發(fā)布時(shí)間：2018-03-10 03:16

  本文選題：2型糖尿病　切入點(diǎn)：氧化應(yīng)激　出處：《山西醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過探討脂聯(lián)素對(duì)2型糖尿病(T2DM)大鼠骨髓中抗酒石酸酸性磷酸酶(TRAP)、骨保護(hù)素(OPG)與NF-κB受體活化配體(RANKL)表達(dá)水平的影響,結(jié)合脂聯(lián)素對(duì)骨髓微環(huán)境中糖基化終末產(chǎn)物(AGEs)及晚期氧化蛋白產(chǎn)物(AOPP)的影響,探討脂聯(lián)素是否可以通過調(diào)節(jié)氧化應(yīng)激水平對(duì)糖尿病骨代謝起一定的作用。方法:60只清潔級(jí)6周齡雄性SD大鼠隨機(jī)分為正常對(duì)照組(N組,n=18)及2型糖尿病模型組42只。正常對(duì)照組予普通飼料喂養(yǎng),2型糖尿病模型組先以高脂高糖飼料喂養(yǎng)8周,隨后按體重給予以一次性腹腔注射濃度為1%的鏈脲佐菌素(STZ)30mg/kg,以建立2型糖尿病大鼠模型。于建模成功后,選取造模成功的SD大鼠36只,隨機(jī)分為糖尿病組(DM組,n=18)及糖尿病脂聯(lián)素干預(yù)組(APN,n=18),APN組予皮下注射脂聯(lián)素10μg/kg·d干預(yù),普通組及糖尿病組大鼠給予等量生理鹽水。分別于干預(yù)后第4、8、12周末隨機(jī)選取6只符合標(biāo)準(zhǔn)的大鼠,麻醉后迅速取出雙側(cè)股骨及脛骨,提取一側(cè)股骨及脛骨骨髓液,經(jīng)離心分別留取骨髓細(xì)胞及骨髓上清液,另一側(cè)股骨測(cè)定骨密度后脫鈣包埋切片。ELISA、RT-PCR等方法檢測(cè)TRAP、RANKL、OPG、AOPP、AGEs,骨組織HE染色觀察骨小梁密度。結(jié)果:1.隨著飼養(yǎng)時(shí)間的延長(zhǎng),可見糖尿病組大鼠血糖水平明顯升高,而脂聯(lián)素干預(yù)組大鼠血糖水平較糖尿病組大鼠低(P0.05)。2.隨飼養(yǎng)時(shí)間延長(zhǎng),正常對(duì)照組大鼠骨組織免疫組化表達(dá)AGEs水平無明顯變化,DM組及APN組大鼠骨組織AGEs逐漸升高;三組間比較表明,4周時(shí),DM組大鼠較N組大鼠AGEs水平明顯升高(P0.05),脂聯(lián)素干預(yù)后,APN組大鼠較DM組明顯減低,8周及12周時(shí),DM組AGEs表達(dá)升高更顯著(P0.05),APN干預(yù)后較DM組降低(P0.05),且12周較8周兩組間AGEs下降幅度顯著增大。3.隨著飼養(yǎng)時(shí)間延長(zhǎng),N組大鼠骨髓上清液中AOPP無明顯變化,TRAP水平逐漸升高,DM組及APN組大鼠AOPP及TRAP水平均逐漸升高;4周時(shí),三組大鼠TRAP表達(dá)水平無明顯差異,DM組較N組大鼠AOPP水平明顯升高(P0.05),APN組較DM組無明顯差異。8周時(shí),與N組比較,DM組T2DM大鼠AOPP及TRAP水平明顯升高(P0.05),APN組大鼠骨髓上清液AOPP及TRAP水平較DM組明顯降低(P0.05),12周時(shí),兩組間AOPP及TRAP下降幅度增大(P0.05)。4.隨著喂養(yǎng)時(shí)間延長(zhǎng),各組大鼠骨髓細(xì)胞中OPG mRNA表達(dá)水平呈逐漸降低趨勢(shì),RANKL mRNA表達(dá)水平呈逐漸增高趨勢(shì)。4周時(shí),三組大鼠OPG mRNA、RANKL mRNA表達(dá)水平無明顯差異,8周時(shí),與N組比較,DM組大鼠骨髓細(xì)胞OPG mRNA表達(dá)水平顯著降低,RANKL mRNA表達(dá)水平顯著升高(P0.05),脂聯(lián)素干預(yù)后,APN組大鼠骨髓細(xì)胞OPG mRNA表達(dá)水平較DM組明顯升高,RANKL mRNA表達(dá)水平較DM組顯著降低,12周時(shí),兩組間比較變化幅度更加明顯。OPG/RANKL值與OPG mRNA表達(dá)水平變化趨勢(shì)一致(P0.001)。5.隨著時(shí)間延長(zhǎng),各組大鼠股骨頸BMD指標(biāo)均逐漸減低,以DM組明顯;4周時(shí),三組大鼠BMD無明顯差異,骨小梁結(jié)構(gòu)尚無明顯變化,12周時(shí),DM組較N組BMD水平明顯減低,骨小梁破壞嚴(yán)重,APN組較DM組BMD水平明顯升高,骨小梁結(jié)構(gòu)有所改善(P0.001)。結(jié)論:1.2型糖尿病大鼠糖代謝紊亂致骨髓微環(huán)境中糖基化終末產(chǎn)物生成增多,氧化應(yīng)激程度加重,可能使2型糖尿病大鼠破骨細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)通路中OPG/RANKL比值降低,結(jié)合TRAP顯著升高,表明破骨細(xì)胞分化生成增多,可促進(jìn)骨吸收過程,破壞骨小梁,影響骨質(zhì)量及骨密度,加速骨丟失。2.外源性脂聯(lián)素可能通過減少AGEs的生成,使糖尿病大鼠骨髓微環(huán)境中氧化應(yīng)激水平降低,升高OPG表達(dá),抑制RANKL表達(dá),降低2型糖尿病大鼠骨髓中破骨細(xì)胞分化,從而對(duì)骨代謝起正向的保護(hù)作用。
