發(fā)布時(shí)間：2018-08-03 18:26

【摘要】：目的檢測P-糖蛋白170(P-gp170)及熱休克蛋白90(HSP90)在原發(fā)性腎病綜合征(PNS)患兒不同時(shí)間點(diǎn)外周血淋巴細(xì)胞及腎組織中的表達(dá)情況，探討P-gp170與HSP90介導(dǎo)糖皮質(zhì)激素(GC)耐藥的可能機(jī)制及其臨床意義。 方法選擇在蘇州大學(xué)附屬兒童醫(yī)院住院的PNS患兒20例，按照GC對患兒的療效分成兩組：激素耐藥型腎病綜合征(SRNS)和激素敏感型腎病綜合征(SSNS),各10名，10名健康兒童作為正常對照組，分兩個(gè)時(shí)間點(diǎn)采集外周血標(biāo)本：（1）發(fā)病初未用GC時(shí)；（2）足量GC治療4周后。進(jìn)行如下實(shí)驗(yàn)：①流式細(xì)胞術(shù)(FCM)檢測PNS患兒及正常對照組在不同時(shí)間點(diǎn)外周血淋巴細(xì)胞中P-gp170及HSP90的表達(dá)。②10例SRNS患兒均在激素治療4周后行腎穿刺活檢術(shù)，5例正常腎組織為對照，用免疫組織化學(xué)方法檢測腎組織中P-gp170及HSP90的表達(dá)。 結(jié)果(1)SSNS組、SRNS組和正常對照組外周血淋巴細(xì)胞中均有P-gp170及HSP90的表達(dá)；(2)激素治療前SSNS組、SRNS組外周血淋巴細(xì)胞中的P-gp170及HSP90高于正常對照組(P0.05)；激素治療4周后SSNS組及SRNS組外周血淋巴細(xì)胞中的P-gp170明顯高于治療前(P0.05)，HSP90較前下降(P0.05)；(3)激素治療前SRNS組外周血淋巴細(xì)胞中P-gp170明顯高于SSNS組及正常對照組，，治療后SRNS組外周血淋巴細(xì)胞中P-gp170較治療前升高(P0.05)；激素治療前SRNS組外周血淋巴細(xì)胞中HSP90明顯高于SSNS組及正常對照組，治療后SRNS組外周血淋巴細(xì)胞中HSP90較治療前下降(P0.05),但仍高于正常對照組(P0.05)；(4) SRNS組與正常對照組患兒腎組織中P-gp170主要表達(dá)于腎小管及腎間質(zhì)，SRNS組腎組織近曲小管P-gp170陽性率表達(dá)明顯高于正常組(P0.05)，SRNS組內(nèi)近曲小管P-gp170陽性率表達(dá)明顯高于遠(yuǎn)曲小管(P0.05)。(5)正常對照組與SRNS組患兒腎組織中均有HSP90的表達(dá)，多在腎小球及腎小管的胞漿和胞核內(nèi)表達(dá)，SRNS組患兒腎組織中HSP90表達(dá)明顯高于正常對照組(P0.05)。(6)PNS患兒外周血淋巴細(xì)胞中P-gp170及HSP90的表達(dá)分別與24小時(shí)尿蛋白定量正相關(guān)(r分別為0.563,0.785，均P0.05), SRNS組患兒外周血淋巴細(xì)胞中P-gp170表達(dá)與SRNS組患兒腎臟近曲小管中P-gp170的灰度值及病理積分呈等級正相關(guān)(r分別為0.923,0.846，均P0.05), SRNS組患兒外周血淋巴細(xì)胞中HSP90表達(dá)與SRNS組患兒腎臟遠(yuǎn)曲小管中HSP90的灰度值及病理積分呈等級正相關(guān)(r分別為0.708,0.726，均P0.05), SRNS組腎臟病理積分與其24小時(shí)尿蛋白定量高度等級正相關(guān)(r=0.753, P0.05)。 結(jié)論(1)應(yīng)用激素可能誘導(dǎo)PNS患兒外周血淋巴細(xì)胞P-gp170過度表達(dá);(2)SRNS患兒腎組織中P-gp170的表達(dá)強(qiáng)度與病理積分明顯正相關(guān)，外周血淋巴細(xì)胞及腎組織中P-gp170高表達(dá)可能參與了SRNS患兒激素耐藥的發(fā)生和發(fā)展;(3)SRNS患兒外周血淋巴細(xì)胞及腎組織中HSP90高表達(dá)可能與激素耐藥有關(guān);(4)SRNS患兒外周血P-gp170及HSP90的表達(dá)一定程度上可反映腎臟病變的程度。
[Abstract]:Objective to detect the expression of P-glycoprotein 170 (P-gp170) and heat shock protein 90 (HSP90) in peripheral blood lymphocytes and renal tissues of children with primary nephrotic syndrome (PNS) at different time points. To explore the possible mechanism of glucocorticoid (GC) resistance mediated by P-gp170 and HSP90 and its clinical significance. Methods Twenty patients with PNS were enrolled in the affiliated Children's Hospital of Suzhou University. According to the curative effect of GC on children, they were divided into two groups: 10 healthy children with steroid-resistant nephrotic syndrome (SRNS) and 10 steroid-sensitive nephrotic syndrome (SSNS),) as normal control group. Peripheral blood samples were collected at two time points: (1) when GC was not used at the beginning of the disease; (2) after 4 weeks of full GC therapy. Detection of P-gp170 and HSP90 expression in Peripheral Blood lymphocytes of Children with PNS and normal controls by flow Cytometry as follows: 1: 1 flow Cytometry. The expression of P-gp170 and HSP90 in peripheral blood lymphocytes of children with SRNS at different time points. 