PTH对高脂STZ诱导并卵巢切除大鼠闭合骨折愈合的影响

发布时间：2018-05-29 05:24

本文选题：高脂STZ + 卵巢切除术　；参考：《华北理工大学》2016年硕士论文

【摘要】：目的研究甲状旁腺激素(Parathyroid hormone PTH)对高脂STZ诱导并卵巢切除大鼠骨折愈合的影响。方法将健康的50只2月龄雌性SD大鼠进行适应(环境和温度以及湿度)饲养7天后,随机进行对大鼠分为5组,进行实验研究。骨折+假手术组(F+Sham组)10只,骨折+双侧卵巢切除组(F+OVX组)10只,骨折+假手术+高脂STZ诱导(F+Sham+STZ组)10只,骨折+高脂STZ诱导+双侧卵巢切除(F+STZ+OVX组)10只,骨折+双侧卵巢切除+高脂STZ诱导+甲状旁腺激素(F+OVX+STZ+PTH组)10只,各组大鼠均在骨折造模立刻行X线摄像评估右侧股骨骨折情况。F+Sham和F+OVX组在三周腹腔注射柠檬酸缓冲液1ml/kg,而其他三组在3w腹腔注射STZ 35mg/kg。F+OVX+STZ+PTH组的闭合骨折造模手术后给予皮下注射PTH剂量为50ug/kg,每周5次,F+STZ+OVX组的闭合骨折造模手术后皮下注射等量的生理盐水,每周5次。给药三周后,处死前各组大鼠行右侧股骨X线摄像,对骨折断端愈合进行放射学评分。应用Image J软件分析骨痂面积。处死动物后留取右侧股骨行骨组织切片番红-o-固绿染色,免疫组化分析骨折断端VEGF表达情况。结果1 X线检测结果显示F+Sham组骨折断端边缘完全消失,骨膜反应密度近似骨影,骨痂填满缺损,与骨皮质密度基本相同并相互连接,F+STZ和F+OVX及F+STZ+OVX组可见骨骼组织骨折端骨痂组织细胞形成,但骨折组织断端无连接,F+OVX+STZ+PTH组可见骨骼组织骨折端骨痂形成及骨痂组织细胞数量增多,骨折组织断端可见小梁骨桥接形成骨连接,骨连接密度高。与F+Sham组相比,其余组骨折X线评分显著低(P0.05),F+STZ+OVX与F+OVX+STZ+PTH组相比骨折X线评分显著低(P0.05)。2骨骼组织骨折愈合骨痂的面积评估显示F+Sham与F+STZ、F+STZ+OVX相比骨痂面积值高(P0.05),F+OVX与F+STZ+OVX相比骨骼组织骨痂面积低(P0.05),F+STZ+OVX与F+OVX+STZ+PTH组相比骨痂面积值升高(P0.05)。3免疫组织化结果显示F+Sham组骨骼组织股骨干骨折断端可见少量的VEGF的阳性表达,与F+Sham组相比F+STZ骨折断端骨组织细胞中VEGF的阳性表达下降,与F+Sham组相比F+OVX组可见VEGF的阳性表达也下降,与F+STZ组相比,F+STZ+OVX组可见VEGF的阳性表达也下降,与F+STZ+OVX组相比,F+OVX+STZ+PTH组骨折断端VEGF的阳性表达升高。结论1高脂STZ诱导并卵巢切除大鼠导致闭合性骨折断端延期愈合。2 PTH治疗可以促进高脂STZ诱导并卵巢切除大鼠闭合性骨折断端愈合,这可能与其上调骨痂中VEGF的表达水平有关。
[Abstract]:Objective to study the effect of parathyroid hormone (PTH) on fracture healing induced by hyperlipidemic STZ in ovariectomized rats. Methods 50 healthy 2-month-old female SD rats were fed with adaptation (environment, temperature and humidity) for 7 days, and were randomly divided into 5 groups. There were 10 rats in the sham operation group, 10 in the bilateral ovariectomy group, 10 in the F OVX group, 10 in the high fat STZ induced F Sham STZ group, and 10 in the bilateral ovariectomized STZ OVX group induced by high fat STZ. 10 rats were treated with bilateral ovariectomized with hyperlipidemic STZ induced parathyroid hormone (PTH) F OVX STZ PTH group (n = 10). All rats in each group were immediately treated with X-ray imaging to evaluate the fracture of the right femur. F Sham and F OVX groups were intraperitoneally injected with citric acid buffer solution of 1 ml / kg for three weeks, while the other three groups received intraperitoneal injection of closed fracture in STZ 35mg/kg.F OVX STZ PTH group for 3 weeks. PTH was subcutaneously injected with 50ugr / kg after model operation, and the same amount of normal saline was injected subcutaneously after closed fracture modeling operation in F STZ OVX group 5 times a week. Five times a week. After three weeks of administration, the rats in each group were given right femur X-ray before death, and the fracture healing was scored radiologically. The callus area was analyzed by Image J software. After the animals were killed, the right femur was taken to be stained with Fenghong-o-solid green staining and the expression of VEGF was analyzed by immunohistochemistry. Results 1the results of X-ray examination showed that the edge of fracture end disappeared completely, the periosteal reaction density was similar to bone shadow, and callus was filled with defect in F Sham group. Bone callus cells formed at the end of bone fracture were observed in F STZ, F OVX and F STZ OVX groups with the same density of bone cortex. In F OVX STZ PTH group, the formation of bone callus and the number of callus cells were increased. Trabecular bone bridging was seen in the broken end of fracture tissue, and the density of bone connection was high. Compared with F Sham group, Fracture X-ray score of the other groups was significantly lower than that of the F OVX STZ PTH group, the X-ray score of the fracture was significantly lower than that of the F OVX STZ PTH group. The evaluation of the area of bone callus healing of bone tissue showed that F Sham and F STZ F STZ OVX had higher callus area than that of F Sham and F STZ F STZ OVX (P 0.05 / F OVX and F STZ). Compared with the F OVX STZ PTH group, the bone callus area of OVX group was lower than that of the bone tissue callus area. The immunohistochemical results showed that a small amount of VEGF positive expression was found in the fracture end of the femoral shaft fracture in the F Sham group, compared with the value of the bone callus area in the F OVX STZ PTH group. Compared with F Sham group, the positive expression of VEGF in bone tissue of F STZ fracture was decreased, the positive expression of VEGF in F OVX group was lower than that in F Sham group, and the positive expression of VEGF in F STZ OVX group was lower than that in F STZ group. The positive expression of VEGF in fracture end of F OVX STZ PTH group was higher than that in F STZ OVX group. Conclusion (1) delayed healing of closed fracture induced by hyperlipidemic STZ and ovariectomized rats could promote the healing of closed fracture in hyperlipidemic STZ and ovariectomized rats, which may be related to the up-regulation of VEGF expression in callus.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R58;R683

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