肿瘤坏死因子α、白细胞介素6与IgA肾病足细胞损伤的相关研究

发布时间：2018-05-19 02:19

本文选题：IgA肾病 + 足细胞　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:近年来,足细胞损伤在Ig A肾病(Ig A nephropathy,Ig AN)中的致病作用受到广泛关注。本研究通过检测Ig AN肾组织肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin-6,IL-6)、Wilms肿瘤蛋白(Wilm’s tumor 1,WT1)的表达变化及尿液中TNF-α、IL-6的水平,并通过收集临床和病理资料,拟探讨TNF-α、IL-6在Ig AN足细胞损伤及疾病发生发展中的作用。方法:选取2015年12月至2016年12月在河北医科大学第二医院住院,行肾穿刺活检术,根据临床和病理资料确诊为Ig AN患者40例。均除外过敏性紫癜、自身免疫性疾病、肝硬化、银屑病、强直性脊柱炎等继发性Ig AN,并除外合并其他肾病,如糖尿病肾病等。患者的病理指标按Ig A肾病的Lee氏分级,并参照Katafuchi等的方法对肾小球、小管间质等进行半定量积分。分别分析各临床指标与不同病理类型及病理改变积分之间的关系。按Lee氏肾组织改变分为I~V级,其中I~III级为一组,IV、V级为另外一组。收集患者一般资料:性别、年龄、血压;临床指标:尿蛋白、血白蛋白、血肌酐、尿酸、β2-微球蛋白、总胆固醇、甘油三酯、空腹血糖、尿渗透压,应用简化的MDRD公式评估肾小球滤过率。河北医科大学第二医院体检中心健康体检者40例作为对照组。河北医科大学第二医院泌尿外科肾脏肿瘤切除术后,远离病灶部位的肾脏组织10例作为正常组织对照。采用酶联免疫吸附法测定尿TNF-α、尿IL-6水平。尿TNF-α、尿IL-6浓度均除以尿肌酐值进行校正。应用免疫组织化学法做TNF-α、IL-6、WT1的肾组织染色,并用图像分析软件进行分析。应用IBM spss21.0统计学软进行统计学分析。结果:1研究对象的一般资料:(1)与对照组相比,Ig AN患者收缩压、舒张压、血肌酐、尿酸、β2-微球蛋白、总胆固醇、甘油三酯明显升高,血白蛋白、e GFR明显降低,差异有统计学意义(P0.05),两组患者性别、年龄、空腹血糖的差异无统计学意义(P0.05)。(2)实验组两组间比较:与I~III级患者相比,Ig AN IV、V级患者收缩压、舒张压、尿蛋白、血肌酐、尿酸、β2-微球蛋白、总胆固醇、甘油三酯明显升高,血白蛋白、e GFR、尿渗透压明显降低,差异有统计学意义(P0.05),两组间性别、年龄、空腹血糖的差异均无统计学意义(P0.05)。2 TNF-α的变化:(1)肾组织中TNF-α的表达:TNF-α主要表达于近端小管上皮细胞胞浆内,其中以萎缩的近端小管最明显,成黄色或棕黄色颗粒。实验组两组比较,I~III级Ig AN患者肾组织中TNF-α的平均光密度为(17.12±5.22),IV、V级Ig AN患者肾组织中TNF-α的平均光密度为(35.15±9.42),差异有统计学意义(P0.05)。(2)Ig AN患者尿TNF-α的水平为(23.18±10.09)ng/mg·Cr,正常对照组尿TNF-α的水平为(11.92±2.53)ng/mg·Cr,二者相比,实验组明显升高,差异有统计学意义(P0.05);实验组两组比较,I~III级Ig AN患者尿TNF-α的水平为(15.70±4.98)ng/mg·Cr,IV、V级Ig AN患者尿TNF-α的水平为(29.95±8.49)ng/mg·Cr,病理类型越重,尿TNF-α的水平越高,差异有统计学意义(P0.05)。3 IL-6的变化:(1)肾组织中IL-6的表达:IL-6主要表达于近端小管上皮细胞胞浆内,其中以萎缩的近端小管最明显,成黄色或棕黄色颗粒。实验组两组比较,I~III级Ig AN患者肾组织中IL-6的平均光密度为(24.25±11.98),IV、V级Ig AN患者肾组织中IL-6的平均光密度为(52.57±20.15),差异有统计学意义(P0.05)。(2)Ig AN患者尿IL-6的水平为(20.45±10.34)pg/mg·Cr,正常对照组尿IL-6的水平为(5.90±2.31)pg/mg·Cr,二者相比,实验组明显升高,差异有统计学意义(P0.05);实验组两组比较,I~III级Ig AN患者尿IL-6的水平为(13.18±5.30)ng/mg·Cr,IV、V级Ig AN患者尿IL-6的水平为(27.01±9.36)ng/mg·Cr,病理类型越重,尿IL-6的水平越高,差异有统计学意义(P0.05)。4肾组织中WT1的表达:WT1主要表达于正常足细胞的细胞核,成黄色或棕黄色颗粒。实验组两组比较,I~III级Ig AN患者肾组织中WT1的平均光密度为(14.15±5.30),IV、V级Ig AN患者肾组织中WT1的平均光密度为(9.44±3.51),差异有统计学意义(P0.05)。5相关性分析结果:(1)肾组织中TNF-α与各指标相关分析:Ig AN患者肾组织中TNF-α的表达与收缩压、舒张压、尿蛋白、血肌酐、尿酸、β2-微球蛋白、总胆固醇、甘油三酯、系膜增生、肾小球硬化度、间质纤维化、肾小管萎缩、IL-6阳性表达率、尿TNF-α、尿IL-6呈正相关,与血白蛋白、e GFR、尿渗透压、WT1阳性表达率呈负相关,与年龄、空腹血糖无明显相关性。(2)尿TNF-α与各指标相关分析:Ig AN患者尿TNF-α水平与收缩压、舒张压、尿蛋白、血肌酐、尿酸、β2-微球蛋白、总胆固醇、系膜增生、肾小球硬化度、间质纤维化、肾小管萎缩、TNF-α阳性表达率、IL-6阳性表达率、尿IL-6呈正相关,与血白蛋白、e GFR、尿渗透压、w T1阳性表达率呈负相关,与年龄、空腹血糖、甘油三酯无明显相关性。(3)肾组织中IL-6与各指标相关分析:Ig AN患者肾组织中IL-6的表达与收缩压、舒张压、尿蛋白、血肌酐、尿酸、β2-微球蛋白、总胆固醇、系膜增生、肾小球硬化度、间质纤维化、肾小管萎缩、TNF-α阳性表达率、尿TNF-α、尿IL-6呈正相关,与血白蛋白、e GFR、尿渗透压、w T1阳性表达率呈负相关,与年龄、空腹血糖、甘油三酯无明显相关性。(4)尿IL-6与各指标相关分析:Ig AN患者尿IL-6水平与收缩压、舒张压、尿蛋白、血肌酐、尿酸、β2-微球蛋白、总胆固醇、系膜增生、肾小球硬化度、间质纤维化、肾小管萎缩、TNF-α阳性表达率、IL-6阳性表达率、尿TNF-α呈正相关,与血白蛋白、e GFR、尿渗透压、w T1阳性表达率呈负相关,与年龄、空腹血糖、甘油三酯无明显相关性。结论:1 TNF-α、IL-6加重Ig AN足细胞损伤,促进疾病进展。2尿液TNF-α、IL-6的水平有望成为评价肾组织局部TNF-α、IL-6代谢的可靠指标,为临床监测Ig AN进展提供一项无创伤的检查方法。
