氧化應(yīng)激在造影劑腎病中的作用及干預(yù)研究
發(fā)布時(shí)間：2019-05-22 00:08
【摘要】：背景和目的 隨著現(xiàn)代醫(yī)學(xué)影像診斷和介入治療的快速發(fā)展，造影劑腎病(CIN)的發(fā)病率呈上升趨勢(shì)，其發(fā)病機(jī)制、干預(yù)措施也受到更多的重視。老年糖尿病人群中CIN的發(fā)病機(jī)制尚未完全明確，氧化應(yīng)激以及信號(hào)傳導(dǎo)是目前研究的熱點(diǎn)。我們通過制作老年糖尿病大鼠模型，觀察造影劑對(duì)腎功能和腎組織一氧化氮(NO)、一氧化氮合成酶(NOS)及谷胱甘肽(GSH)的影響，探討了NO、NOS及GSH在CIN發(fā)病機(jī)制中的作用。繼而構(gòu)建表達(dá)了兩種NOS的嵌合體酶，研究了嵌合體酶的類型和結(jié)構(gòu)對(duì)NO合成活性的影響。臨床工作中對(duì)于老年患者，，通過肌酐或者由肌酐計(jì)算得出的腎小球?yàn)V過率可能無法準(zhǔn)確反映腎功能的變化，從而延誤CIN的診斷。本論文通過對(duì)白細(xì)胞介素18和胱抑素C在老年糖尿病患者中的表達(dá)研究,尋找早期、敏感、特異預(yù)測(cè)CIN的生物學(xué)指標(biāo)。考慮到氧化應(yīng)激在CIN發(fā)病機(jī)理中的作用，抗氧化劑對(duì)于高危人群預(yù)防CIN的發(fā)生可能有一定的效果。GSH作為復(fù)雜的抗氧化網(wǎng)絡(luò)的中心，其功能是通過緩沖活性氧、調(diào)節(jié)NO循環(huán)和改善控制著生長、發(fā)展和防御的細(xì)胞氧化還原信號(hào)傳遞系統(tǒng)來體現(xiàn)的。評(píng)價(jià)還原型谷胱甘肽聯(lián)合生理鹽水水化對(duì)老年糖尿病患者CIN干預(yù)效果。由于血液中的存在著大量GSH，本論文還研究了GSH與血紅蛋白的結(jié)合過程的機(jī)理，進(jìn)一步了解GSH發(fā)揮作用的機(jī)制。 本論文的研究內(nèi)容主要包括三個(gè)部分。 第一部分：低滲造影劑對(duì)老年糖尿病大鼠腎毒性及一氧化氮合成酶對(duì)一氧化氮合成機(jī)理影響的研究 一、材料與方法方法是通過建立老年糖尿病大鼠模型，觀察尾靜脈注射歐乃派克350(10ml/kg)后，大鼠血肌酐、尿素氮以及腎組織勻漿NO、NOS和GSH的含量。構(gòu)建表達(dá)了兩種NOS的一氧化氮合成酶，獲取了一氧化氮合成酶的光譜，研究了一氧化氮合成酶的NO合成活性，測(cè)試了對(duì)鐵氰化物、細(xì)胞色素C還原速率、黃素、NADPH氧化還原速率的影響。 二、結(jié)果 結(jié)果是老年糖尿病大鼠血肌酐、尿素氮與正常對(duì)照組比較均升高(P 0.05)，腎組織勻漿NO、NOS、GSH與正常對(duì)照組比較均下降。注射造影劑后，血肌酐與正常對(duì)照組比較明顯升高(P 0.01)，腎組織勻漿NO和NOS與糖尿病對(duì)照組比較下降(P 0.05)。nNOSeFNR一氧化氮合成酶表現(xiàn)出較低的血紅素還原速率。 第二部分：白細(xì)胞介素18、胱抑素C對(duì)老年糖尿病患者造影劑腎病的預(yù)測(cè)作用研究 一、材料與方法選擇2011年1月到2011年10月在我院接受增強(qiáng)CT檢查的年齡大于60歲、糖尿病史1年以上、無明顯腎功能異常的患者共30例，患者均采用低滲非離子型造影劑。ELISA法測(cè)定造影前和造影后24小時(shí)、48小時(shí)IL-18以及血尿素氮(BUN)、血肌酐(SCr)、CysC。 二、結(jié)果 觀察對(duì)象均無造影劑腎病的發(fā)生，造影后血IL-18與造影前比較無統(tǒng)計(jì)學(xué)意義(P0.05)，造影后24小時(shí)尿IL-18、血胱抑素C升高明顯，與造影前比較差異有統(tǒng)計(jì)學(xué)意義(P 0.05)，48小時(shí)后SCr升高明顯(P 0.05)。 第三部分：還原型谷胱甘肽對(duì)老年糖尿病患者造影劑腎病影響及與血紅蛋白結(jié)合過程的研究 一、材料與方法 將準(zhǔn)備采用低滲造影劑進(jìn)行增強(qiáng)CT掃描的96位糖尿病患者(年齡≥60歲)隨機(jī)分為兩組。一組靜脈注射還原型谷胱甘肽(于掃描前靜脈注射溶于100ml生理鹽水中的1800mg還原型谷胱甘肽，掃描后同等劑量注射兩天)，同時(shí)以1ml/kg BW/h的速度靜脈注射生理鹽水(GSH組，n=48)；另一組僅以相同速度靜脈注射生理鹽水(對(duì)照組，n=48)。SCr、BUN、CysC和尿IL-18分別于造影前和造影后48小時(shí)測(cè)量。肌酐高于基線0.5mg/dL或者比肌酐基線值增長25%即被確認(rèn)為CIN。利用多種光譜法研究了還原型谷胱甘肽與Hb的結(jié)合過程，確證了GSH對(duì)血紅蛋白的猝滅機(jī)制。根據(jù)所得到的結(jié)合常數(shù)等參數(shù)推斷出GSH與血紅蛋白之間的相互作用力的類型。由F ster能量轉(zhuǎn)移理論可以得到GSH與血紅蛋白的結(jié)合距離。利用多種分子光譜法研究了GSH的存在對(duì)血紅蛋白構(gòu)象的影響。 二、結(jié)果 GSH組和對(duì)照組在基線特征和基線腎功能方面沒有顯著性差異。GSH組CIN的發(fā)病率稍低于對(duì)照組(2.08%vs.8.33%, P=0.364)。增強(qiáng)CT掃描后兩組的SCr、BUN、CysC、尿IL-18均升高，估測(cè)腎小球?yàn)V過率(eGFR)均降低。造影后兩組SCr(P=0.001)，eGFR(P=0.002), BUN(P 0.001)，CysC(P=0.001)和尿IL-18(P 0.001)有顯著性差異。GSH濃度的增加導(dǎo)致Hb的熒光強(qiáng)度逐漸降低。依據(jù)熱力學(xué)原理求得一系列熱力學(xué)參數(shù)，從表中看出, ΔH O、ΔS 0、ΔG O,說明GSH與Hb作用過程是一個(gè)焓變減小、熵減、吉布斯自由能降低的自發(fā)過程，可推斷出GSH與Hb之間的作用力是以氫鍵及范德華力結(jié)合。依據(jù)峰位置的紅移或藍(lán)移來說明氨基酸周圍微環(huán)境的變化，最大發(fā)射波長發(fā)生了明顯的紅移現(xiàn)象，表明GSH改變了Hb中色氨酸殘基所處的微環(huán)境，極性增強(qiáng)。 結(jié)論 1.低滲造影劑可以引起老年糖尿病大鼠腎損傷，NOS、NO的代謝以及信號(hào)傳遞可能與其有關(guān)。 2.一氧化氮合成酶的種類和結(jié)構(gòu)性質(zhì)對(duì)一氧化氮的合成過程有顯著影響。 3.尿IL-18和Cys C是可以早期反映腎損傷的敏感指標(biāo)，對(duì)CIN的早期診斷及防治有重要作用。 4.還原型谷胱甘肽加生理鹽水水化可能通過調(diào)節(jié)氧化和抗氧化系統(tǒng)預(yù)防造影劑腎病的發(fā)生。 5.確定了還原型谷胱甘肽與Hb的結(jié)合過程及相互作用類型。
