【摘要】：目的:通過觀察富硒地龍對(duì)大鼠左側(cè)股骨骨折愈合過程中骨形態(tài)發(fā)生蛋白-2與血管內(nèi)皮生長(zhǎng)因子表達(dá)的影響,探討富硒地龍對(duì)骨折愈合促進(jìn)作用及其可能的作用機(jī)制。方法:選取雄性SD大鼠72只,制作左側(cè)股骨干骨折模型。模型復(fù)制成功后將其分為傷科接骨片組、富硒地龍組、生理鹽水組,每組24只。術(shù)后分別予以富硒地龍300 mg/(kg.d)-1,傷科接骨片300 mg/(kg.d)-1,生理鹽水組予以等容量生理鹽水灌胃干預(yù)。于術(shù)后2、4、6周每組各取8只大鼠對(duì)股骨大體標(biāo)本進(jìn)行觀察、骨痂體積測(cè)量、DR攝片和HE染色,血清生化檢測(cè)VEGF、免疫組化檢測(cè)BMP-2表達(dá)情況,最后統(tǒng)計(jì)分析結(jié)果。結(jié)果:1.骨折斷端影像學(xué)表現(xiàn):術(shù)后第2周各組均可見少量骨痂形成,均可見清晰的骨折線;第4周各組大部分標(biāo)本均已形成骨性骨痂,有骨痂組織連續(xù)性通過骨折斷端,骨折線模糊;第6周各組均達(dá)到骨性愈合標(biāo)準(zhǔn),骨折線消失。經(jīng)評(píng)分統(tǒng)計(jì),各期各組間比較均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。2.大體標(biāo)本觀察及骨痂測(cè)量:術(shù)后第2周各組均可見明顯骨痂形成,體積大,質(zhì)地略軟;第4周各組骨痂體積均變小、質(zhì)地變硬;第6周各組骨痂體積進(jìn)一步縮小,骨痂改建、塑形,肉眼觀察各組未見區(qū)別。測(cè)量骨痂體積,經(jīng)統(tǒng)計(jì)學(xué)處理,第4周富硒地龍組與傷科接骨片組、生理鹽水組比較有統(tǒng)計(jì)學(xué)意義(P0.05)。3.骨痂組織學(xué)觀察:第2周:各組骨折斷端骨內(nèi)、外膜下均可見骨小梁形成,骨折斷端間有纖維性組織連接,富硒地龍組與傷科接骨片組膜內(nèi)成骨量及軟骨島面積明顯大于生理鹽水組;第4周:各組骨小梁進(jìn)一步增多增粗,穿插排列,連接骨折斷端;第6周:傷科接骨片組和富硒地龍組骨折斷端間骨小梁完全轉(zhuǎn)變?yōu)榘鍖訝罟?髓腔再通。生理鹽水組骨小梁逐漸向板層狀骨過渡,部分標(biāo)本髓腔再通。統(tǒng)計(jì)分析富硒地龍組和生理鹽水組有統(tǒng)計(jì)學(xué)意義(P0.05)。4.免疫組化檢測(cè)情況:各組骨痂組織中成纖維細(xì)胞、軟骨細(xì)胞、成骨細(xì)胞胞漿中均可見BMP-2陽(yáng)性表達(dá),初期以成骨細(xì)胞、骨小梁中骨細(xì)胞及近骨痂外周幼稚軟骨細(xì)胞表達(dá)為主,后期以成骨細(xì)胞表達(dá)為主,各期傷科接骨片組與富硒地龍組胞漿著色深度明顯大于生理鹽水組。測(cè)量各組BMP-2陽(yáng)性表達(dá)MOD值在第2周表現(xiàn)最高,4周開始緩慢下降,6周下降最明顯,陽(yáng)性表達(dá)最低。經(jīng)統(tǒng)計(jì)學(xué)處理,第2周:富硒地龍組與傷科接骨片組、生理鹽水組比較均有統(tǒng)計(jì)學(xué)意義(P0.05);第4、6周;富硒地龍組與傷科接骨片組、生理鹽水組比較有統(tǒng)計(jì)學(xué)意義(P0.05)。5.血清生化學(xué)情況:生理鹽水組大鼠外周血清中VEGF表達(dá)與正常組接近;富硒地龍組各個(gè)識(shí)相點(diǎn)外周血清中VEGF表達(dá)顯著升高于生理鹽水組(P0.05),傷科接骨片組各個(gè)識(shí)相點(diǎn)外周血清中VEGF表達(dá)均高于模型組(P0.05)。結(jié)論:富硒地龍水溶液可以促進(jìn)骨痂組織中BMP-2和血清中VEGF表達(dá);這可能是治療骨折愈合的機(jī)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.
【學(xué)位授予單位】：湖北民族學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285.5

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