本文選題：糖尿病 + 腎病　； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:糖尿病腎病(Diabetic Kidney Disease,DKD)是糖尿病(Diabetes Mellitus,DM)常見的微血管并發(fā)癥之一,晚期可以進(jìn)展至終末腎功能衰竭(end-stage renal failure,ESRF),導(dǎo)致患者死亡,成為嚴(yán)重危害人類生活質(zhì)量并威脅人類生命的疾病。近年來研究發(fā)現(xiàn)他汀類藥物除突出的降脂作用以外,其對糖尿病患者腎臟的非降脂的保護(hù)作用受到越來越多的關(guān)注。一氧化氮(nitric oxide,NO)作為人體內(nèi)重要的血管舒張因子,可以擴(kuò)張腎小球入球小動脈及出球小動脈,降低腎血管阻力,改善腎小球濾過率和腎臟血流灌注,以維持腎臟良好的血流動力學(xué)狀態(tài)及血管張力。本實驗通過觀察阿托伐他汀對2型糖尿病大鼠腎組織中內(nèi)皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)表達(dá)水平、NO含量等的影響,探討其是否會通過干預(yù)eNOS-NO通路減慢糖尿病腎病進(jìn)展,從而為他汀類藥物對臨床糖尿病腎臟保護(hù)提供理論依據(jù)。方法:選取160-200g健康清潔級雌性SD大鼠54只,隨機(jī)分為兩組,一組為正常對照組(NC組),另一組為高糖高脂喂養(yǎng)同時聯(lián)合小劑量鏈脲佐菌素(STZ)(30 mg/kg)一次性腹腔注射建立的2型糖尿病大鼠模型組。于實驗第10w末稱取大鼠體重(BW),取下腔靜脈血標(biāo)本檢測空腹胰島素(FINS)、空腹血糖(FBG)、總膽固醇(TC)和甘油三酯(TG),并根據(jù)FBG和FINS計算胰島素抵抗指數(shù)(HOMA-IR)。之后將2型糖尿病大鼠隨機(jī)分為糖尿病組(DM組)和阿托伐他汀干預(yù)組(A組)。每周監(jiān)測血糖、BW。實驗第21w末稱取各組大鼠體重(BW),取大鼠下腔靜脈血測定FINS、FBG、TC、TG、糖化血紅蛋白(Hb A1c)、谷丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST)、肌酐(SCr)、尿素(UREA),計算胰島素抵抗指數(shù)(HOMA-IR);取出大鼠腎臟后,去除被膜、脂肪等腎臟以外組織后稱取雙腎重量(KW),計算腎臟指數(shù)(Kidney Index,KI=KW/BW);腎臟組織石蠟切片行HE染色、PAS染色觀察大鼠腎臟病理改變;取部分腎皮質(zhì)勻漿以硝酸還原法測定NO含量;應(yīng)用real-time PCR檢測腎皮質(zhì)eNOS m RNA含量;免疫組織化學(xué)染色檢測eNOS在腎皮質(zhì)中的表達(dá)。應(yīng)用SPSS 21.0統(tǒng)計軟件進(jìn)行數(shù)據(jù)分析,數(shù)據(jù)描述用均數(shù)±標(biāo)準(zhǔn)差(`x±s)表示。兩組間樣本均數(shù)比較采用獨立樣本t檢驗,多組間樣本均數(shù)的多重比較采用LSD、SNK-q、Tamhane檢驗。P0.05認(rèn)為差異具有統(tǒng)計學(xué)意義。結(jié)果:1 BW變化實驗第10w末,糖尿病大鼠的BW明顯高于NC組大鼠(P0.05)。第21w末,與NC組比較,DM組大鼠和A組大鼠的BW均明顯下降(P0.01);DM組大鼠和A組大鼠相比,體重?zé)o顯著差異(P0.05)。2血生化指標(biāo)檢測實驗第10w末,糖尿病大鼠的FBG、TG、TC、FINS均明顯高于同期NC組大鼠(P0.01)。第21w末,DM組和A組大鼠FINS、FBG、TC、TG、Hb A1c與同期NC組大鼠相比均明顯升高(P0.01,P0.05),而DM組與A組大鼠上述指標(biāo)差異無統(tǒng)計學(xué)意義(P0.05);DM組和A組大鼠UREA水平均高于同期NC組大鼠(P0.05),A組大鼠和DM組大鼠相比,UREA水平差異無統(tǒng)計學(xué)意義(P0.05);3組大鼠SCr、ALT、AST均無顯著差異(P0.05)。3胰島素敏感性測定實驗第10w末,糖尿病大鼠的HOMA-IR顯著高于同期NC組(P0.01),證實糖尿病大鼠存在胰島素抵抗,為2型糖尿病大鼠。第21周末,DM組、A組大鼠HOMA-IR仍顯著高于同期NC組大鼠(P0.01),DM組與A組相比較HOMA-IR無顯著差異(P0.05)。4腎臟指數(shù)測定實驗第21w末,DM組和A組大鼠的雙腎重量(KW)和腎臟指數(shù)(Kidney Index,KI=KW(mg)/BW(g))均明顯高于NC組大鼠(P0.01),A組大鼠上述指標(biāo)較DM組大鼠均降低,差異有統(tǒng)計學(xué)意義(P0.01)。5腎臟病理改變NC組大鼠腎小球及腎小管形態(tài)正常。DM組大鼠腎小球肥大,系膜區(qū)擴(kuò)張,基底膜不規(guī)則增厚,PAS陽性物質(zhì)沉積增多。部分腎小管出現(xiàn)細(xì)胞空泡樣變性,管腔變窄。A組大鼠腎臟病變較DM組稍有減輕,表現(xiàn)為腎小球輕度增大,基底膜輕微增厚,PAS陽性物質(zhì)沉積有所減少,偶可見腎小管細(xì)胞空泡樣變性及管腔變窄。6腎皮質(zhì)NO含量檢測DM組、A組大鼠腎皮質(zhì)NO含量低于同期NC組大鼠(P0.01),DM組大鼠腎皮質(zhì)NO含量低于A組大鼠,差異有統(tǒng)計學(xué)意義(P0.01)。7腎皮質(zhì)eNOS免疫組化染色胞漿中呈棕黃色染色顆粒者為eNOS陽性表達(dá)。DM組和A組大鼠腎皮質(zhì)eNOS表達(dá)均顯著低于NC組大鼠(P0.01,P0.05),A組大鼠腎皮質(zhì)eNOS表達(dá)高于DM組大鼠,差異有統(tǒng)計學(xué)意義(P0.05)。8腎皮質(zhì)eNOS m RNA real-time PCR結(jié)果與NC組相比,DM組和A組大鼠腎皮質(zhì)中eNOS m RNA含量均降低(P0.01);其中A組大鼠eNOS m RNA含量高于DM組大鼠(P0.01)。結(jié)論:1在高糖高脂飲食的基礎(chǔ)上聯(lián)合小劑量鏈脲佐菌素一次性腹腔注射能夠成功建立2型糖尿病大鼠模型,隨著病程的進(jìn)展,其腎臟組織學(xué)改變基本具備人類2型糖尿病腎病的典型病理特征。2與正常對照組相比,糖尿病大鼠腎皮質(zhì)eNOS表達(dá)下降,腎皮質(zhì)中NO含量降低,與DM組相比,應(yīng)用阿托伐他汀對FBG、Hb A1C、HOMA-IR等雖無顯著影響,但其可以明顯改善腎臟病理學(xué)變化,上調(diào)腎皮質(zhì)中eNOS的表達(dá),增加NO合成,提示阿托伐他汀對糖尿病大鼠腎臟可能具有非降脂的保護(hù)作用。
