辛伐他

发布时间：2018-08-24 09:08

【摘要】：目的本研究通过建立大鼠Ⅰ型糖尿病下颌骨骨折模型，观察辛伐他汀与胰岛素联合用药对糖尿病大鼠下颌骨骨折愈合过程中骨痂形态、骨密度的影响，联合检测血清中碱性磷酸酶及骨钙素浓度、骨痂中骨钙素的表达情况，以探讨辛伐他汀对糖尿病下颌骨骨折是否有促进愈合作用，，为糖尿病病人选择促进骨愈合药物提供理论依据。方法选用6～8周龄，体重300±20gWistar大鼠96只，按随机原则分为4组，对照组（A组），糖尿病骨折组（B组），糖尿病骨折+胰岛素组（C组），糖尿病骨折+胰岛素+辛伐他汀组（D组），每组24只。适应性喂养2w后，开始Ⅰ型糖尿病动物模型的建立。分别给予糖尿病骨折组、糖尿病骨折+胰岛素组、糖尿病骨折+胰岛素+辛伐他汀组大鼠一次性大剂量腹腔注射链脲佐菌素（STZ）50mg/kg。三天后，测定三组大鼠的血糖浓度，血糖浓度≥16.7mmol/L视作造模成功。恢复饲养3d后，开始四组大鼠的下颌骨骨折模型建立。全麻下用打磨机金刚砂片在大鼠下颌骨体下缘咬肌嵴的远中末端处制作1mm宽、3mm长的贯穿颊舌侧的不完全骨缺损。术后，糖尿病骨折+胰岛素组及糖尿病骨折+胰岛素+辛伐他汀组给予胰岛素控制血糖。辛伐他汀配成悬浊液，给予糖尿病骨折+胰岛素+辛伐他汀组每天10mg/kg灌胃，余下各组予以等量生理盐水灌胃。分别于术后2w，4w，6w，8w每组各处死大鼠6只，抽取血清样本，使用ELISA试剂盒测得血清中ALP及BGP的浓度。取大鼠的下颌骨标本行骨密度和X线片检查，然后置于4％多聚甲醛中固定48h，再使用10％乙二胺四乙酸进行为期60d的组织脱钙。脱钙完成后，组织块石蜡包埋行HE染色组织形态学观察及BGP免疫组织化学检测。结果1除A组外，余下三组大鼠在注射链脲佐菌素72h后，测得血糖浓度均高于16.7mmol/L，并逐渐出现典型的糖尿病“三多一少”症状。骨折模型建立后，庆大霉素肌注1w，周围软组织伤口无红肿，无感染。2成模后于2w，4w，6w，8w在各组间进行X线比较：2w时各组间X线片显示大鼠骨折线密度均较低，4w，6w，8w时D组均较B组骨折线密度增高，骨折线模糊。骨密度比较：术后4w，6w，8w时D组测得骨密度高于B组（P＜0.05），有统计学意义。4w时，D组较A组骨密度低（P＜0.05），有统计学意义。8w时，D组较C组骨密度高（P＜0.05），有统计学意义。血清骨钙素浓度比较：2w、4w、6w、8w时，D组大鼠血清骨钙素浓度均高于B组（P＜0.05），有统计学意义。2w、4w时，D组血清骨钙素浓度高于C组（P＜0.05），有统计学意义。血清碱性磷酸酶浓度比较：2w，4w，6w，8w时，D组血清碱性磷酸酶浓度均高于B组（P＜0.05），有统计学意义。2w、4w时，D组高于C组（P＜0.05），有统计学意义。组织形态学观察：D组在2w，4w，6w，8w里骨小梁数量、骨小梁宽度、骨小梁长度等均优于B组。免疫组化检测：2w，4w，6w时，D组骨痂区骨钙素的表达均高于B组、C组（P＜0.05），有统计学意义。结论1通过一次性大剂量注射STZ的方法可以快速诱导建立糖尿病大鼠模型及通过大鼠下颌骨体制作1mm×3mm贯穿大鼠颊舌侧的骨缺损，可以建立下颌骨不完全性骨折实验动物模型。2动物实验证实糖尿病各组骨折愈合情况较对照组差，糖尿病可导致骨愈合延迟。3辛伐他汀可提高糖尿病大鼠骨折区BGP和ALP的表达，辛伐他汀与胰岛素联合用药可促进Ⅰ型糖尿病大鼠下颌骨骨折的愈合。
[Abstract]:Objective To investigate the effects of simvastatin combined with insulin on callus morphology and bone mineral density during mandibular fracture healing in diabetic rats by establishing a model of type I diabetic mandibular fracture in rats. Whether Ting can promote the healing of diabetic mandibular fracture provides a theoretical basis for diabetic patients to choose drugs to promote bone healing.
Methods Ninety-six Wistar rats aged 6-8 weeks and weighing 300+20 g were randomly divided into 4 groups: control group (group A), diabetic fracture group (group B), diabetic fracture + insulin group (group C), diabetic fracture + insulin + simvastatin group (group D), 24 rats in each group. After 2 weeks of adaptive feeding, the animal model of type I diabetes mellitus was established. After three days of intraperitoneal injection of streptozotocin (STZ) 50 mg/kg, the blood glucose concentrations of the three groups were determined. The concentration of blood glucose (> 16.7 mmol/L) was regarded as a successful model. The mandibular fracture models of the four groups were established after 3 days of recovery. The distal end of the masseter ridge at the lower margin of the mandible of rats was made with grinding machine diamond tablets under general anesthesia to produce incomplete bone defect with a width of 1 mm and a length of 3 mm through the cheek and tongue. After operation, the diabetic fracture + insulin group and the diabetic fracture + insulin + Simvastatin group were given insulin to control blood sugar. Six rats in each group were sacrificed at 2w, 4w, 6W and 8W after operation. Serum samples were taken and the concentrations of ALP and BGP in serum were measured by ELISA kit. Bone mineral density (BMD) and X-ray films were taken from the mandibular bone specimens of the rats and placed in 4% of the rats. Polyformaldehyde was immobilized for 48 hours and then decalcified with 10% ethylenediaminetetraacetic acid for 60 days.
Results 1 Except group A, the blood glucose concentration of the other three groups was higher than 16.7 mmol/L 72 hours after injection of streptozotocin, and the typical symptoms of diabetes mellitus gradually appeared. After the establishment of the fracture model, gentamicin was injected intramuscularly for 1 week, and the wounds around the soft tissue were not inflamed and inflamed. The bone mineral density of group D was higher than that of group B at 4w, 6W and 8W after operation (P < 0.05). The bone mineral density of group D was higher than that of group B at 4w, 6W and 8W after operation (P < 0.05). The bone mineral density of group D was lower than that of group A at 4W (P < 0.05). The serum levels of osteocalcin in group D were higher than those in group B at 2w, 4w, 6W and 8W (P < 0.05). The serum levels of osteocalcin in group D were higher than those in group C at 2w, 4w, 6W and 8W (P < 0.05). The concentration of phosphatase in group D was higher than that in group B (P Group P (0.05) was statistically significant.
Conclusion 1 The diabetic rat model can be established by injecting STZ in a large dose at once and the 1 mm 65 Urinary disease can lead to delayed bone healing. 3 Simvastatin can increase the expression of BGP and ALP in the fracture area of diabetic rats. Simvastatin combined with insulin can promote the healing of mandibular fracture in type I diabetic rats.
【学位授予单位】：河北联合大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.1;R782.4

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