本文選題：鋅-α2-糖蛋白 + 造影劑腎病��； 參考：《大連醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：造影劑腎�。╟ontrast-induced nephropathy，CIN）已成為院內(nèi)獲得性急性腎損傷的常見(jiàn)病因，而臨床上尚無(wú)敏感的檢測(cè)指標(biāo)及有效的治療手段。如何早期診斷對(duì)預(yù)防CIN極其重要。而血肌酐在CIN診斷的敏感性及特異性方面存在缺陷，故發(fā)現(xiàn)一種能在CIN早期即可檢測(cè)到的、敏感的、特異的生物學(xué)標(biāo)記物對(duì)預(yù)防CIN的發(fā)生具有重要意義。鋅-α2-糖蛋白（Zinc-α2-glycoprotein，ZAG）是正常腎小管上皮細(xì)胞表達(dá)的糖蛋白。本研究通過(guò)檢測(cè)經(jīng)皮冠狀動(dòng)脈介入治療（percutaneous coronary intervention，PCI）患者尿中ZAG水平的變化情況，以及含碘造影劑（iodine contrast media，CM）誘導(dǎo)大鼠腎小管上皮細(xì)胞（NRK-52E）ZAG表達(dá)與細(xì)胞凋亡關(guān)系，來(lái)評(píng)價(jià)尿中ZAG水平對(duì)CIN的早期預(yù)測(cè)作用，并初步探討ZAG在CIN發(fā)病機(jī)制中的作用。 方法：選取我院心內(nèi)科行PCI術(shù)患者（包括不穩(wěn)定型心絞痛、陳舊性及急性心肌梗死的患者）10例，所有患者獲知情同意。應(yīng)用的CM均為低滲非離子型單體造影劑碘佛醇370。造影劑注射方法均為Judkins法行冠脈造影，并根據(jù)術(shù)中所見(jiàn)冠狀動(dòng)脈狹窄程度行冠脈介入治療。入選標(biāo)準(zhǔn)：所有患者均行PCI術(shù)，術(shù)前采取靜脈血，檢測(cè)血肌酐，并根據(jù)MDRD公式計(jì)算eGFR（estimated glomerularfiltration rate估測(cè)腎小球?yàn)V過(guò)率）≥90ml/min·1.73m2。排除標(biāo)準(zhǔn)：1）慢性腎臟�。�2）各種原因所致的急性腎損傷，如近期腎毒性藥物的應(yīng)用、腎前性缺血等；3）合并糖尿病、惡性腫瘤、高脂血癥的患者；4）7天內(nèi)接受過(guò)靜脈造影的患者。留取患者術(shù)前及術(shù)后2h、24h尿液標(biāo)本，-40℃冰箱保存，檢測(cè)尿肌酐水平，用ELISA法檢測(cè)尿中ZAG濃度，尿ZAG水平用尿ZAG濃度與尿肌酐比值（mg/g）表示，同時(shí)采取造影前靜脈血標(biāo)本，檢測(cè)生化指標(biāo)包括：血肌酐、血尿素、膽固醇、低密度脂蛋白、甘油三酯。同時(shí)體外培養(yǎng)NRK-52E細(xì)胞，用不同碘濃度（25、50、100、140mgI/ml）的碘佛醇作用細(xì)胞2h，，用碘濃度為100mgI/ml的碘佛醇作用細(xì)胞2h、4h、6h。應(yīng)用流式Annexin V-FITC/PI雙染色法檢測(cè)不同碘濃度及不同時(shí)間誘導(dǎo)下細(xì)胞凋亡率，同時(shí)分別收集各組細(xì)胞總蛋白，應(yīng)用Western blot法檢測(cè)各組細(xì)胞ZAG蛋白表達(dá)情況。 結(jié)果：10例患者中，男6例，女4例，平均年齡58.40±6.50歲，術(shù)前尿素氮5.98±0.67mmol/L，膽固醇3.97±0.76mmol/L，甘油三酯1.39±0.34mmol/L，低密度脂蛋白2.09±0.57mmol/L，造影劑用量101.00±15.24ml，血肌酐63.83±9.65umol/L，計(jì)算eGFR為104.07±11.08ml/min·1.73m2。術(shù)前尿中ZAG水平為22.85±11.10mg/g。術(shù)后2h、24h尿中ZAG水平分別為67.44±44.94mg/g、32.24±18.55mg/g，與術(shù)前尿中ZAG水平相比，術(shù)后2h尿中ZAG水平較前升高，差異有統(tǒng)計(jì)學(xué)意義（p0.05）。術(shù)后24h雖較2h有下降，但仍高于術(shù)前，差異無(wú)統(tǒng)計(jì)學(xué)意義（p0.05）。各組尿ZAG水平與各項(xiàng)主要指標(biāo)均無(wú)相關(guān)性。體外培養(yǎng)的NRK-52E細(xì)胞，用不同碘濃度的碘佛醇作用細(xì)胞2h，及碘濃度為100mgI/ml的碘佛醇作用細(xì)胞不同時(shí)間后，應(yīng)用流式Annexin V-FITC/PI雙染色法檢測(cè)細(xì)胞凋亡率，與對(duì)照組相比，細(xì)胞凋亡率呈碘濃度(P0.05，n=3)和時(shí)間依賴(lài)性(P0.05，n=3)增加，應(yīng)用Western blot法檢測(cè)ZAG蛋白表達(dá)水平亦呈碘濃度(P0.05，n=3)和時(shí)間依賴(lài)性(P0.05，n=3)增加。線(xiàn)性回歸分析提示ZAG蛋白表達(dá)水平與細(xì)胞凋亡率呈正相關(guān)。 結(jié)論：1、尿中ZAG水平升高可能反應(yīng)含碘造影劑所致的早期腎損傷。 2、CM誘導(dǎo)的NRK-52E細(xì)胞中ZAG蛋白表達(dá)與細(xì)胞凋亡水平增加相關(guān)。
[Abstract]:Objective: contrast-induced nephropathy (CIN) has become a common cause of nosocomial acute renal injury, but there is no sensitive detection index and effective treatment. The early diagnosis is very important for the prevention of CIN. However, the sensitivity and specificity of serum creatinine in the diagnosis of CIN have been found to be defective. The sensitive, specific biological markers that can be detected at the early stage of CIN are important for the prevention of the occurrence of CIN. Zinc alpha 2- glycoprotein (Zinc- alpha 2-glycoprotein, ZAG) is a glycoprotein expressed in normal tubular epithelial cells. This study was conducted by detecting percutaneous coronary artery intervention (percutaneous coronary intervention, PCI). The changes in the level of ZAG in the urine and the iodine contrast media (CM) induced the relationship between the expression of NRK-52E ZAG and apoptosis in rat renal tubular epithelial cells (NRK-52E), to evaluate the early prediction of the level of ZAG in urine on CIN, and to explore the role of ZAG in CIN pathogenesis.