[Abstract]:Background and Purpose With the rapid development of modern medical image diagnosis and interventional therapy, the incidence of contrast-contrast nephropathy (CIN) is on the rise. The pathogenesis of CIN in the elderly diabetic population is not well defined, oxidative stress and signal transduction are the heat of the present study. Objective: To study the effect of contrast agent on nitric oxide (NO), nitric oxide synthase (NOS) and glutathione (GSH) in renal and renal tissues by making a model of old diabetic rats, and to study the role of NO, NOS and GSH in the pathogenesis of CIN. The type and structure of the chimera enzyme and the effect of the structure on the activity of NO synthesis were studied. In response to a change in renal function that may not be accurately reflected in the clinical work for elderly patients, the rate of glomerular filtration through the muscle or by the muscle tone may not accurately reflect changes in renal function, thereby delaying the diagnosis of the CIN In this paper, the expression of interleukin-18 and cystatin C in elderly patients with diabetes is studied, and the biological means of early, sensitive and specific prediction of CIN are found. The antioxidant has a certain effect on the prevention of CIN in high-risk people, taking into account the role of oxidative stress in the pathogenesis of CIN. The function of GSH as the center of the complex antioxidant network is to reflect the active oxygen, regulate the NO circulation and improve the control of the growth, development and defense of the cell oxidation-reduction signal delivery system Evaluation of the effect of reduced glutathione combined with normal saline on the intervention of CIN in elderly patients with diabetes The present paper also studies the mechanism of the binding process of GSH and hemoglobin in the blood, and further studies the mechanism of GSH. The research content of this paper mainly includes three parts: The first part: hypotonic contrast agent on the renal toxicity and nitric oxide synthase in the aged diabetic rats One, the material and the method of the study is to establish the model of the old diabetic rats, and observe the tail vein injection of Opike 350 (10 ml/ kg). The serum levels of NO, NOS and NO, NOS in the rat's blood muscle, the urine and the kidney tissue were detected. And the activity of NO synthesis of the nitric oxide synthase was studied, and the rate of reduction of iron cyanide and cytochrome C, and the oxidation of the emodin and NADPH were tested. reduction rate The results were as follows: The results showed that the blood myostatin and urea nitrogen in the old diabetic rats were higher than those in the normal control group (P 0.05), and the renal tissue homogenate NO, NOS, and GSH. Compared with the normal control group, the control group of the normal control group decreased significantly (P 0.01), the renal tissue homogenate NO and the NOS and the diabetes control group. Comparative decrease (P 0.05). nNOSeFNR nitric oxide synthase expression lower heme reduction rate, second part: interleukin-18, cystatin C, A study on the predictive effect of contrast-contrast nephropathy in patients was conducted in our hospital from January 2011 to October 2011 in our hospital for enhanced CT examination. A total of 30 patients with a age of more than 60 years, a history of diabetes more than 1 year, and no obvious renal function Low-permeability non-ionic contrast agents were used in the patients. The results of the ELISA method were 24 hours,48 hours IL-18 and blood urea nitrogen (BUN) before and after contrast. ), the blood muscle The results of SCr and CysC.2 showed no contrast-related nephropathy, and there was no significant difference in blood IL-18 after contrast (P0.05). IL-18 and cystatin C in 4-hour urine were significantly higher than those before contrast (P 0.05). SCr increased significantly after 48 hours (P 0.05). Part 3: Reduced glutathione for old-age diabetes Patient