發(fā)布時間：2018-07-29 05:37

【摘要】：目的：建立大鼠左側脛骨中段骨折的動物模型，觀察不同的補腎方法對大鼠骨折愈合過程中Osx、TGF-β1表達的影響，為其能促進成骨細胞分化，促進骨折愈合提供一定的理論依據(jù)。 方法：選用250±20g，SPF級wistar大鼠96只，隨機分為4組，每組24只，分別為A蒸餾水組、B補腎壯筋湯組、C左歸丸組、D右歸丸組。采用三點彎曲法造成左脛骨中段閉合性骨折的動物模型，術后用自制小夾板（長2cm、寬0.25cm三合板制成）固定。造模6小時后給予藥物干預，分別于術后1、2、4周每組8只大鼠經腹腔麻醉后行X線檢查、骨折處骨痂定量的比較以觀察不同的補腎方藥對骨折愈合的療效差異，然后處死動物取出骨質標本，并放置于4%多聚甲醛中；制作切片，采用免疫組化方法處理，觀察各組大鼠骨痂的Osx、TGF-β1因子表達情況，并用ImagePro plus6.圖像分析軟件對骨痂標本陽性表達細胞的平均光密度值（MOD）進行測定。對實驗數(shù)據(jù)采用spss17.0軟件進行統(tǒng)計分析。 結果： X線片檢查，B、C、D用藥組與A蒸餾水組比較，術后第1周骨折斷端的X線影像學無明顯差異，術后第2、4周B、C、D用藥組骨折愈合快于A蒸餾水組，B組骨折愈合快于C、D組，C組骨折愈合快于D組。各時段骨折斷端骨痂定量比較，術后第1周各組無明顯統(tǒng)計學意義，術后第2、4周B組骨折斷端骨痂含量顯著高于A組(P＜0.01)，C、D組骨折斷端骨痂含量明顯高于A組(P＜0.05)，B組骨折斷端骨痂含量明顯高于C、D組(P＜0.05)，C組骨折斷端骨痂含量明顯高于D組(P＜0.05)。A、B、C、D組骨痂標本免疫組織化學結果提示：各組Osx、TGF-β1表達水平從術后第1周到第2周逐漸增強，術后第4周呈現(xiàn)下降趨勢。術后1、2、4周B、C、D組的Osx、TGF-β1的陽性細胞數(shù)的平均光密度值（MOD）顯著高于A組(P＜0.05)，B組Osx、TGF-β1的陽性細胞數(shù)的平均光密度值（MOD）明顯高于C、D組(P＜0.05)，C組Osx、TGF-β1的陽性細胞數(shù)的平均光密度值（MOD）明顯高于D組(P＜0.05)，，均在2周時陽性表達達到高峰。 結論：補腎壯筋湯、左歸丸、右歸丸均有促進Osx、TGF-β1表達作用，補腎壯筋湯較左歸丸、右歸丸促進骨折愈合過程中Osx、TGF-β1因子表達水平顯著。補腎壯筋湯比左歸丸、右歸丸更能加速骨折局部骨痂的形成，促進骨折愈合。
[Abstract]:Objective: to establish an animal model of left middle tibia fracture in rats and observe the effect of different methods of tonifying kidney on the expression of Osxan TGF- 尾 1 during fracture healing in rats, so as to provide a theoretical basis for promoting osteoblast differentiation and fracture healing. Methods: 96 wistar rats were randomly divided into 4 groups, 24 rats in each group. The rats were divided into A distilled water group and B Bushen Zhuangjin decoction group. The animal model of closed fracture of left middle tibia was established by three point bending method. After the operation, small splint (2 cm in length and wide 0.25cm plate) was used to fix the fracture. After 6 hours of model making, 8 rats in each group were given X-ray examination after intraperitoneal anesthesia at 1, 2 and 4 weeks after operation. The bone callus at fracture site was compared quantitatively to observe the difference of curative effect of different prescription of tonifying kidney on fracture healing. Then, the bone specimens were taken out and placed in 4% paraformaldehyde. Sections were made and treated with immunohistochemical method. The expression of OsxTGF- 尾 1 in the callus of rats was observed and ImagePro plus6 was used to observe the expression of OsxTGF- 尾 1 factor in the callus of rats. The average optical density (MOD) of callus positive cells was measured by image analysis software. The experimental data were analyzed by spss17.0 software. Results: compared with A distilled water group, there was no significant difference in X-ray imaging of fracture at the first week after operation. The fracture healing of group B was faster than that of group B and group C was faster than that of group D at 2 weeks after operation. There was no statistical significance in each group at the first week after operation. The callus content at the end of fracture in group B was significantly higher than that in group A (P < 0.01) at the 2nd week after operation. The content of callus at the end of fracture in group C was significantly higher than that in group A (P < 0.05). The content of callus at the end of fracture in group C was significantly higher than that in group C (P < 0.05). The results of immunohistochemistry showed that the expression level of Osxfen TGF- 尾 1 increased gradually from the first week to the second week after operation in group D (P < 0.05), and the expression of Osxtgf- 尾 1 in group D was significantly higher than that in group D (P < 0.05). There was a downward trend at the 4th week after operation. The average optical density (MOD) of the number of OsxtGF- 尾 1 positive cells in group B was significantly higher than that in group A (P < 0. 05). The average optical density of TGF- 尾 1 cells in group B was significantly higher than that in group C (P < 0. 05), and the average optical density of TGF- 尾 1 in group C was significantly higher than that in group C (P < 0. 05). The value of (MOD) was significantly higher than that of group D (P < 0. 05), and the positive expression reached the peak at 2 weeks. Conclusion: Bushen Zhuangjin decoction, Zuogui Pill and Yougui Pill can all promote the expression of OsxTGF- 尾 1, and Bushen Zhuangjin decoction is more effective than Zuo Gui Pill, and Yougui Pill can promote the expression level of OsxTGF- 尾 1 factor in fracture healing process. Bushen Zhuangjin decoction can accelerate the formation of local callus and promote fracture healing more than Zuo Gui Pill and Yougui Pill.
【學位授予單位】：甘肅中醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R285.5;R274.9

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