[Abstract]:Objective: in recent years, the pathogenicity of podocyte injury in Ig A nephropathy (Ig A nephropathy, Ig AN) has been widely concerned. This study was conducted by detecting the expression of Ig AN renal tissue tumor necrosis factor alpha (tumor necrosis factor- alpha), interleukin 6, and the changes in the expression of tumor protein 1. NF- alpha, IL-6 level, and by collecting clinical and pathological data, we should explore the role of TNF- alpha and IL-6 in Ig AN foot cell injury and the development of disease. Methods: from December 2015 to December 2016, the second hospital of Hebei Medical University was hospitalized, and the renal biopsy was performed. According to the clinical and pathological data, 40 cases of Ig AN patients were confirmed. All of them were excluded. Anaphylactoid purpura, autoimmune disease, cirrhosis, psoriasis, ankylosing spondylitis, and other secondary Ig AN, except for other nephropathy, such as diabetic nephropathy. The pathological indexes of the patients are classified according to Lee's grade of Ig A nephropathy, and the glomeruli, tubulointerstitium, etc. are integrated with the methods of Katafuchi and so on. The relationship between the standard and the score of pathological changes and pathological changes was divided into I~V grade according to Lee's renal tissue changes, of which I~III was a group, IV, and V was another group. The general data of the patients were collected: sex, age, blood pressure, and clinical indicators: urinary protein, serum albumin, serum creatinine, uric acid, beta 2- microglobulin, total cholesterol, triglyceride, fasting blood glucose The urine osmotic pressure was used to evaluate the glomerular filtration rate by the simplified MDRD formula. 40 cases of healthy physical examination at the physical examination center of the second hospital of Hebei Medical University were used as the control group. After the kidney tumor resection in the Department of Urology of the second hospital of Hebei Medical University, 10 cases of renal tissue far away from the lesion were used as normal tissue control. The enzyme linked immunosorbent assay was used. Urine TNF- alpha, urine IL-6 level, urine TNF- alpha, urine IL-6 concentration divided by urine creatinine value for correction. Immunohistochemical staining was used to do TNF- alpha, IL-6, WT1 renal tissue staining, and analyzed using image analysis software. IBM spss21.0 statistics soft for statistical analysis. 