contrast nephropathy The study I, materials and methods of the impact and integration with the hemoglobin will be prepared to use hypotonic contrast agents for enhanced CT The scanned 96-bit diabetic patients (aged 60 years) were randomly divided into two groups. A group of intravenous reduced glutathione (1800 mg of reduced glutathione in 100 ml of saline prior to the scan, two days after the same dose of injection), and at the same time at 1 ml/ kg B The velocity of W/ h was injected intravenously (GSH group, n = 48); the other The group was given normal saline at the same speed (control group, n = 48). SCr, BUN, Cy SC and urine IL-18 were measured at 48 hours prior to and after contrast, respectively, and the muscle strength was higher than the baseline of 0.5 m. A 25% increase in g/ dL or greater than the baseline value of the muscle was confirmed to be CIN. Reduced glutathione was studied using a variety of spectrometry In the process of binding to Hb, the quenching mechanism of GSH on the hemoglobin was confirmed. The type of interaction force between the GSH and the hemoglobin is deduced by the parameters such as F st. The combined distance of GSH and hemoglobin can be obtained by the theory of energy transfer. molecular light The effect of the presence of GSH on the conformation of hemoglobin was studied in this paper. There was no significant difference between the baseline characteristics and the baseline renal function in the GSH group and the control group. Compared with the control group (2.08% vs. 8.33%, P = 0.364), SCr, BUN, and Cys in the two groups after enhanced CT scan C and urinary IL-18 both increased, and the estimated glomerular filtration rate (eGFR) was decreased. Two groups of SCr (P = 0.001), eGFR (P = 0.002), BUN (P 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), P = 0.001) and urine IL-18 (P 0.001 There is a significant difference. The increase of the concentration of GSH results in a gradual decrease of the fluorescence intensity of Hb. A series of thermodynamic parameters are obtained from the thermodynamic principle. b The process of action is a spontaneous over-reduction, entropy reduction, Gibbs free energy reduction, It can be concluded that the force between the GSH and the Hb is combined with the Van der Waals force. The change of the micro-environment around the amino acid is explained according to the red shift or the blue shift of the peak position, and the maximum emission wavelength has obvious red shift. like, Conclusion 1. The low-permeability contrast agent can be cited. The metabolism of NOS and NO and the transmission of signal may be related to the renal injury, NOS and NO in the old diabetic rats. 2. The type and structure of nitric oxide synthase have a significant effect on the synthesis of nitric oxide. Cys C is a sensitive index that can reflect the renal injury in an early stage, and plays an important role in the early diagnosis and control of CIN. . The hydration of reduced glutathione and physiological saline may be regulated by the regulation of oxidation and anti-oxidation.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692
本文編號(hào)：2482519
