富硒地龙对大鼠骨折BMP-2、VEGF表达的影响
发布时间：2019-05-17 14:52
【摘要】：目的:通过观察富硒地龙对大鼠左侧股骨骨折愈合过程中骨形态发生蛋白-2与血管内皮生长因子表达的影响,探讨富硒地龙对骨折愈合促进作用及其可能的作用机制。方法:选取雄性SD大鼠72只,制作左侧股骨干骨折模型。模型复制成功后将其分为伤科接骨片组、富硒地龙组、生理盐水组,每组24只。术后分别予以富硒地龙300 mg/(kg.d)-1,伤科接骨片300 mg/(kg.d)-1,生理盐水组予以等容量生理盐水灌胃干预。于术后2、4、6周每组各取8只大鼠对股骨大体标本进行观察、骨痂体积测量、DR摄片和HE染色,血清生化检测VEGF、免疫组化检测BMP-2表达情况,最后统计分析结果。结果:1.骨折断端影像学表现:术后第2周各组均可见少量骨痂形成,均可见清晰的骨折线;第4周各组大部分标本均已形成骨性骨痂,有骨痂组织连续性通过骨折断端,骨折线模糊;第6周各组均达到骨性愈合标准,骨折线消失。经评分统计,各期各组间比较均无统计学意义(P0.05)。2.大体标本观察及骨痂测量:术后第2周各组均可见明显骨痂形成,体积大,质地略软;第4周各组骨痂体积均变小、质地变硬;第6周各组骨痂体积进一步缩小,骨痂改建、塑形,肉眼观察各组未见区别。测量骨痂体积,经统计学处理,第4周富硒地龙组与伤科接骨片组、生理盐水组比较有统计学意义(P0.05)。3.骨痂组织学观察:第2周:各组骨折断端骨内、外膜下均可见骨小梁形成,骨折断端间有纤维性组织连接,富硒地龙组与伤科接骨片组膜内成骨量及软骨岛面积明显大于生理盐水组;第4周:各组骨小梁进一步增多增粗,穿插排列,连接骨折断端;第6周:伤科接骨片组和富硒地龙组骨折断端间骨小梁完全转变为板层状骨,髓腔再通。生理盐水组骨小梁逐渐向板层状骨过渡,部分标本髓腔再通。统计分析富硒地龙组和生理盐水组有统计学意义(P0.05)。4.免疫组化检测情况:各组骨痂组织中成纤维细胞、软骨细胞、成骨细胞胞浆中均可见BMP-2阳性表达,初期以成骨细胞、骨小梁中骨细胞及近骨痂外周幼稚软骨细胞表达为主,后期以成骨细胞表达为主,各期伤科接骨片组与富硒地龙组胞浆着色深度明显大于生理盐水组。测量各组BMP-2阳性表达MOD值在第2周表现最高,4周开始缓慢下降,6周下降最明显,阳性表达最低。经统计学处理,第2周:富硒地龙组与伤科接骨片组、生理盐水组比较均有统计学意义(P0.05);第4、6周;富硒地龙组与伤科接骨片组、生理盐水组比较有统计学意义(P0.05)。5.血清生化学情况:生理盐水组大鼠外周血清中VEGF表达与正常组接近;富硒地龙组各个识相点外周血清中VEGF表达显著升高于生理盐水组(P0.05),伤科接骨片组各个识相点外周血清中VEGF表达均高于模型组(P0.05)。结论:富硒地龙水溶液可以促进骨痂组织中BMP-2和血清中VEGF表达;这可能是治疗骨折愈合的机制。
[Abstract]:Aim: to observe the effect of selenium-enriched Dilong on the expression of bone morphogenetic protein-2 and vascular endothelial growth factor (VEGF) in the process of left femoral fracture healing in rats, and to explore the promoting effect of selenium-enriched Dilong on fracture healing and its possible mechanism. Methods: 72 male SD rats were selected to make the model of left thigh shaft fracture. After the successful replication of the model, the model was divided into three groups: injured Jiegu tablet group, selenium-enriched Dragon Group group and saline group with 24 rats in each group. After operation, selenium-enriched Dilong 300 mg/ (kg.d)-1, Shangke Jiegu tablet 300 mg/ (kg.d)-1, saline group were given the same volume of saline intragastrically. At 2, 4 and 6 weeks after operation, 8 rats in each group were taken to observe the gross specimens of femur, the volume of bone eschar was measured, DR film and HE staining, serum biochemical detection of BMP-2 expression was detected by VEGF, and the results were statistically analyzed. Result: 1. Imaging findings of fracture end: at the 2nd week after operation, a small amount of callus could be seen in each group, and clear broken bone lines could be seen in all groups, and in the 4th week, most of the specimens in each group had formed bone eschar, and the fracture line was blurred with the continuity of callus tissue passing through the broken end of fracture. At the 6th week, the bone healing standard was reached in all groups, and the fracture line disappeared. According to the score statistics, there was no significant difference among the groups in each period (P 0.05). 