他克莫司对激素耐药型肾病综合征患儿MDR1及其P-gp170表达的影响及意义

发布时间：2017-12-27 10:10

本文关键词：他克莫司对激素耐药型肾病综合征患儿MDR1及其P-gp170表达的影响及意义　出处：《苏州大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的:检测原发性肾病综合征(INS)患儿不同时间点外周血单个核细胞(PBMC)多药耐药基因(MDR1)及其糖蛋白170(P-gp170)的表达情况,探讨激素耐药型肾病综合征(SRNS)患儿加用他克莫司(TAC)治疗后其MDR1及其P-gp170表达的变化和临床意义。方法:选择本院住院的原发性肾病综合征(INS)患儿36例为研究对象,根据激素治疗效果分成两组:激素敏感型肾病综合征(SSNS)组20例和激素耐药型肾病综合征(SRNS)组16例;20名健康体检儿童作为正常对照组(control)。分四个时间点采集外周血标本:(1)发病初未用糖皮质激素(GC)时;(2)足量GC治疗4周后;(3)SSNS组的GC减量4周后或SRNS组的加用TAC治疗4周后;(4)SRNS组的加用TAC治疗8周后。进行如下实验:①实时定量逆转录聚合酶链式反应(RT-PCR)检测SRNS和SSNS不同时间点PBMC MDR1m RNA的表达。②流式细胞仪(FCM)检测SRNS和SSNS不同时间点PBMC中淋巴细胞(Lymphocytes)上P-gp170的表达百分数。③16例SRNS患儿均在激素治疗4周后行肾穿刺活检术,10例正常肾组织为正常肾组织对照组,用免疫组织化学方法(IHC)检测肾组织中P-gp170的表达。结果:(1)INS患儿PBMC MDR1m RNA与Lymphocytes P-gp170的表达呈高度正相关(r=0.854,P0.01)。(2)GC治疗前,INS患儿PBMC MDR1m RNA与Lymphocytes P-gp170表达量均高于Control组,差异具有统计学意义(P0.05)。(3)足量GC治疗4周后,SSNS组和SRNS组患儿PBMC MDR1m RNA及Lymphocytes P-gp170的表达均较治疗前明显升高,差异均具统计学意义(P0.01)。(4)GC减量4周后,SSNS患儿MDR1m RNA及Lymphocytes P-gp170的表达较前明显回落,差异均具统计学意义(P0.01),而与GC治疗前无明显差异性(P0.05)。(5)加用TAC治疗4、8周后,SRNS组患儿PBMC MDR1m RNA及Lymphocytes P-gp170的表达均较前明显回落(P0.01),而加用TAC治疗4周后的SRNS患儿两指标的表达量明显高于GC减量4周后的SSNS患儿(P0.01)。(6)在SRNS组中,TAC治疗8周后PBMC MDR1m RNA及Lymphocytes P-gp170的表达量明显低于4周后(P0.01),且与GC治疗前相比,无明显统计学差异(P0.05)。(7)SRNS患儿与正常肾组织对照组儿童肾组织中P-gp170主要表达于肾小管及肾间质,SRNS组肾组织近曲小管和远曲小管的P-gp170阳性率表达均明显高于正常肾组织对照组(P0.01),SRNS组内近曲小管P-gp170阳性率表达明显高于远曲小管(P0.01)。(8)足量GC治疗4周后,SRNS患儿Lymphocytes P-gp170的表达量与肾组织近曲小管及远曲小管P-gp170的表达量均呈正相关(r=0.547,P0.05;r=0.541,P0.05)。(9)GC治疗前,SRNS组患儿PBMC P-gp170的表达量与24小时尿蛋白定量正相关(r=0.712,P0.01)。结论:(1)原发性肾病综合征患儿PBMC MDR1m RNA与Lymphocytes P-gp170的表达呈高度正相关,即MDR1m RNA表达量越高,其蛋白产物P-gp170表达量越高;(2)激素耐药型肾病综合征(SRNS)患儿PBMC MDR1m RNA及其蛋白产物P-gp170的高表达可能参与了激素耐药的发生和发展;(3)SRNS患儿的肾组织P-gp170的高表达可能与激素耐药有关。(4)他克莫司可在一定程度上减少SRNS患儿PBMC MDR1m RNA与P-gp170的表达量,可能为逆转肾病综合征患儿激素耐药的机制之一。
[Abstract]:Objective: to detect the primary nephrotic syndrome (INS) patients at different time points of peripheral blood mononuclear cells (PBMC) multidrug resistance gene (MDR1) and glycoprotein 170 (P-gp170) expression, to explore the steroid resistant nephrotic syndrome (SRNS) patients with tacrolimus (TAC) and the clinical significance of changes of expression the MDR1 and P-gp170 after treatment. Methods: our hospital with primary nephrotic syndrome (INS) patients with 36 cases as the research object, according to the effect of hormone therapy were divided into two groups: steroid sensitive nephrotic syndrome (SSNS) group and 20 cases of steroid resistant nephrotic syndrome (SRNS) group of 16 cases; 20 healthy children as the normal control group (control). The peripheral blood samples were collected at four time points: (1) when the onset was not used Glucocorticoid (GC), (2) the full dose of GC was treated for 4 weeks; (3) the GC reduction in group SSNS was 4 weeks after treatment, or the SRNS group was treated with TAC for 4 weeks; (4) SRNS group was treated with TAC for 8 weeks. The following experiments were carried out: (1) real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of PBMC MDR1m RNA at different time points of SSNS. (2) flow cytometry (FCM) was used to detect the percentage of P-gp170 expression on the lymphocyte (Lymphocytes) in PBMC at different time points in SRNS and SSNS. (3) 16 children with SRNS underwent renal