[Abstract]:Objective: To investigate adiponectin in type 2 diabetes mellitus (T2DM) rat tartrate resistant acid phosphatase in bone marrow (TRAP), osteoprotegerin (OPG) and NF- kappa B ligand receptor activation (RANKL) on the expression of adiponectin on bone marrow microenvironment, combined with advanced glycation end products (AGEs) and advanced oxidation protein products (AOPP) effect, investigate whether adiponectin can be regulated by oxidative stress plays a role in diabetic bone metabolism. Methods: 60 male 6 week old male SD rats were randomly divided into normal control group (group N, n=18) and type 2 diabetes model group with 42 rats in normal control group. Given common forage, type 2 diabetic model group with high-fat and high sugar diet for 8 weeks, then according to the weight given by intraperitoneal injection of 1% of the concentration of streptozotocin (30mg/kg, STZ) in type 2 diabetic rat model was established. After the success of Yu Jianmo, select the successful model of SD 36 rats were randomly divided into diabetic group (DM group, n=18) and diabetic group (APN, n=18, adiponectin), APN group received subcutaneous injection of adiponectin 10 g/kg - D group, normal group and diabetic group rats were given saline respectively. In the intervention after 4,8,12 weeks 6 rabbits were randomly selected for standard the rat, anesthesia was quickly removed after bilateral femur and tibia, femur and tibia bone marrow extraction side, bone marrow cells were collected by centrifugation and the supernatant of bone marrow, the other side of the femoral bone density after decalcified paraffin embedded.ELISA, RT-PCR, RANKL, OPG methods were used to detect TRAP, AOPP, AGEs, observe the trabecular bone density bone tissue HE staining. Results: 1. with the feeding time, blood glucose levels seen in diabetic group rats increased significantly, while the blood glucose level of adiponectin rats compared with diabetic group rats (P0.05.2.) with low feeding time, the normal control group rats from bone tissue 鐤粍鍖栬〃杈続GEs姘村鉤鏃犳槑鏄懼彉鍖，

本文編號(hào)：1591517