1 general data: (1) compared with the control group, I G AN systolic pressure, diastolic pressure, serum creatinine, uric acid, beta 2- microglobulin, total cholesterol, triglyceride significantly increased, serum albumin, e GFR significantly decreased, the difference was statistically significant (P0.05). The differences in sex, age, and fasting blood glucose in the two groups were not statistically significant (P0.05). (2) comparison between the two groups in the experimental group: Ig AN IV compared with the I~III level patients. The systolic pressure, diastolic pressure, urine protein, serum creatinine, uric acid, beta 2- microglobulin, total cholesterol, triglyceride, serum albumin, e GFR, urinary osmotic pressure decreased significantly (P0.05). There was no significant difference in sex, age, and fasting blood glucose between the two groups (P0.05) the changes of.2 TNF- A: (1) TNF- in the renal tissue. The expression of TNF- alpha was mainly expressed in the cytoplasm of proximal tubular epithelial cells. The atrophy proximal tubules were the most obvious, yellow or brown granules. The average optical density of TNF- alpha in the renal tissue of the I~III grade Ig AN patients was (17.12 + 5.22), IV, and the average optical density of TNF- a in the renal tissue of V grade Ig AN was (35.15 + 9.42). The difference was statistically significant (P0.05). (2) the level of urinary TNF- alpha in patients with Ig AN was (23.18 + 10.09) ng/mg. Cr, and the level of urine TNF- a in the normal control group was (11.92 + 2.53) ng/mg. Cr, the experimental group was significantly higher than the two, and the difference was statistically significant (P0.05). The level of two groups in the test group was (15.70 + 4.98). The level of urine TNF- alpha in patients with Cr, IV, V Ig AN was (29.95 + 8.49) ng/mg. Cr, the heavier the pathological type, the higher the level of urinary TNF- alpha, the difference was statistically significant (P0.05).3 IL-6: (1) the expression in the renal tissue was mainly expressed in the proximal tubule cell cytoplasm, and the atrophied proximal tubules were most obvious, yellow or brown. The average optical density of IL-6 in renal tissue of I~III class Ig AN patients was (24.25 + 11.98), and the average optical density of IL-6 in renal tissue of IV and V grade Ig AN patients was (52.57 + 20.15), the difference was statistically significant (P0.05). (2) the level of urinary tract of Ig AN patients was (20.45 + 10.34), and the level of urine samples in normal control group was 5.. 90 + 2.31) pg/mg. Cr, compared with the two, the experimental group was significantly higher, and the difference was statistically significant (P0.05). The level of urinary IL-6 in the two groups of the experimental group was (13.18 + 5.30) ng/mg. Cr, IV, V class Ig. The higher the pathological type, the higher the level of urinary tract, the difference was statistically significant. 