不同的补肾方法对大鼠骨折愈合过程中Osx、TGF-β1表达影响的实验研究
发布时间：2018-07-29 05:37
【摘要】：目的：建立大鼠左侧胫骨中段骨折的动物模型，观察不同的补肾方法对大鼠骨折愈合过程中Osx、TGF-β1表达的影响，为其能促进成骨细胞分化，促进骨折愈合提供一定的理论依据。 方法：选用250±20g，SPF级wistar大鼠96只，随机分为4组，每组24只，分别为A蒸馏水组、B补肾壮筋汤组、C左归丸组、D右归丸组。采用三点弯曲法造成左胫骨中段闭合性骨折的动物模型，术后用自制小夹板（长2cm、宽0.25cm三合板制成）固定。造模6小时后给予药物干预，分别于术后1、2、4周每组8只大鼠经腹腔麻醉后行X线检查、骨折处骨痂定量的比较以观察不同的补肾方药对骨折愈合的疗效差异，然后处死动物取出骨质标本，并放置于4%多聚甲醛中；制作切片，采用免疫组化方法处理，观察各组大鼠骨痂的Osx、TGF-β1因子表达情况，并用ImagePro plus6.图像分析软件对骨痂标本阳性表达细胞的平均光密度值（MOD）进行测定。对实验数据采用spss17.0软件进行统计分析。 结果： X线片检查，B、C、D用药组与A蒸馏水组比较，术后第1周骨折断端的X线影像学无明显差异，术后第2、4周B、C、D用药组骨折愈合快于A蒸馏水组，B组骨折愈合快于C、D组，C组骨折愈合快于D组。各时段骨折断端骨痂定量比较，术后第1周各组无明显统计学意义，术后第2、4周B组骨折断端骨痂含量显著高于A组(P＜0.01)，C、D组骨折断端骨痂含量明显高于A组(P＜0.05)，B组骨折断端骨痂含量明显高于C、D组(P＜0.05)，C组骨折断端骨痂含量明显高于D组(P＜0.05)。A、B、C、D组骨痂标本免疫组织化学结果提示：各组Osx、TGF-β1表达水平从术后第1周到第2周逐渐增强，术后第4周呈现下降趋势。术后1、2、4周B、C、D组的Osx、TGF-β1的阳性细胞数的平均光密度值（MOD）显著高于A组(P＜0.05)，B组Osx、TGF-β1的阳性细胞数的平均光密度值（MOD）明显高于C、D组(P＜0.05)，C组Osx、TGF-β1的阳性细胞数的平均光密度值（MOD）明显高于D组(P＜0.05)，，均在2周时阳性表达达到高峰。 结论：补肾壮筋汤、左归丸、右归丸均有促进Osx、TGF-β1表达作用，补肾壮筋汤较左归丸、右归丸促进骨折愈合过程中Osx、TGF-β1因子表达水平显著。补肾壮筋汤比左归丸、右归丸更能加速骨折局部骨痂的形成，促进骨折愈合。
[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.
【学位授予单位】：甘肃中医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5;R274.9
本文编号：2151690
[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.
【学位授予单位】：甘肃中医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5;R274.9
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