Methods: 10 patients (including unstable angina pectoris, obsolete and acute myocardial infarction) were selected in the Department of Cardiology of our hospital. All patients received informed consent. All of the patients were used in the CM of the low permeability nonionic single contrast agent iormol 370. contrast agent. The coronary angiography was performed by the Judkins method, and the coronary artery was observed according to the coronary artery in the operation. The degree of stenosis was treated with coronary intervention. Admission criteria: all patients underwent PCI surgery, preoperative venous blood was taken, serum creatinine was detected, and eGFR (estimated GlomerularFiltration Rate estimated glomerular filtration rate) > 90ml/min. 1.73m2. exclusion criteria were calculated according to the MDRD formula: 1) slow kidney disease; 2) acute renal injury caused by various causes, such as Recent use of nephrotoxic drugs, prerenal ischemia, etc.; 3) patients with diabetes, malignant tumor, hyperlipidemia, and hyperlipidemia; 4) patients who had received intravenous angiography within 7 days. 2h, 24h urine specimens, -40 centigrade refrigerators were preserved, urine creatinine levels were detected, ZAG concentration in urine was detected by ELISA, ZAG concentration in urine ZAG level and urinary ZAG concentration and urine ZAG level were measured. The urine creatinine ratio (mg/g) indicated that the serum creatinine, blood urea, cholesterol, low density lipoprotein, triglyceride, and NRK-52E cells were cultured in vitro, and the cell 2H was treated with iodopol with different iodine concentrations (25,50100140mgI/ml), and the action of iodopol with iodine concentration of 100mgI/ml was fine. Cell 2h, 4h, and 6h. were used to detect the cell apoptosis rate under different iodine concentration and different time by flow Annexin V-FITC/PI double staining. The total protein of each cell was collected and the expression of ZAG protein in each group was detected by Western blot.
Results: of the 10 cases, there were 6 males and 4 females, the average age was 58.40 + 6.50 years. The preoperative urea nitrogen 5.98 + 0.67mmol/L, cholesterol 3.97 + 0.76mmol/L, triglyceride 1.39 + 0.34mmol/L, low density lipoprotein 2.09 + 0.57mmol/L, contrast agent dosage 101 15.24ml, and serum creatinine 63.83 + 9.65umol/L, calculated eGFR as 104.07 + 11.08ml/min 1.73m2. preoperation urine. The level of middle ZAG was 22.85 + 11.10mg/g. after operation 2h, and the level of ZAG in 24h urine was 67.44 + 44.94mg/g, 32.24 + 18.55mg/g respectively. Compared with the level of ZAG in the urine before operation, the ZAG level of 2H urine was higher than that before the operation. The difference was statistically significant (P0.05). There was no correlation with the main indexes. The apoptosis rate of NRK-52E cells cultured in vitro was detected by flow type Annexin V-FITC/PI double staining method with iodopol acting cells with different iodine concentration and iodine concentration of 100mgI/ml. Compared with the control group, the apoptosis rate was iodine concentration (P0.05, n=3). The time dependence (P0.05, n=3) increased, and the expression level of ZAG protein was also detected by Western blot (P0.05, n=3) and time dependence (P0.05, n=3). The linear regression analysis showed that the expression level of ZAG protein was positively correlated with the rate of apoptosis.
Conclusion: 1. Elevated ZAG levels in urine may reflect early renal injury induced by iodinated contrast media.
2, the expression of ZAG protein in CM induced NRK-52E cells is associated with an increased level of apoptosis.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R692.6

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