05) the expression of WT1 in.4 renal tissue: WT1 was mainly expressed in the nucleus of normal poddal and yellow or brown yellow granules. The average optical density of WT1 in the renal tissue of the I~III class Ig AN patients was (14.15 + 5.30), IV and V Ig AN were (9.44 + 3.51), and the difference was statistically significant. The results of the correlation analysis: (1) the correlation analysis of TNF- a in renal tissue: the expression of TNF- alpha in renal tissue of patients with Ig AN and systolic pressure, diastolic pressure, urinary protein, serum creatinine, uric acid, beta 2- microglobulin, total cholesterol, triglyceride, mesangial hyperplasia, glomerulosclerosis, interstitial fibrosis, renal tubule atrophy, IL-6 positive expression, urinary TNF- a, urinary IL-6 Positive correlation was negative correlation with serum albumin, e GFR, urine osmotic pressure and WT1 positive expression, and no significant correlation with age and fasting blood glucose. (2) urinary TNF- alpha was associated with various indexes: the level of urinary TNF- A and systolic pressure, diastolic pressure, urinary protein, serum creatinine, uric acid, beta 2- microglobulin, total cholesterol, mesangial hyperplasia, glomerulosclerosis Interstitial fibrosis, renal tubule atrophy, positive expression of TNF- alpha, positive rate of IL-6, positive correlation of urinary IL-6, negative correlation with serum albumin, e GFR, urinary osmotic pressure, positive expression of W T1, and no significant correlation with age, fasting blood glucose and triglyceride. (3) the correlation of IL-6 in renal tissue with each index: the IL-6 expression and collection in the renal tissue of Ig AN patients Contraction pressure, diastolic pressure, urine protein, serum creatinine, uric acid, beta 2- microglobulin, total cholesterol, mesangial hyperplasia, glomerulosclerosis, interstitial fibrosis, renal tubule atrophy, TNF- alpha positive expression, urinary TNF- alpha, IL-6 positive correlation, negative correlation with serum albumin, e GFR, urinary osmotic pressure, w T1 positive expression, and age, fasting glycemia, triglyceride free Significant correlation. (4) urine IL-6 and the correlation analysis: IL-6 level and systolic pressure, diastolic pressure, diastolic pressure, urinary protein, creatinine, uric acid, beta 2- microglobulin, total cholesterol, mesangial hyperplasia, glomerulosclerosis, interstitial fibrosis, renal tubule atrophy, TNF- alpha positive expression rate, IL-6 positive expression rate, positive correlation of urinary TNF- a, and serum albumin, e G FR, urinary osmotic pressure and w T1 positive expression have negative correlation with age, fasting blood glucose and triglyceride. Conclusion: 1 TNF- alpha, IL-6 aggravates Ig AN foot cell injury and promotes the progression of.2 urine TNF- alpha. The level of IL-6 is expected to be a reliable index for evaluating the local TNF- alpha and IL-6 metabolism. A noninvasive method of examination.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692.31

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