2. Gross specimen observation and eschar measurement: at the 2nd week after operation, the callus formation was obvious in each group, the volume was large and the texture was slightly soft, and the volume and texture of each group became smaller and harder at the 4th week. At the 6th week, the volume of eschar was further reduced, the eschar was rebuilt and molded, and there was no difference between the two groups. The volume of eschar was measured. At the 4th week, there was significant difference between the selenium-enriched Dragon Group group and the injured bone tablet group and saline group (P 0.05). Histologic observation of eschar: at the 2nd week, the formation of bone trabeculae could be seen in the broken bone and under the outer membrane of each group, and there was a fibrous tissue connection between the broken ends of the fracture. The amount of bone formation in the membrane and the area of cartilage island in the selenium-enriched Dragon Group group and the injured group were significantly larger than those in the saline group. Week 4: the bone trabeculae of each group were further thickened, inserted and arranged, and connected to the broken end of the fracture. At the 6th week, the bone trabeculae between the fracture ends of the injured bone group and the selenium-enriched Dragon Group fracture were completely transformed into lamellar bone, and the pulp cavity was recanalized. In saline group, the bone trabeculae gradually transitioned to lamellar bone, and some of the specimens were recanalized. Statistical analysis of selenium-enriched Dragon Group and saline group was statistically significant (P 0.05). 4. The expression of BMP-2 was positive in fibroblasts, chondrocytes and osteoblasts in eschar tissues of each group, and mainly in osteoblasts, osteocytes in trabecular meshwork and immature cartilage cells near scab in the early stage of the expression of bFGF in osteoblasts, osteoblasts and osteoblasts in the cytoplasm of osteoblasts, osteoblasts and osteoblasts in the cytoplasm of osteoblasts, osteoblasts and osteoblasts in each group. Osteoblasts were mainly expressed in the later stage, and the depth of cytoplasmic staining of selenium-enriched Dragon Group in Jiegu tablet group and selenium-enriched group was significantly higher than that in saline group. The MOD value of BMP-2 positive expression in each group was the highest at the 2nd week, began to decrease slowly at the 4th week, decreased most significantly at the 6th week, and the positive expression was the lowest. After statistical treatment, the second week: there was significant difference between selenium-enriched Dragon Group group and Shangke Jiegu tablet group and saline group (P 0.05), 4 and 6 weeks, and the difference between the two groups was statistically significant (P 0.05), and there was significant difference between the two groups (P 0.05). There was significant difference between selenium-enriched Dragon Group and Shangke Jiegu tablet group and saline group (P 0.05). The expression of VEGF in peripheral serum of saline group was similar to that of normal group. The expression of VEGF in peripheral serum of selenium-enriched Dragon Group was significantly higher than that of saline group (P 0.05), and the expression of VEGF in peripheral serum of Shangke Jiegu tablet group was higher than that of model group (P 0.05). Conclusion: selenium-enriched Dilong solution can promote the expression of BMP-2 in eschar and VEGF in serum, which may be the mechanism of fracture healing.
