| Objective: Observe the effects of Huo Xue Hua Yu (HXHY) decoction on serum bonegamma Carboxyglutamate protein (BGP), Collagen I, transforming growth factor-β1(TGF-β1), genetic expression of callus of fracture sides at level of mRNA and protein, in ord-er to explicate the endochondral ossification mechanism during fracture healing via paraffin s-ection, electron mi-croscoe of transmission, X-ray, anti-fracture efficacy.Methods: Standardized fracture model of the right tibia in 99 anesthetized male Sprague-Dawley rats with a weight of 280±30g was induced using a modified technique. Fraturesides were fixed by percutaneous intramedullary nails (84 rats up to commamd), 6 normalrats served as control group. All rats were filled with HXHY decoction served as treatmentgroup while normal sodium served as control groups after fracture. Blood serum wasdetermined by the expression of serum BGP, Collagen I before fracture and on the 3d, 5th, 7th,14th, 21st, 28th, 35th day after fracture (n=6/time point). Callus of fracture sides weredetermined by the TGF-β1 mRNA and protein expression on the 3d, 5th, 7th, 14th, 21st, 28th,35th day (n=6/time point) via reverse transcriptase polymerase chain reaction (RT-PCR) andenzyme linked immunosorbent assay (Elisa).Results: HXHY decoction can promote the expressions of serum BGP, Collagen I,TGF-β1 of callus of fracture sides. Serum Collagen I reaches low value on the 7th day,increases gradually and peak value on the 21st day, after 21d, falls gradually and approachesthe numerical value in gear on the 35th day after fracture separately. Two groups are inaccordant trend. After treatment of HXHY decoction, the expressions of serum Collagen Iincreased significantly on the 5th, 7th, 14th, 21st day (p＜0.01); on the 3d, 28th, 35th day, the differences of them are not significant (p＞0.05) during fracture healing. The expressions ofserum BGP is in accord with Collagen I. All callus tissue TGF-β1 mRNA expressionincreased on the 3d, 14th day at the peak, especially on the 14th day. There is an significantlydecrease on TGF-β1 mRNA expression especially on the 7th day. The expressions of TGF-β1mRNA is in accord with its protein. The expressions of TGF-β1 mRNA and its proteinincreased significantly on the 3d, 14th (p＜0.01), 21st day (p＜0.05), decreased significantly onthe 5th, 7th day in HXHY decoction group at the time compared with the normal sodium group.HXHY decoction can shorten the time of fracture healing and moulding, improve the quality ofcicatrized bone.Conclusions:1. Standardized percutaneous intramedullary nail fracture model of the tibia in ratsusing a modified technique fixed have the characters of stability, eliability, repeatability.2. HXHY decoction promoting the expressions of serum BGP, Collagen I, TGF-β1 ofcallus of fracture sides as to accelerate endochondral ossification.3. HXHY decoction can shorten the time of fracture healing and moulding, improve thequality of cicatrized bone.4. TGF-β1 is a representative gene in the process of endochondral ossification duringfracture healing.5. The expressions of TGF-β1 mRNA and its protein decreased significantly on the 5th, 7thday in HXHY decoction group at the time compared with the normal sodium group duringearly fracture healing. There might be through System Ihh (Indian hedgehog)/PTHrP(PTH-related peptide), System TGF-β/BMPs, the number of combinations between TGF-β1and its receptors.
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