發(fā)布時(shí)間：2018-05-08 08:28

  本文選題：糖尿病腎病 + MIOX��； 參考：《中南大學(xué)》2014年碩士論文

【摘要】：研究背景 糖尿病腎病(diabetic nephropathy, DN)是糖尿病微血管并發(fā)癥的重要類型,也是引起終末期腎臟疾病(end stage renal disease, ESRD)的主要病因(約占40%-50%)。糖尿病腎病發(fā)病機(jī)制復(fù)雜,已有大量研究證實(shí)多元醇通路活性增高、糖基化終末產(chǎn)物形成增加、蛋白激酶C(protein kinase C,PKC)和RAS系統(tǒng)激活、GTP酶蛋白的活性變化、TGF-β-Smad信號通路的激活等細(xì)胞分子信號轉(zhuǎn)導(dǎo)途徑異常均參與了糖尿病腎病的發(fā)生與進(jìn)展,但其具體的發(fā)病機(jī)制尚不完全清楚。目前臨床常用針對上述單個(gè)因素及信號轉(zhuǎn)導(dǎo)分子的治療干預(yù)措施如PKC抑制劑等療效有限。因此深入開展糖尿病腎病分子發(fā)病機(jī)制的研究,尋找糖尿病腎病新的治療靶點(diǎn),一直是國內(nèi)外腎臟病學(xué)界研究領(lǐng)域的熱點(diǎn),具有重要的科學(xué)和社會(huì)意義。 傳統(tǒng)觀念認(rèn)為腎小球細(xì)胞損傷是糖尿病腎病的主要特征,但新近研究證實(shí)腎小管間質(zhì)損傷在糖尿病腎病中具有重要的意義,且腎小管間質(zhì)病變并不繼發(fā)于腎小球病變,而是糖尿病腎病的早期和原始特征。腎小管上皮細(xì)胞損傷可導(dǎo)致腎小管重吸收和分泌功能異常,促進(jìn)腎小管上皮細(xì)胞萎縮及腎間質(zhì)纖維化。最近研究證實(shí)肌醇耗竭在糖尿病腎病腎小管間質(zhì)損傷的過程中起著重要的作用,而腎臟特異性表達(dá)的肌醇加氧酶(Myo-inositol oxygenase, MIOX)主要表達(dá)在腎組織腎小管上皮細(xì)胞內(nèi),是組織中肌醇代謝的關(guān)鍵調(diào)節(jié)酶。體外實(shí)驗(yàn)表明高葡萄刺激可上調(diào)腎小管上皮細(xì)胞內(nèi)肌醇加氧酶的表達(dá),并產(chǎn)生促進(jìn)活性氧(reactive oxygen species, ROS)生成及纖維連接蛋白(fibronectin, FN)表達(dá)的效應(yīng)�；谝陨戏治�,我們認(rèn)為MIOX在糖尿病腎病患者腎組織內(nèi)表達(dá)上調(diào),并可能與糖尿病腎病腎小管上皮細(xì)胞氧化損傷及間質(zhì)纖維化密切相關(guān)。故本試驗(yàn)中我們通過檢測MIOX在糖尿病腎病患者腎活檢組織及血液、尿液中的表達(dá),并分析其余臨床指標(biāo)之間的相關(guān)性,旨在為進(jìn)一步研究糖尿病腎病的發(fā)病機(jī)制及治療提供新的思路與靶點(diǎn)。 第一章糖尿病腎病患者腎組織中MIOX的表達(dá)及其與臨床指標(biāo)相關(guān)性分析 目的：觀察MIOx在糖尿病腎病患者腎組織中的表達(dá),并分析其與臨床各指標(biāo)之間的關(guān)系。 方法：隨機(jī)選取經(jīng)臨床和腎臟病理活檢確診為糖尿病腎病的患者15人及輕微病變的患者(原發(fā)性腎病)15人。采用HE, PASM, Masson,PAS染色觀察腎臟病理改變,免疫組化、免疫熒光檢測MIOX表達(dá)病變,并使用SPSS17.0分析糖尿病腎病患者腎組織內(nèi)MIOX相對表達(dá)程度與臨床指標(biāo)間的相關(guān)性。 結(jié)果：腎臟病理顯示糖尿病腎病患者腎活檢組織腎小球體積增大,部分出現(xiàn)結(jié)節(jié)性腎小球硬化,出現(xiàn)典型的Kimmelstiel-Wilson結(jié)節(jié),腎小球系膜基質(zhì)的增生,基底膜的增厚、僵硬；腎小管片狀萎縮并部分代償性擴(kuò)張,腎間質(zhì)的片狀纖維化改變。而對照組為輕微病變患者,腎小球部分基本正常,系膜細(xì)胞及系膜基質(zhì)都無明顯增生,腎小管間質(zhì)無明顯纖維化,血管無明顯異常。糖尿病腎病患者的腎小管間質(zhì)損傷評分明顯比對照組高。免疫組化實(shí)驗(yàn)結(jié)果提示：MIOX主要表達(dá)在腎小管上皮細(xì)胞胞漿內(nèi)。免疫熒光試驗(yàn)結(jié)果提示：MIOX主要表達(dá)在腎小管上皮細(xì)胞的胞漿及腎間質(zhì)內(nèi)。與非糖尿病腎病輕微病變相比,糖尿病腎病患者腎小管細(xì)胞內(nèi)MIOX表達(dá)顯著上調(diào)(P0.05)。相關(guān)性分析表明MIOX表達(dá)強(qiáng)度與糖尿病腎病患者腎小管間質(zhì)損傷評分、血清肌酐、血糖、糖化血紅蛋白水平呈正相關(guān)。 結(jié)論：糖尿病腎病患者腎組織MIOX表達(dá)水平明顯上升,MIOX可能參與糖尿病腎病腎小管間質(zhì)損傷。 第二章糖尿病腎病患者血液及尿液中MIOX的表達(dá)及其與臨床指標(biāo)的相關(guān)性分析 目的：觀察糖尿病腎病患者血清及尿液中MIOX的表達(dá),分析其與臨床指標(biāo)的關(guān)系。 方法：隨機(jī)選取臨床診斷為糖尿病腎病患者30人,非糖尿病腎病慢性腎臟疾病患者30人,單純糖尿病患者30人,正常對照組30人。采用ELISA法檢測外周血及尿液中間的MIOX表達(dá)。 結(jié)果：MIOX檢測結(jié)果顯示：糖尿病腎病組,非糖尿病腎病組,糖尿病組,及正常對照組血清MIOX的水平分別為:2050.4士160.7、1069.2±96.23、619.3±64.1、49.47±3.3pg/ml。糖尿病腎病組,非糖尿病腎病組,糖尿病組,及正常對照組尿液MIOX的水平分別為：2675.3±241.1、1427.2±146.3、775.0±80.3、73.5±4.8pg/ml。糖尿病腎病患者血,尿MIOX水平明顯高于非糖尿病腎病組、單純糖尿病患者組及正常對照組,差異具有統(tǒng)計(jì)學(xué)差異(p0.01),相關(guān)性分析發(fā)現(xiàn)糖尿病腎病患者血清及尿液MIOX表達(dá)水平與血清肌酐、血糖、糖化血紅蛋白水平呈正相關(guān)。 結(jié)論：糖尿病腎病患者外周血及尿液中MIOX的表達(dá)水平明顯上調(diào)。
[Abstract]:Research background