【学位授予单位】：湖北民族学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5
本文编号：2479183
[Abstract]:Aim: to observe the effect of selenium-enriched Dilong on the expression of bone morphogenetic protein-2 and vascular endothelial growth factor (VEGF) in the process of left femoral fracture healing in rats, and to explore the promoting effect of selenium-enriched Dilong on fracture healing and its possible mechanism. Methods: 72 male SD rats were selected to make the model of left thigh shaft fracture. After the successful replication of the model, the model was divided into three groups: injured Jiegu tablet group, selenium-enriched Dragon Group group and saline group with 24 rats in each group. After operation, selenium-enriched Dilong 300 mg/ (kg.d)-1, Shangke Jiegu tablet 300 mg/ (kg.d)-1, saline group were given the same volume of saline intragastrically. At 2, 4 and 6 weeks after operation, 8 rats in each group were taken to observe the gross specimens of femur, the volume of bone eschar was measured, DR film and HE staining, serum biochemical detection of BMP-2 expression was detected by VEGF, and the results were statistically analyzed. Result: 1. Imaging findings of fracture end: at the 2nd week after operation, a small amount of callus could be seen in each group, and clear broken bone lines could be seen in all groups, and in the 4th week, most of the specimens in each group had formed bone eschar, and the fracture line was blurred with the continuity of callus tissue passing through the broken end of fracture. At the 6th week, the bone healing standard was reached in all groups, and the fracture line disappeared. According to the score statistics, there was no significant difference among the groups in each period (P 0.05). 2. Gross specimen observation and eschar measurement: at the 2nd week after operation, the callus formation was obvious in each group, the volume was large and the texture was slightly soft, and the volume and texture of each group became smaller and harder at the 4th week. At the 6th week, the volume of eschar was further reduced, the eschar was rebuilt and molded, and there was no difference between the two groups. The volume of eschar was measured. At the 4th week, there was significant difference between the selenium-enriched Dragon Group group and the injured bone tablet group and saline group (P 0.05). Histologic observation of eschar: at the 2nd week, the formation of bone trabeculae could be seen in the broken bone and under the outer membrane of each group, and there was a fibrous tissue connection between the broken ends of the fracture. The amount of bone formation in the membrane and the area of cartilage island in the selenium-enriched Dragon Group group and the injured group were significantly larger than those in the saline group. Week 4: the bone trabeculae of each group were further thickened, inserted and arranged, and connected to the broken end of the fracture. At the 6th week, the bone trabeculae between the fracture ends of the injured bone group and the selenium-enriched Dragon Group fracture were completely transformed into lamellar bone, and the pulp cavity was recanalized. In saline group, the bone trabeculae gradually transitioned to lamellar bone, and some of the specimens were recanalized. Statistical analysis of selenium-enriched Dragon Group and saline group was statistically significant (P 0.05). 4. The expression of BMP-2 was positive in fibroblasts, chondrocytes and osteoblasts in eschar tissues of each group, and mainly in osteoblasts, osteocytes in trabecular meshwork and immature cartilage cells near scab in the early stage of the expression of bFGF in osteoblasts, osteoblasts and osteoblasts in the cytoplasm of osteoblasts, osteoblasts and osteoblasts in the cytoplasm of osteoblasts, osteoblasts and osteoblasts in each group. Osteoblasts were mainly expressed in the later stage, and the depth of cytoplasmic staining of selenium-enriched Dragon Group in Jiegu tablet group and selenium-enriched group was significantly higher than that in saline group. The MOD value of BMP-2 positive expression in each group was the highest at the 2nd week, began to decrease slowly at the 4th week, decreased most significantly at the 6th week, and the positive expression was the lowest. After statistical treatment, the second week: there was significant difference between selenium-enriched Dragon Group group and Shangke Jiegu tablet group and saline group (P 0.05), 4 and 6 weeks, and the difference between the two groups was statistically significant (P 0.05), and there was significant difference between the two groups (P 0.05). There was significant difference between selenium-enriched Dragon Group and Shangke Jiegu tablet group and saline group (P 0.05). The expression of VEGF in peripheral serum of saline group was similar to that of normal group. The expression of VEGF in peripheral serum of selenium-enriched Dragon Group was significantly higher than that of saline group (P 0.05), and the expression of VEGF in peripheral serum of Shangke Jiegu tablet group was higher than that of model group (P 0.05). Conclusion: selenium-enriched Dilong solution can promote the expression of BMP-2 in eschar and VEGF in serum, which may be the mechanism of fracture healing.
【学位授予单位】：湖北民族学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5
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