發(fā)布時間：2018-03-12 12:02

  本文選題：二膦酸鹽　切入點(diǎn)：糖尿病　出處：《河北聯(lián)合大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的本實(shí)驗通過建立大鼠Ⅰ型糖尿病下頜骨骨折模型，觀察阿侖膦酸鈉與胰島素聯(lián)合應(yīng)用于Ⅰ型糖尿病大鼠下頜骨骨折愈合的過程，探討阿侖膦酸鈉在糖尿病干擾下對大鼠下頜骨骨折愈合的影響，為糖尿病頜骨骨折篩選臨床用藥提供依據(jù)及動物實(shí)驗數(shù)據(jù)。 方法選用6～8周齡，體重300±20gWistar大鼠96只，隨機(jī)分為4組，空白對照組（A組），糖尿病骨折組（B組），糖尿病骨折+胰島素組（C組），糖尿病骨折+胰島素+阿侖膦酸鈉組（D組），每組24只。適應(yīng)性喂養(yǎng)兩周后，應(yīng)用鏈脲佐菌素（STZ）大劑量一次性腹腔注射，建立I型糖尿病動物模型。三天后，，測定隨機(jī)血糖濃度，根據(jù)相關(guān)文獻(xiàn)，血糖濃度16.7mmol/L大鼠，視為造模成功�；謴�(fù)飼養(yǎng)3d后，10%水合氯醛全身麻醉下行大鼠左側(cè)下頜骨骨折模型造模術(shù)。術(shù)后，除空白對照組及糖尿病骨折組外，其余各組給予胰島素控糖。術(shù)后1d起糖尿病骨折+胰島素+阿侖膦酸鈉組給予35mg/kg/w阿侖膦酸鈉灌胃，其余各組給予等量的生理鹽水灌胃。術(shù)后在2w，4w，6w，8w時每組各處死大鼠6只，取材，應(yīng)用ELISA方法檢測血清中OPG及TRACP-5b含量，取出下頜骨行骨密度檢測和X線片檢查后，放入4％多聚甲醛固定48h，再用10％乙二胺四乙酸（EDTA）將骨組織脫鈣60d。脫鈣完成后，組織塊石蠟包埋行HE染色組織形態(tài)學(xué)觀察，OPG免疫組織化學(xué)檢測及TRACP-5b特異性染色。 結(jié)果1糖尿病造模成功后，大鼠出現(xiàn)典型的糖尿病“三多一少”癥狀；骨折模型建立后，傷口周圍軟組織紅腫，無感染癥狀，實(shí)驗過程中糖尿病大鼠的血糖水平較穩(wěn)定，建立糖尿病大鼠骨折模型成功。2造模成功后，2w，4w，6w，8w各組間進(jìn)行X線比較：2w時各組間X線片顯示大鼠骨折線密度均較低；4w，6w，8w時空白對照組及糖尿病骨折+胰島素+阿侖膦酸鈉組骨折區(qū)域密度相對較高，骨折線模糊。骨密度比較：術(shù)后2w，4w，糖尿病骨折+胰島素+阿侖膦酸鈉組骨密度明顯高于糖尿病骨折組及糖尿病骨折+胰島素組，有統(tǒng)計學(xué)意義。組織形態(tài)學(xué)觀察：糖尿病骨折+胰島素+阿侖膦酸鈉組在骨折2w，4w，6w時破骨細(xì)胞數(shù)目較少、骨小梁數(shù)量、骨小梁寬度、骨小梁長度等優(yōu)于糖尿病骨折組。免疫組化檢測：抗酒石酸酸性磷酸酶-5b在骨痂區(qū)2w，4w，6w時糖尿病骨折+胰島素+阿侖膦酸鈉組表達(dá)低于糖尿病骨折+胰島素組及糖尿病骨折組有統(tǒng)計學(xué)意義（P0.05）。骨保護(hù)素：在骨痂區(qū)各組間均有表達(dá)，2w，4w，6w時糖尿病骨折+胰島素+阿侖膦酸鈉組表達(dá)均高于糖尿病骨折+胰島素組及糖尿病骨折組，有統(tǒng)計學(xué)意義（P0.05），8w雖有表達(dá)，但其數(shù)值無統(tǒng)計學(xué)意義（P0.05）。 結(jié)論1一次性大劑量腹腔注射STZ以及外科手術(shù)方法的應(yīng)用可成功建立大鼠I型糖尿病下頜骨骨折模型。2糖尿病大鼠骨折愈合質(zhì)量差，比較容易出現(xiàn)愈合延遲甚至不愈合。3阿侖膦酸鈉聯(lián)合胰島素在糖尿病大鼠下頜骨骨折模型中可起到促進(jìn)骨折愈合的作用，其可能機(jī)制為阿侖膦酸鈉促進(jìn)OPG的表達(dá)及抑制TRACP-5b的釋放，抑制破骨細(xì)胞在糖尿病骨折愈合區(qū)域的過度增殖從而促進(jìn)骨愈合。
[Abstract]:Objective to establish a type I diabetic rat mandibular fracture model through the experiment, the process of observation of the combined application of alendronate and insulin on mandibular fracture healing in diabetic rats, to investigate the alendronate effects on fracture healing in diabetic rat mandibular fracture disturbance, provide the basis for screening clinical and animal experiments the data for diabetic jaws.
Methods 6~8 week old, weight 300 + 96 20gWistar rats were randomly divided into 4 groups, control group (group A), diabetic group (group B), fracture fracture and diabetes + insulin group (C group), diabetic fracture + insulin + alendronate group (D group), each group 24. To two weeks after feeding, streptozotocin (STZ) intraperitoneal injection of high dose, the establishment of animal models of type I diabetes. Three days later, the determination of random blood glucose levels, according to the related literature, the blood glucose concentration in 16.7mmol/L rats, as a successful model. The recovery process after 3D, 10% chloral hydrate aldehyde under general anesthesia in rats model of left mandible fracture model. After the operation, in addition to the control group and diabetic fracture group, the rest groups were given insulin glucose