Diabetic nephropathy (DN) is an important type of diabetic microvascular complication and is also the main cause of end stage renal disease (ESRD). The pathogenesis of diabetic nephropathy is about 40%-50%. The pathogenesis of diabetic nephropathy is complex. A large number of studies have confirmed that the activity of polyol pathway is increased and the end products of glycosylation are increased. In addition, the activation of protein kinase C (protein kinase C, PKC) and RAS system, the changes in the activity of GTP enzyme protein and the activation of the TGF- beta -Smad signaling pathway are all involved in the development and progression of diabetic nephropathy, but the specific pathogenesis is not completely clear. The therapeutic intervention of number transducers, such as PKC inhibitors, is limited. Therefore, it is of great scientific and social significance to develop the molecular pathogenesis of diabetic nephropathy and find new therapeutic targets for diabetic nephropathy, which has always been a hot spot in the field of Nephrology at home and abroad.
Traditionally, glomerular cell damage is the main feature of diabetic nephropathy. However, recent studies have confirmed that renal tubulointerstitial damage is of great significance in diabetic nephropathy. Renal tubulointerstitial lesions are not secondary to glomerular lesions, but are early and original characteristics of diabetic nephropathy. Renal tubular epithelial cell damage can lead to kidney disease. The renal tubular epithelial cell atrophy and renal interstitial fibrosis are promoted by the abnormal reabsorption and secretory function of the tubules. Recent studies have shown that inositol depletion plays an important role in the process of renal tubulointerstitial damage in diabetic nephropathy, and the renal specific expression of inositol oxygenase (Myo-inositol oxygenase, MIOX) is mainly expressed in renal tissue. In vitro, it is the key regulator of the metabolism of inositol in the tissue. In vitro experiments show that high grape stimulation can increase the expression of inositol oxygenase in renal tubular epithelial cells and produce the effect of reactive oxygen species (ROS) generation and fibronectin (fibronectin, FN) expression. Based on the above analysis, we believe that M The expression of IOX is up-regulated in the renal tissue of patients with diabetic nephropathy and may be closely related to the oxidative damage and interstitial fibrosis of renal tubular epithelial cells in diabetic nephropathy. In this study, we detected the expression of MIOX in renal biopsy tissue, blood and urine in patients with diabetic nephropathy, and analyzed the correlation between the other clinical indicators. It will provide new ideas and targets for further study on the pathogenesis and treatment of diabetic nephropathy.
Chapter 1 expression of MIOX in renal tissue of patients with diabetic nephropathy and its correlation with clinical indicators
Objective: To observe the expression of MIOx in renal tissue of patients with diabetic nephropathy and analyze its relationship with clinical indicators.
Methods: 15 patients with diabetic nephropathy and 15 patients with mild pathological changes (primary nephrosis) were randomly selected by clinical and renal biopsy. HE, PASM, Masson, PAS staining were used to observe renal pathological changes, immunohistochemistry, immunofluorescence detection of MIOX expression, and SPSS17.0 analysis of MI in renal tissue of diabetic nephropathy patients. The correlation between the relative expression level of OX and clinical indicators.