[Abstract]:Background and Purpose With the rapid development of modern medical image diagnosis and interventional therapy, the incidence of contrast-contrast nephropathy (CIN) is on the rise. The pathogenesis of CIN in the elderly diabetic population is not well defined, oxidative stress and signal transduction are the heat of the present study. Objective: To study the effect of contrast agent on nitric oxide (NO), nitric oxide synthase (NOS) and glutathione (GSH) in renal and renal tissues by making a model of old diabetic rats, and to study the role of NO, NOS and GSH in the pathogenesis of CIN. The type and structure of the chimera enzyme and the effect of the structure on the activity of NO synthesis were studied. In response to a change in renal function that may not be accurately reflected in the clinical work for elderly patients, the rate of glomerular filtration through the muscle or by the muscle tone may not accurately reflect changes in renal function, thereby delaying the diagnosis of the CIN In this paper, the expression of interleukin-18 and cystatin C in elderly patients with diabetes is studied, and the biological means of early, sensitive and specific prediction of CIN are found. The antioxidant has a certain effect on the prevention of CIN in high-risk people, taking into account the role of oxidative stress in the pathogenesis of CIN. The function of GSH as the center of the complex antioxidant network is to reflect the active oxygen, regulate the NO circulation and improve the control of the growth, development and defense of the cell oxidation-reduction signal delivery system Evaluation of the effect of reduced glutathione combined with normal saline on the intervention of CIN in elderly patients with diabetes The present paper also studies the mechanism of the binding process of GSH and hemoglobin in the blood, and further studies the mechanism of GSH. The research content of this paper mainly includes three parts: The first part: hypotonic contrast agent on the renal toxicity and nitric oxide synthase in the aged diabetic rats One, the material and the method of the study is to establish the model of the old diabetic rats, and observe the tail vein injection of Opike 350 (10 ml/ kg). The serum levels of NO, NOS and NO, NOS in the rat's blood muscle, the urine and the kidney tissue were detected. And the activity of NO synthesis of the nitric oxide synthase was studied, and the rate of reduction of iron cyanide and cytochrome C, and the oxidation of the emodin and NADPH were tested. reduction rate The results were as follows: The results showed that the blood myostatin and urea nitrogen in the old diabetic rats were higher than those in the normal control group (P 0.05), and the renal tissue homogenate NO, NOS, and GSH. Compared with the normal control group, the control group of the normal control group decreased significantly (P 0.01), the renal tissue homogenate NO and the NOS and the diabetes control group. Comparative decrease (P 0.05). nNOSeFNR nitric oxide synthase expression lower heme reduction rate, second part: interleukin-18, cystatin C, A study on the predictive effect of contrast-contrast nephropathy in patients was conducted in our hospital from January 2011 to October 2011 in our hospital for enhanced CT examination. A total of 30 patients with a age of more than 60 years, a history of diabetes more than 1 year, and no obvious renal function Low-permeability non-ionic contrast agents were used in the patients. The results of the ELISA method were 24 hours,48 hours IL-18 and blood urea nitrogen (BUN) before and after contrast. ), the blood muscle The results of SCr and CysC.2 showed no contrast-related nephropathy, and there was no significant difference in blood IL-18 after contrast (P0.05). IL-18 and cystatin C in 4-hour urine were significantly higher than those before contrast (P 0.05). SCr increased significantly after 48 hours (P 0.05). Part 3: Reduced glutathione for old-age diabetes Patient contrast nephropathy The study I, materials and methods of the impact and integration with the hemoglobin will be prepared to use hypotonic contrast agents for enhanced CT The scanned 96-bit diabetic patients (aged 60 years) were randomly divided into two groups. A group of intravenous reduced glutathione (1800 mg of reduced glutathione in 100 ml of saline prior to the scan, two days after the same dose of injection), and at the same time at 1 ml/ kg B The velocity of W/ h was injected intravenously (GSH group, n = 48); the other The group was given normal saline at the same speed (control group, n = 48). SCr, BUN, Cy SC and urine IL-18 were measured at 48 hours prior to and after contrast, respectively, and the muscle strength was higher than the baseline of 0.5 m. A 25% increase in g/ dL or greater than the baseline value of the muscle was confirmed to be CIN. Reduced glutathione was studied using a variety of spectrometry In the process of binding to Hb, the quenching mechanism of GSH on the hemoglobin was confirmed. The type of interaction force between the GSH and the hemoglobin is deduced by the parameters such as F st. The combined distance of GSH and hemoglobin can be obtained by the theory of energy transfer. molecular light The effect of the presence of GSH on the conformation of hemoglobin was studied in this paper. There was no significant difference between the baseline characteristics and the baseline renal function in the GSH group and the control group. Compared with the control group (2.08% vs. 8.33%, P = 0.364), SCr, BUN, and Cys in the two groups after enhanced CT scan C and urinary IL-18 both increased, and the estimated glomerular filtration rate (eGFR) was decreased. Two groups of SCr (P = 0.001), eGFR (P = 0.002), BUN (P 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), CysC (P = 0.001), P = 0.001) and urine IL-18 (P 0.001 There is a significant difference. The increase of the concentration of GSH results in a gradual decrease of the fluorescence intensity of Hb. A series of thermodynamic parameters are obtained from the thermodynamic principle. b The process of action is a spontaneous over-reduction, entropy reduction, Gibbs free energy reduction, It can be concluded that the force between the GSH and the Hb is combined with the Van der Waals force. The change of the micro-environment around the amino acid is explained according to the red shift or the blue shift of the peak position, and the maximum emission wavelength has obvious red shift. like, Conclusion 1. The low-permeability contrast agent can be cited. The metabolism of NOS and NO and the transmission of signal may be related to the renal injury, NOS and NO in the old diabetic rats. 2. The type and structure of nitric oxide synthase have a significant effect on the synthesis of nitric oxide. Cys C is a sensitive index that can reflect the renal injury in an early stage, and plays an important role in the early diagnosis and control of CIN. . The hydration of reduced glutathione and physiological saline may be regulated by the regulation of oxidation and anti-oxidation.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 Nilesh Lodhia;Michael Kader;Thalia Mayes;Parvez Mantry;Benedict Maliakkal;;Risk of contrast-induced nephropathy in hospitalized patients with cirrhosis[J];World Journal of Gastroenterology;2009年12期
2 王忠良;劉敏;張義勤;;丹紅注射液對(duì)PCI術(shù)后造影劑腎損害的預(yù)防作用[J];中國中西醫(yī)結(jié)合雜志;2011年12期
本文編號(hào)：2482519
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