5 cases of renal biopsy were performed after 4 weeks of hormone therapy. Normal renal tissue as control, The expression of P-gp170 and HSP90 in renal tissue was detected by immunohistochemistry. Results (1) the expression of P-gp170 and HSP90 in peripheral blood lymphocytes of SSNS group and normal control group was higher than that of normal control group (P0.05), (2) the expression of P-gp170 and HSP90 in peripheral blood lymphocytes of SSNS group was higher than that of normal control group before hormone therapy (P0.05). After 4 weeks of hormone therapy, the P-gp170 in peripheral blood lymphocytes in SSNS group and SRNS group was significantly higher than that before treatment (P0.05), and the P-gp170 in peripheral blood lymphocytes of SRNS group was significantly higher than that of SSNS group and normal control group before hormone therapy (P0.05). The level of P-gp170 in peripheral blood lymphocytes in SRNS group was higher than that before treatment (P0.05), and the HSP90 in peripheral blood lymphocytes in SRNS group was significantly higher than that in SSNS group and normal control group before hormone therapy. The level of HSP90 in peripheral blood lymphocytes of SRNS group was lower than that before treatment (P0.05), but still higher than that of normal control group (P0.05). (4) the positive rate of P-gp170 in renal tissue of SRNS group and normal control group was significantly higher than that of normal group (P0.05). The positive rate of P-gp170 in proximal convoluted tubules in renal tubules and interstitial tubules in SRNs group was significantly higher than that in normal control group (P0.05). The positive rate of P-gp170 expression in proximal convoluted tubules was significantly higher than that in distal tubules. The expression of HSP90 was found in renal tissues of normal control group and SRNS group (P0.05). (5). The expression of HSP90 in the renal tissues in the SRNs group was significantly higher than that in the normal control group (P0.05). (6). The expressions of P-gp170 and HSP90 in peripheral blood lymphocytes of the children with PNS were positively correlated with the 24 hour urine protein (r). The expression of P-gp170 in peripheral blood lymphocytes of children in), SRNS group was positively correlated with the gray value and pathological score of P-gp170 in proximal convoluted tubules of SRNS group (r = 0.923 鹵0.846, respectively, P 0.05). The expression of P-gp170 in peripheral blood lymphocytes of children in), SRNS group was positively correlated with the HSP90 table in peripheral blood lymphocytes of children with SRNS group (r = 0.923 鹵0.846, respectively). The grayscale value and pathological score of HSP90 in distal convoluted tubules were positively correlated with those in SRNS group (r = 0.708 鹵0.726, P 0.05). The renal pathological scores in), SRNS group were positively correlated with the 24 hour urinary protein quantitative high grade (r 0.753, P 0.05). Conclusion (1) hormone may induce the overexpression of P-gp170 in peripheral blood lymphocytes of children with PNS, (2) the expression of P-gp170 in renal tissue of children with SRNS is positively correlated with the pathological score. The high expression of P-gp170 in peripheral blood lymphocytes and renal tissues may be involved in the occurrence and development of hormone resistance in children with SRNS; (3) the high expression of HSP90 in peripheral blood lymphocytes and renal tissues of children with SRNS may be related to hormone resistance; (4) the high expression of HSP90 in peripheral blood of SRNS children and children with SRNS; The expression of HSP90 can reflect the degree of renal lesion to some extent.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R726.9

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