[Abstract]:Objective: Diabetic Kidney Disease (DKD) is one of the common microvascular complications of diabetes (Diabetes Mellitus, DM). In the late stage, it can advance to terminal renal failure (end-stage renal failure, ESRF), causing death and becoming a serious threat to human life and threat to human life. In recent years, research has been found. In addition to the protruding effect of lipid lowering, statins have attracted more and more attention to the non lipid lowering effect of kidney in diabetic patients. Nitric oxide (NO), as an important vascular diastolic factor in the human body, can dilate the glomerular arteriole and arteriole, reduce renal vascular resistance and improve glomerular filtration. The effect of atorvastatin on the expression of endothelial nitric oxide synthase (eNOS) and the content of NO in renal tissue of type 2 diabetic rats was investigated to investigate whether it could interfere with eNOS-NO through the observation of the effect of atorvastatin on the expression of endothelial nitric oxide synthase (oxide synthase, eNOS) in the renal tissue of type 2 diabetic rats. The progress of diabetic nephropathy was slowed down to provide a theoretical basis for statins to protect the kidney from clinical diabetes. Methods: 54 healthy and clean female SD rats of 160-200g were randomly divided into two groups, one group was the normal control group (group NC) and the other group was combined with low dose streptozotocin (STZ) (30 mg/kg) at the same time. The rat model group of type 2 diabetic rats was established by intraperitoneal injection. The rat body weight (BW) was measured at the end of the experiment at the end of the experiment. The fasting insulin (FINS), fasting blood glucose (FBG), total cholesterol (TC) and triglyceride (TG) were measured in the inferior vena cava blood samples, and the insulin resistance index (HOMA-IR) was calculated according to FBG and FINS. Then, type 2 diabetic rats were randomly divided into sugars. Urine disease group (group DM) and atorvastatin intervention group (group A) monitoring blood sugar weekly, BW. experiment 21W end 21W was called group rats weight (BW), the rat inferior vena cava blood was taken to determine FINS, FBG, TC, TG, glycosylated hemoglobin (Hb A1c), glutamic aminotransferase (ALT), cereal transaminase, creatinine, urea and insulin resistance index. After removing the kidneys of the rats, the kidney index (KW) was removed and the kidney index (Kidney Index, KI=KW/BW) was calculated after removal of the membrane, fat and other tissues outside the kidney. The renal tissue paraffin sections were stained with HE, and the renal pathological changes were observed by PAS staining. Some renal cortex homogenates were measured with nitric acid reduction method and NO content was measured. Real-time PCR was used to detect eNOS m R. ENOS of real-time PCR was used to detect eNOS m R. NA content; immunohistochemical staining was used to detect the expression of eNOS in the renal cortex. The data was analyzed with SPSS 21 statistical software, and the data description was expressed with mean mean standard deviation (`x + s). The average number of samples between the two groups was compared with the independent sample t test. The multiple comparison of the average numbers of the multiple groups adopted LSD, SNK-q, and Tamhane test.P0.05. The results were statistically significant. Results: the BW of diabetic rats was significantly higher than that in group NC (P0.05) at the end of the 1 BW change experiment. At the end of 21W, the BW of both DM and A rats decreased significantly (P0.01) compared with those in the NC group, and there was no significant difference in weight between the rats of the DM group and the rats. TG, TC and FINS were significantly higher than that of NC rats in the same period (P0.01). At the end of 21W, FINS, FBG, TC, TG, and