Results: renal pathology showed that the glomerular volume increased in renal biopsy tissue, nodular glomerulosclerosis, typical Kimmelstiel-Wilson nodules, glomerular mesangial matrix hyperplasia, basilar membrane thickening, stiffness, renal tubuloatrophy and partial compensatory dilation, and renal interstitial fibrosis. In the control group, the glomerular part was basically normal, the mesangial cells and mesangial matrix had no obvious hyperplasia, there was no obvious fibrosis in the tubulointerstitium and no obvious abnormal blood vessels. The renal tubulointerstitial damage score of diabetic nephropathy patients was significantly higher than that of the control group. The immunohistochemical results suggested that MIOX was mainly expressed in the kidneys. The results of immunofluorescence test showed that MIOX was mainly expressed in the cytoplasm and renal interstitium of renal tubular epithelial cells. Compared with non diabetic nephropathy, the expression of MIOX in renal tubule cells in diabetic nephropathy patients was significantly up-regulated (P0.05). The correlation analysis showed that the expression intensity of MIOX was smaller than that of diabetic nephropathy. There was a positive correlation between tubulointerstitial injury score, serum creatinine, blood sugar and glycosylated hemoglobin level.
Conclusion: the expression level of MIOX in renal tissue of patients with diabetic nephropathy increases significantly, and MIOX may be involved in tubulointerstitial injury in diabetic nephropathy.
The second chapter is about the expression of MIOX in blood and urine of patients with diabetic nephropathy and its correlation with clinical indicators.
Objective: To observe the expression of MIOX in serum and urine of patients with diabetic nephropathy and analyze its relationship with clinical indicators.
Methods: 30 patients with diabetic nephropathy, 30 patients with chronic renal disease in non diabetic nephropathy, 30 patients with simple diabetes and 30 normal controls were randomly selected. The MIOX expression in the middle of peripheral blood and urine was detected by ELISA.
Results: the results of MIOX detection showed that the levels of serum MIOX in diabetic nephropathy group, non diabetic nephropathy group, diabetes group and normal control group were 2050.4 160.71069.2 + 96.23619.3 + 64.1,49.47 + 3.3pg/ml. diabetic nephropathy group, non diabetic nephropathy group, diabetes group, and normal control group urine MIOX level were 2675, respectively: The serum and urine MIOX levels of.3 + 241.11427.2 + 146.3775.0 + 80.3,73.5 + 4.8pg/ml. diabetic nephropathy patients were significantly higher than those of non diabetic nephropathy group. The difference was statistically significant (P0.01). The correlation analysis found the serum and urine MIOX expression level and serum creatinine, blood glucose, and blood glucose in patients with diabetic nephropathy. The level of glycosylated hemoglobin was positively correlated.
Conclusion: the expression level of MIOX in peripheral blood and urine of diabetic nephropathy patients is significantly increased.