biopsy after 4 weeks of glucocorticoid treatment, and 10 normal kidney tissues were normal renal tissue control group. The expression of P-gp170 in renal tissue was detected by immunohistochemical method (IHC). Results: (1) the expression of PBMC MDR1m RNA in children with INS was highly correlated with the expression of Lymphocytes P-gp170 (r=0.854, P0.01). (2) before GC treatment, the expression of PBMC MDR1m RNA and Lymphocytes P-gp170 in children with INS was higher than that in Control group, and the difference was statistically significant (P0.05). (3) after 4 weeks of adequate GC treatment, the expressions of PBMC MDR1m RNA and Lymphocytes P-gp170 in SSNS group and SRNS group were significantly increased compared with those before treatment, and the differences were statistically significant (P0.01). (4) after 4 weeks of GC reduction, the expression of MDR1m RNA and Lymphocytes P-gp170 in SSNS children was significantly lower than before. The difference was statistically significant (P0.01), but there was no significant difference between GC and GC (P0.05). (5) after adding TAC for 4 or 8 weeks, the expression of PBMC MDR1m RNA and Lymphocytes P-gp170 in group SRNS was significantly lower than before (P0.01), while the expression of two index in SRNS children treated with TAC for 4 weeks was significantly higher than that in 4 children (4). (6) in group SRNS, after 8 weeks of TAC treatment, the expression of PBMC MDR1m RNA and Lymphocytes P-gp170 was significantly lower than that after 4 weeks (P0.01), and there was no significant difference compared with that before GC treatment (P0.05). (7) SRNS patients and normal renal tissues of children in control group P-gp170 in renal tissue was mainly expressed in renal tubular and interstitial, the positive rate of P-gp170 in renal tissue of SRNS group, the distal and proximal convoluted tubules expression were significantly higher than normal renal tissue of control group (P0.01), was significantly higher than the positive rate of proximal convoluted tubules tubular P-gp170 in the SRNS group (P0.01). (8) after 4 weeks of adequate GC treatment, the expression of Lymphocytes P-gp170 in SRNS children was positively correlated with the expression of P-gp170 in renal proximal tubules and distal tubules (r=0.547, P0.05, r=0.541, P0.05). (9) before GC treatment, the expression of PBMC P-gp170 in the children of group SRNS was positively correlated with the 24 hour urine protein (r=0.712, P0.01). Conclusion: (1) primary nephrotic syndrome in children with PBMC MDR1m RNA and Lymphocytes P-gp170 were highly correlated, MDR1m expression level of RNA is higher, the protein expression of P-gp170 is higher; (2) steroid resistant nephrotic syndrome (SRNS) patients with high expression of PBMC MDR1m RNA and its protein product P-gp170 may be involved in the occurrence and development of hormone resistance; (3) the high expression of SRNS in renal tissue of P-gp170 may be related to hormone resistance. (4) tacrolimus can reduce the expression of PBMC MDR1m RNA and P-gp170 to some extent in children with SRNS, which may be one of the mechanisms of reversing steroid resistance in children with nephrotic syndrome.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R726.9

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