A groups were significantly higher than those in the same period. There was no significant difference in UREA level in the group of rats (P0.05), and there was no significant difference between the 3 groups of rats SCr, ALT and AST (P0.05).3 insulin sensitivity test 10W, and the HOMA-IR in diabetic rats was significantly higher than that of the NC group (P0.01) in the same period. It was confirmed that the diabetic rats had insulin resistance, which was type 2 diabetic rats. Twenty-first weekend, DM group, and rats were the rats. A-IR was still significantly higher than that of NC group (P0.01), and there was no significant difference in HOMA-IR between group DM and A group (P0.05).4 kidney index test, and both kidney weight (KW) and kidney index in DM and A group were significantly higher than that of group rats. There were statistically significant (P0.01).5 renal pathological changes in group NC, glomeruli and renal tubules in normal.DM rats, glomerular hypertrophy, mesangial region dilation, irregular thickening of basement membrane, and increased PAS positive substance deposition. Some renal tubules appeared vacuolated degeneration of cells and renal lesions in group.A group were slightly lessened than those in the DM group, showing kidneys. Slight enlargement of small ball, slight thickening of basement membrane and decrease of PAS positive substance deposition, NO content in renal cortex of renal tubule cells and narrowing of.6 in group DM, NO content in renal cortex of A group was lower than that of NC group (P0.01) in A group, and the content of NO content in renal cortex of DM group was lower than that of A group, the difference was statistically significant (P0.01) kidney The expression of eNOS expression in.DM group and A group was significantly lower than that of NC group (P0.01, P0.05) in.DM group and A group. The expression of eNOS expression in renal cortex of A group was higher than that of DM group, and the difference was statistically significant (P0.05) compared with that of the group of NC. The content of eNOS m RNA in renal cortex of group and A rats decreased (P0.01), and eNOS m RNA content in group A rats was higher than that of group DM rats (P0.01). Conclusion: 1 a one-time injection of small dose streptozotocin combined with small dose of streptozotocin on the basis of high glucose and high fat diet can successfully establish a model of type 2 diabetic rats. With the progress of the course of disease, the kidney histology changes The typical pathological features of human type 2 diabetic nephropathy had a typical pathological feature.2, compared with the normal control group, the expression of eNOS in the renal cortex of diabetic rats decreased and the NO content in the renal cortex decreased. Compared with the DM group, the use of atorvastatin had no significant influence on FBG, Hb A1C, HOMA-IR and so on, but it could obviously improve the renal pathological changes and up the renal cortex. The expression of eNOS increased NO synthesis, suggesting that atorvastatin may protect the kidneys of diabetic rats from lipid lowering.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.2;R692.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Prevention of diabetic microangiopathy by prophylactic transplant of mobilized peripheral blood mononuclear cells[J];Acta Pharmacologica Sinica;2007年01期

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本文編號：1871268