gene expression was observed in the two groups. At the end of 1st week after fracture, the proα1 (Ⅲ ) expression in fibroblast and chondrocyte-like cells was dominant. The local type Ⅱ and Ⅰ procollagen gene expressions in the RS9403 and the SWGP group were enhanced simultaneously with TGF gene expression compared with those in the blank control group. At the end of 2nd week the RS9403 and the SHJGP group were characterized by a marked increase in the mRNA levels of type 1 procollagen, the numbers of hypertrophic chondrocytes was larger than that in the control group, and the type Ⅱ procollagen expression declined markedly. The shared phenotype expression was confirmed at this stage, especially in the RS9403 and the SHJGP group. At the end of 4th week, the cartilagi nous callus was almost all replaced by the bone tissue. In the RS9403 and the SHJGP group, the replacement was nearly complete, few type I procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus. Conclusion: RS9403 could affect the procollagens expression to enhance the healing of bone fracture as SHJGP do, maybe, by modulating the TGF β1 expression.