活血化瘀汤对股骨骨折大鼠骨痂组织胶原表达水平及促进愈合机制分析

Effect of Huoxue Huayu Decoction on Collagen Expression in Callus of Femur Fracture Rat and Its Mechanism of Promoting Healing

目的:探讨活血化瘀汤对股骨骨折大鼠骨痂组织胶原表达水平及促进愈合机制。方法:选取由安康市人民医院和陕西省人民医院实验动物中心提供清洁级雄性SD大鼠43只,依据随机数字表法将大鼠分成两组,观察组(22只)、对照组(21只),股骨中段使用线锯切断制备股骨骨折模型,造模后第二天观察组活血化瘀汤灌胃,对照组使用同剂量生理盐水灌胃;灌胃四周后使用颈椎脱臼法将大鼠处死,察看股骨骨痂生长和骨折愈合状况,同时行HE染色察看大鼠骨痂组织结构,免疫组化检测大鼠治疗后7天、14天、21及28天骨痂组织内Ⅰ与Ⅱ型胶原表达状况;比色法检测血清磷(P)、钙(Ca)、碱性磷酸酶(ALP)含量,ELISA检测血清生长激素(GH)含量。结果:治疗28天时观察组大鼠骨痂组织内钙化状况和骨小梁结构则显著优于对照组,治疗后7天、14天、21天、28天观察组大鼠骨痂组织内Ⅰ型及Ⅱ型胶原表达水平均高于对照组,差异有统计学意义(P<0. 05);治疗后21天、28天观察组大鼠血清GH、P、Ca及ALP水平均高于对照组,差异有统计学意义(P<0. 05)。结论:骨折前期活血化瘀汤可加速骨痂组织内Ⅰ型与Ⅱ型胶原合成,更有利于纤维-软骨性骨痂产生。

Objective:To analyze the expression of collagen in callus tissue of rats with femoral fracture and the mechanism of promoting healing.Methods:Forty-three male SD rats were divided into two groups according to the random number table,the observation group(n=22)and the controlled group(n=21).Preparation of femoral fracture models with cut off,the observation group was given Huoxue Huayu Decoction at the second day with gavage,the controlled group used the same dose of normal saline gavage,four weeks after the use of cervical dislocation method,the rats were sacrificed to observe the femoral callus growth and at the same time,the callus tissue of rat was observed by HE staining.The expression of type I and type II collagen in callus were detected by immunohistochemistry at 7,14,21 and 28 days after treatment.Serum phosphorus(P),calcium(Ca)and alkaline phosphatase(ALP)levels were detected by colorimetric assay.Serum growth hormone(GH)levels were measured by ELISA.Results:At the 28 th day of treatment,calcification and trabecular structure in the callus tissue of the observation group were significantly better than that of the controlled group.At 7,14,21 and 28 days after treatment,the typeⅠand typeⅡCollagen expression levels were higher than the controlled group,the difference was statistically significant(P<0.05).The levels of serum GH,P,Ca and ALP in the observation group were higher than those in the controlled group 21 and 28 days after treatment(P<0.05).Conclusion:Huoxue Huayu Decoction can accelerate the synthesis of type I and type II collagen in callus tissue,and is more conducive to the production of fiber-cartilage callus.