【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R587.2;R692.9

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 崔佳;竇京濤;;糖尿病伴高血糖腎傷害的特征、識別與臨床處理[J];實(shí)用糖尿病雜志;2013年04期

2 張欣;;miRNA與糖尿病腎病纖維化的關(guān)系[J];牡丹江醫(yī)學(xué)院學(xué)報(bào);2014年04期

3 嚴(yán)小倩;魯盈;;雷公藤多苷治療糖尿病腎病的利與弊[J];中國中西醫(yī)結(jié)合腎病雜志;2014年07期

4 吳歌;賈曉媛;黃博;高丹;;代謝綜合征與中青年慢性腎臟病的相關(guān)性研究[J];中國全科醫(yī)學(xué);2014年02期

5 張艷;任榮;梁新華;張蕾;;早期糖尿病腎病泛素-核糖體融合蛋白52和尿微量清蛋白關(guān)系研究[J];中國全科醫(yī)學(xué);2014年26期

6 趙龍;關(guān)廣聚;;糖尿病腎病發(fā)病機(jī)制研究進(jìn)展[J];中華腎臟病雜志;2013年07期

7 邵科晶;朱寶;袁航;許亞豐;朱慧;王菲;;~(99m)Tc-DTPA腎小球?yàn)V過率測定在診斷糖尿病早期腎損害中的價(jià)值[J];中華臨床醫(yī)師雜志(電子版);2013年14期

8 Maria L Gonzalez Suarez;David B Thomas;Laura Barisoni;Alessia Fornoni;;Diabetic nephropathy:Is it time yet for routine kidney biopsy?[J];World Journal of Diabetes;2013年06期

9 沈潔;婁霞英;邢玉波;馬江波;宋迎香;張宇律;張煒;華燕吟;;糖尿病患者對糖尿病腎病認(rèn)知水平及影響因素研究[J];浙江預(yù)防醫(yī)學(xué);2014年06期

10 張艷;任榮;張蕾;;早期糖尿病腎病患者血糖、腎小球率過濾、NAG酶與尿UBA52的關(guān)系研究[J];新疆醫(yī)科大學(xué)學(xué)報(bào);2014年07期

相關(guān)博士學(xué)位論文 前5條

1 辛欣;五甲基槲皮素抗糖尿病大鼠腎臟纖維化的作用及機(jī)制研究[D];華中科技大學(xué);2013年

2 邵明龍;LDR與FGF21聯(lián)用對2型糖尿病腎臟保護(hù)作用研究[D];吉林大學(xué);2014年

3 孫立;黃芪甲苷對高糖所致人腎小球系膜細(xì)胞損傷的保護(hù)作用及其調(diào)控NADPH氧化酶/ROS/Akt/NF-κB信號通路的機(jī)制研究[D];安徽醫(yī)科大學(xué);2014年

4 趙艷艷;鄭州地區(qū)城市居民2型糖尿病危險(xiǎn)因素分析及SIRT1和FOXO1單核苷酸多態(tài)性與糖尿病腎病的關(guān)聯(lián)研究[D];鄭州大學(xué);2014年

5 陽石坤;普羅布考通過P66Shc保護(hù)糖尿病腎病腎小管間質(zhì)損傷的作用及機(jī)制研究[D];中南大學(xué);2014年

相關(guān)碩士學(xué)位論文 前10條

1 楊瑞鳳;活性維生素D_3對糖尿病大鼠腎組織BMP-7、TGF-β1表達(dá)的影響[D];鄭州大學(xué);2013年

2 王鳳玲;小檗堿對高脂加鏈脲佐菌素誘導(dǎo)的糖尿病腎病大鼠腎臟的保護(hù)作用及其對G蛋白偶聯(lián)受體激酶的調(diào)控[D];安徽醫(yī)科大學(xué);2013年

3 沃冠群;木犀草素通過激活Nrf2途徑在糖尿病腎病進(jìn)展中的治療作用[D];南京中醫(yī)藥大學(xué);2013年

4 宋盼愛;PKCδ對高葡萄糖誘導(dǎo)的HK-2細(xì)胞p66Shc磷酸化及線粒體轉(zhuǎn)位的影響[D];中南大學(xué);2013年

5 楊玉艷;Rap1b通過GSK-3β/β-catenin信號抑制高糖誘導(dǎo)的小管上皮細(xì)胞凋亡[D];中南大學(xué);2013年

6 季家舉;嗜酸氧化亞鐵硫桿菌異二硫化物還原酶HdrC亞基的表達(dá)純化及定點(diǎn)突變[D];中南大學(xué);2012年

7 張晶晶;2型糖尿病大血管并發(fā)癥中血漿微小RNA表達(dá)譜的研究[D];華中科技大學(xué);2013年

8 鄭書香;肌醇氧化酶在大腸桿菌中的高效表達(dá)及其催化生產(chǎn)葡萄糖醛酸的特性研究[D];浙江大學(xué);2013年

9 何升林;血清miRNA在糖尿病腎病中預(yù)警作用研究[D];蘭州大學(xué);2014年

10 章峻鈞;2型糖尿病患者微血管病變的影響因素研究[D];廣西醫(yī)科大學(xué);2014年

，

本文編號：1860684