control. Postoperative 1D diabetes fracture + insulin + alendronate group were given alendronate 35mg/kg/w intragastric administration of saline, other groups were given the amount of gastric lavage After operation in 2W, 4W, 6W, 8W in each group and 6 rats were detected by ELISA OPG in serum and the content of TRACP-5b, remove the mandible bone density testing and X-ray examination, fixed in 4% paraformaldehyde and 10% 48h EDTA (EDTA) will decalcified bone tissue 60d. after the completion of decalcified tissue, paraffin embedded for HE staining and histological observation, OPG immunohistochemistry and TRACP-5b staining.
Results 1 rats, rats showed typical diabetic symptoms; fracture model, soft tissue swelling around the wound, no symptoms of infection, diabetic rats during the experiment the blood glucose level is relatively stable, the establishment of diabetic rat fracture model of.2 after successful modeling, 2W, 4W. 6W, X-ray comparison: 8W group 2W group rats X-ray showed the fracture line density were low; 4W, 6W, 8W of blank control group and DM + fracture insulin + alendronate group fracture area density is relatively high, the fracture lines blurred. Bone mineral density: 2W 4W, diabetes, fracture + insulin + alendronate group bone density was significantly higher than that of diabetic group and diabetic fracture fracture + insulin group, was statistically significant. Histological observation: diabetes fracture + insulin + alendronate group in 2W 4W, 6W fracture, osteoclast The cell number and the number of trabecular bone, trabecular thickness, trabecular bone fracture length is better than that of diabetic group. Immunohistochemical detection: tartrate resistant acid phosphatase -5b in callus area 2W, 4W, 6W + + insulin diabetes fracture in the alendronate group was lower than that of diabetes fracture + insulin group diabetes and fracture group was statistically significant (P0.05). Osteoprotegerin expression in each group, the callus area between 2W, 4W, 6W + + insulin diabetes fracture in the alendronate group expression were higher in diabetic group and diabetic insulin + fracture fracture group, there was statistical significance (P0.05), 8W is the expression, but no statistically significant values (P0.05).
Conclusion the application of 1 single dose intraperitoneal injection of STZ and surgical methods can successfully establish rat mandibular fracture type I diabetes model of.2 diabetic rats poor quality of healing fractures, more prone to delayed healing or even nonunion.3 alendronate combined with insulin in the rat mandibular fracture diabetes can promote fracture healing in the model, the possible mechanism for the expression of alendronate promote OPG and inhibiting the release of TRACP-5b, and inhibit the proliferation of osteoclasts in bone fracture healing area of diabetes can promote bone healing.

【學(xué)位授予單位】：河北聯(lián)合大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R782.4;R587.1

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